@551656AS34@ Group 1 267|Demo User 212 25|1xPLCp3' 27|0 222|3 33|3170 236|484751834 237|484751834 26|762 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL3 53|1 55|3163 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attR2 53|1 55|2508 56|2631 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONRP2R-P3 in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|4 52|PH domain PLCd1 (Rat) 53|0 55|2634 56|3158 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctatggactcgggccgggacttcctgaccctgcacgggctccaggatgacccggaccttcaggcccttctgaagggcagccagcttctgaaggtgaagtccagctcgtggcgtagggaacgcttctacaagctacaggaggactgcaagaccatctggcaggaatctcg 209|aaaggtcatgaggtccccggagtcgcagctgttctccatcgaggacattcaggaggtacggatgggacaccgcacagaaggcctggagaagtttgcccgagacatccccgaggatcgatgcttctccattgtcttcaaggaccagcgcaacaccctagacctcattgccccatcaccagctgacgctcagcactgggtgcagggcctgcgcaagatcatccaccactccggctccatggaccagcggcagaagctgcagcactggattcactcctgcttgcgaaaggctgataaaaacaaggacaacaagatgaacttcaaggagctgaaggacttcctgaaggagctcaacatccagtaggcaacttta 210 212 25|2xAtPH1p3' 27|0 222|3 33|3320 236|484831598 237|485013832 26|764 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL3 53|1 55|3313 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attR2 53|1 55|2508 56|2631 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONRP2R-P3 in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|4 52|PH domain AtPH1 53|0 55|2991 56|3308 57|0 281|1 282|1 283|1 284|1 50 45 51|4 52|PH domain AtPH1 53|0 55|2635 56|2985 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|KGAGAGAGS linker 53|0 55|2958 56|2984 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctctcgagttctggtcaaaccctgagcgttctggttggctcacaaagcaaggcgattacatcaaaacctggcgtcgtcgttggttcgttctcaaacgagggaagcttctctggttcaaagatcaagccgctgctggaattcgtggatctacgccgcgtggtgtgatctc 209|cgttggtgattgtctcaccgtgaaaggagctgaggatgttgtgaataagccttttgcttttgagctatctagtggtagctataccatgttcttcattgctgataatgagaaggagaaagaagagtggattaattcgattggaagatcgattgtgcaaaagggtgctggtgctggtgctggttctctcgagttctggtcaaaccctgagcgttctggttggctcacaaagcaaggcgattacatcaaaacctggcgtcgtcgttggttcgttctcaaacgagggaagcttctctggttcaaagatcaagccgctgctggaattcgtggatctacgccgcgtggtgtgatctccgttggtgattgtctcaccgtgaaaggagctgaggatgttgtgaataagccttttgcttttgagctatctagtggtagctataccatgttcttcattgctgataatgagaaggagaaagaagagtggattaattcgattggaagatcgattgtgcaataggcaacttta 210 212 25|2xCyPET-Hellfire 27|1 222|3 33|1701 236|574343567 237|581540872 26|1669 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|cyPET 53|0 55|52 56|792 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|0 55|790 56|1530 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|Hellfire 53|0 55|1531 56|1698 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|0 55|1561 56|1578 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|3xFlag 53|0 55|1633 56|1698 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|0 55|1606 56|1626 57|0 281|1 282|1 283|1 284|1 50 45 51|4 52|2xCyPet-HellFire 53|0 55|1 56|1698 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|0 55|43 56|72 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|0 55|790 56|810 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|0 55|1534 56|1548 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|atgctcgaggaattcggtaccccgggttcgaaatcgataagcttggatcctctagatgctgctgctgccgctgtgagcaagggagaggaactgttcggcggcatcgtgcccatcctggtggagctggagggcgacgtgaacggccacaagttcagcgtgagcggcgagggcgagggcgacgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgacctggggcgtgcagtgcttcagccggtaccccgaccacatgaagcagcacgacttcttcaagagcgtgatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacatcagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaaggcccggcacaacatcaccgacggcagcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgatcctgcccgacaaccactacctgagcacccagagcgccctgagcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcgtgaccgccgccggcatcacccacggcatggacgaactgtacaaaggactagatgctgctgctgccgctgtgagcaagggagaggaactgttcggcggcatcgtgcccatcctggtggagctggagggcgacgtgaacggccacaagttcagcgtgagcggcgagggcgagggcgacgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgacctggggcgtgcagtgcttcagccggtaccccgaccacatgaagcagcacgacttcttcaagagcgtgatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacatcagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaaggcccggcacaacatcaccgacggcagcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgatcctgcccga 209|caaccactacctgagcacccagagcgccctgagcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcgtgaccgccgccggcatcacccacggcatggacgaactgtacaaaggactagaagctgctgctgctgctggaagaggatcgcatcaccatcaccatcacgattatgatattccaactactgctagcgagaatttgtattttcagggtgagctcgactacaaagaccatgacggtgattataaagatcatgacatcgactacaaggatgacgatgacaagtag 210 212 25|2xFAPP1p3' 27|0 222|3 33|3290 236|481668397 237|481668397 26|721 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL3 53|1 55|3283 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attR2 53|1 55|2508 56|2631 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONRP2R-P3 in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|21 52|PH domain FAPP1 53|0 55|2976 56|3278 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain FAPP1 53|0 55|2640 56|2942 57|0 281|1 282|1 283|1 284|1 50 45 51|15 52|KGAGAGAGS linker 53|0 55|2943 56|2969 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctctcgagatggagggggtgttgtacaagtggaccaactatctcacaggctggcagcctcgttggtttgttttagataatggaatcttatcctactatgattcacaagatgatgtttgcaaagggagcaaaggaagcataaagatggcagtttgtgaaattaaagttca 209|ttcagcagacaacacaagaatggaattaatcattcctggagagcagcatttctacatgaaggcagtgaatgcagctgaaagacagaggtggctggtcgctctggggagctccaaagcatgtttgactgatacaaggactaaaaagggtgctggtgctggtgctggttctctcgagatggagggggtgttgtacaagtggaccaactatctcacaggctggcagcctcgttggtttgttttagataatggaatcttatcctactatgattcacaagatgatgtttgcaaagggagcaaaggaagcataaagatggcagtttgtgaaattaaagttcattcagcagacaacacaagaatggaattaatcattcctggagagcagcatttctacatgaaggcagtgaatgcagctgaaagacagaggtggctggtcgctctggggagctccaaagcatgtttgactgatacaaggactaaataggcaacttta 210 212 25|2xFYVE 27|1 222|3 33|477 236|480527730 237|480529753 26|693 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|FYVE 53|0 55|1 56|231 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|FYVE 53|0 55|247 56|477 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|gaaagtgatgccatgttcgctgctgaaagagcccctgactgggtggatgctgaggaatgccatcggtgcagagtacagtttggggtggtgacccgcaagcatcactgccgagcatgtgggcagatcttctgtggcaagtgctcctccaagtactccaccatccccaagttcggcattgagaaggaggtgcgcgtgtgtgagccctgctatgagcagctgaacaagaaggcacaaggacaaggatccgaaagtgatgccatgttcgctgctgaaagagcccctgactgggtggatgctgaggaatgccatcggtgcagagtacagtttggggtggtgacccgcaagcatcactgccgagcatgtgggcagatcttctgtggcaagtgctcctccaagtactccaccatccccaagttcggcattgagaaggaggtgcgcgtgtgtgagccctgctatgagcagctgaacaagaaggca 210 212 25|2XFYVE (+STOP)/pDONR207 278|GenBank 27|0 222|3 33|3852 236|381169538 237|479122669 26|603 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL1-attL2. 255 224|Dr Peter ROGOWSKY 50 256 224|Dr Peter ROGOWSKY 50 1001|0 1006|27-JUL-2005 45 51|21 52|attL1 53|0 55|3262 56|3357 57|0 281|1 282|1 283|1 284|1 285|3262..3357 50 45 51|21 52|attL2 53|1 55|3846 56|89 57|0 281|1 282|1 283|1 284|1 285|complement(3846..89) 50 45 51|4 52|Gm(R) 53|1 55|873 56|1406 57|0 281|1 282|1 283|1 284|1 285|complement(873..1406) 286|/invitrogen="1150000" 50 45 51|29 52|Pc promoter 53|1 55|1595 56|1622 57|0 281|1 282|1 283|1 284|1 285|complement(1595..1622) 286|/invitrogen="2250000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|3162 56|3205 57|0 281|1 282|1 283|1 284|1 285|3162..3205 286|/invitrogen="2580000" 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|3003 56|3030 57|0 281|1 282|1 283|1 284|1 285|3003..3030 286|/invitrogen="2590000" 50 45 51|27 52|pDONR™207 forward primer 53|0 55|3230 56|3254 57|0 281|1 282|1 283|1 284|1 285|3230..3254 286|/invitrogen="1993000" 50 45 51|27 52|pDONR™207 reverse primer 53|1 55|114 56|137 57|0 281|1 282|1 283|1 284|1 285|complement(114..137) 286|/invitrogen="1994000" 50 45 51|6 52|FYVE 53|0 55|3363 56|3593 57|0 281|1 282|1 283|1 284|1 285|3363..3593 50 45 51|6 52|FYVE 53|0 55|3609 56|3839 57|0 281|1 282|1 283|1 284|1 285|3609..3839 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|cttgtacaaagtgggcattataagaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttgccatccagctgcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaggccgggaagccgatctcggcttgaacgaattgttaggtggcggtacttgggtcgatatcaaagtgcatcacttcttcccgtatgcccaactttgtatagagagccactgcgggatcgtcaccgtaatctgcttgcacgtagatcacataagcaccaagcgcgttggcctcatgcttgaggagattgatgagcgcggtggcaatgccctgcctccggtgctcgccggagactgcgagatcatagatatagatctcactacgcggctgctcaaacctgggcagaacgtaagccgcgagagcgccaacaaccgcttcttggtcgaaggcagcaagcgcgatgaatgtcttactacggagcaagttcccgaggtaatcggagtccggctgatgttgggagtaggtggctacgtctccgaactcacgaccgaaaagatcaagagcagcccgcatggatttgacttggtcagggccgagcctacatgtgcgaatgatgcccatacttgagccacctaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctgcg 209|taacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtacaaaaaaacagtcataacaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgagcgcatacgctacttgcattacagtttacgaaccgaacaggcttatgtcaactgggttcgtgccttcatccgtttccacggtgtgcgtcacccggcaaccttgggcagcagcgaagtcgaggcatttctgtcctggctggcgaacgagcgcaaggtttcggtctccacgcatcgtcaggcattggcggccttgctgttcttctacggcaaggtgctgtgcacggatctgccctggcttcaggagatcggaagacctcggccgtcgcggcgcttgccggtggtgctgaccccggatgaagtggttcgcatcctcggttttctggaaggcgagcatcgtttgttcgcccaggactctagctatagttctagtggttggctactaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcg 209|ggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatctcgggccccaaataatgattttattttgactgatagtgacctgttcgttgcaacaaattgatgagcaatgcttttttataatgccaagtttgtacaaaaaagcaggcttagaaagtgatgccatgttcgctgctgaaagagcccctgactgggtggatgctgaggaatgccatcggtgcagagtacagtttggggtggtgacccgcaagcatcactgccgagcatgtgggcagatcttctgtggcaagtgctcctccaagtactccaccatccccaagttcggcattgagaaggaggtgcgcgtgtgtgagccctgctatgagcagctgaacaagaaggcacaaggacaaggatccgaaagtgatgccatgttcgctgctgaaagagcccctgactgggtggatgctgaggaatgccatcggtgcagagtacagtttggggtggtgacccgcaagcatcactgccgagcatgtgggcagatcttctgtggcaagtgctcctccaagtactccaccatccccaagttcggcattgagaaggaggtgcgcgtgtgtgagccctgctatgagcagctgaacaagaaggcatagacccagcttt 210 212 25|2xGRAMp3' 27|0 222|3 33|3086 236|481713149 237|481713149 26|726 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL3 53|1 55|3079 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attR2 53|1 55|2508 56|2631 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONRP2R-P3 in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|62 52|Y57P 53|0 54|Y57P when the conserved Tyr57 is mutated into a Proline, the GRAM domain lose its ability to bind PI4P 55|3042 56|3044 57|0 281|1 282|1 283|1 284|1 50 45 51|4 52|GRAM domain 53|0 55|2874 56|3074 57|0 281|1 282|1 283|1 284|1 50 45 51|4 52|GRAM domain 53|0 55|2640 56|2840 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|KGAGAGAGS linker 53|0 55|2841 56|2867 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctctcgagtccagatttcgaaagcatttttcccttcctgattcagaggagctactggctagttacttctgccattttcaaaaaaatatacctgtatacggcaaagtatacctaggaacaacttgcatttgctacagatctttatttcccggcacaaacacaactatgat 209|acttccatactccgatattgagaatgtttataatttgaagaagggtgctggtgctggtgctggttctctcgagtccagatttcgaaagcatttttcccttcctgattcagaggagctactggctagttacttctgccattttcaaaaaaatatacctgtatacggcaaagtatacctaggaacaacttgcatttgctacagatctttatttcccggcacaaacacaactatgatacttccatactccgatattgagaatgtttataatttgaagtaggcaacttta 210 212 25|2xOSBPp3' 27|0 222|3 33|3296 236|481668173 237|481668173 26|720 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL3 53|1 55|3289 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attR2 53|1 55|2508 56|2631 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONRP2R-P3 in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|21 52|PH domain OSBP human 53|0 55|2979 56|3284 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain OSBP human 53|0 55|2640 56|2945 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|KGAGAGAGS linker 53|0 55|2946 56|2972 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctagatcttcgggctcggctcgagagggctggctcttcaaatggaccaattatatcaaaggctaccagcggcgatggttcgtgctgagcaacgggctcctgagctactacagatcaaaggcagaaatgagacatacctgccgtggtaccatcaacctcgccacagccaa 209|catcaccgtggaggactcctgcaacttcatcatttccaatgggggtgctcagacctaccatctgaaagctagttcagaagttgagcggcagcgctgggtgacggccctggaactggccaaggccaaagctgtgaagatgctggcaaagggtgctggtgctggtgctggttctagatcttcgggctcggctcgagagggctggctcttcaaatggaccaattatatcaaaggctaccagcggcgatggttcgtgctgagcaacgggctcctgagctactacagatcaaaggcagaaatgagacatacctgccgtggtaccatcaacctcgccacagccaacatcaccgtggaggactcctgcaacttcatcatttccaatgggggtgctcagacctaccatctgaaagctagttcagaagttgagcggcagcgctgggtgacggccctggaactggccaaggccaaagctgtgaagatgctggcataggcaacttta 210 212 25|2xp40phox 27|1 222|3 33|918 236|567963215 237|567963319 26|1595 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|PX domain p40phox 53|0 55|472 56|912 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|PX domain p40phox 53|0 55|1 56|441 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|GAGA linker 53|0 55|442 56|465 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|atggctgtggcccagcagctgcgggccgagagtgactttgaacagcttccggatgatgttgccatctcggccaacattgctgacatcgaggagaagagaggcttcaccagccactttgttttcgtcatcgaggtgaagacaaaaggaggatccaagtacctcatctaccgccgctaccgccagttccatgctttgcagagcaagctggaggagcgcttcgggccagacagcaagagcagtgccctggcctgtaccctgcccacactcccagccaaagtctacgtgggtgtgaaacaggagatcgccgagatgcggatacctgccctcaacgcctacatgaagagcctgctcagcctgccggtctgggtgctgatggatgaggacgtccggatcttcttttaccagtcgccctatgactcagagcaggtgccccaggcactcggtgctggtgctggtgctggttctctcgagatggctgtggcccagcagctgcgggccgagagtgactttgaacagcttccggatgatgttgccatctcggccaacattgctgacatcgaggagaagagaggcttcaccagccactttgttttcgtcatcgaggtgaagacaaaaggaggatccaagtacctcatctaccgccgctaccgccagttccatgctttgcagagcaagctggaggagcgcttcgggccagacagcaagagcagtgccctggcctgtaccctgcccacactcccagccaaagtctacgtgggtgtgaaacaggagatcgccgagatgcggatacctgccctcaacgcctacatgaagagcctgctcagcctgccggtctgggtgctgatggatgaggacgtccggatcttcttttaccagtcgccctatgactcagagcaggtgccccaggcactcctcgag 210 212 25|2xPLCd1 Rat PH 27|1 222|3 33|1071 236|485013144 237|485013361 26|768 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|PH domain PLCd1 Rat 53|0 55|1 56|525 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain PLCd1 Rat 53|0 55|557 56|1068 57|0 281|1 282|1 283|1 284|1 50 45 51|84 52|GAGARS linker 53|0 55|526 56|556 57|0 281|1 282|1 283|1 284|1 50 45 51|4 52|2xPH domain PLCd1 53|0 55|1 56|1068 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|atggactcgggccgggacttcctgaccctgcacgggctccaggatgacccggaccttcaggcccttctgaagggcagccagcttctgaaggtgaagtccagctcgtggcgtagggaacgcttctacaagctacaggaggactgcaagaccatctggcaggaatctcgaaaggtcatgaggtccccggagtcgcagctgttctccatcgaggacattcaggaggtacggatgggacaccgcacagaaggcctggagaagtttgcccgagacatccccgaggatcgatgcttctccattgtcttcaaggaccagcgcaacaccctagacctcattgccccatcaccagctgacgctcagcactgggtgcagggcctgcgcaagatcatccaccactccggctccatggaccagcggcagaagctgcagcactggattcactcctgcttgcgaaaggctgataaaaacaaggacaacaagatgaacttcaaggagctgaaggacttcctgaaggagctcaacatccagggtgcaggagctagatccatggactcgggtagggacttcctgaccctgcacgggctccaggatgacccggaccttcaggcccttctgaagggcagccagcttctgaaggtgaagtccagctcgtggcgtagggaacgcttctacaagctacaggaggactgcaagaccatctggcaggaatctcgaaaggtcatgaggtccccggagtcgcagctgttctccatcgaggacattcaggaggtacggatgggacaccgcacagaaggcctggagaagtttgcccgagacatccccgaggatcgatgcttctccattgtcttcaaggaccagcgcaacaccctagacctcattgccccatcaccagctgacgctcagcactgggtgcagggcctgcgcaagatcatccaccactccggctccatggaccagcggcagaagctgcagcactggattcactcctgcttgcgaaaggctgataaaaacaaggacaacaagatgaacttcaaggagctgaaggacttcctgaaggagctcaacatccagtaa 210 212 25|Ent5 27|1 222|3 33|1236 236|567966662 237|567966779 26|1604 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|4 52|Ent5 53|0 55|1 56|1233 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|ENTH domain Ent5p 53|0 55|1 56|507 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|atggactcattatcaaaaaagattcaaaatttaggtatccatgatattagaaatgccgcaaggtttgcccaaaacgtgatagtccaatatgagccttatcagatagatatccgtcgtgctacaaataccgatgcttggggccccacaccgaagcacctcgccaaggttttaaggaacagataccaagtgccgctctacttgatgacagaatacacgttgaaaagattagtcgaccatatcgctaccaggcccaagaacctatacgaaaaggcaagaaaggactacgttaattatgggtccgaatggagggtggtcttaaaatgtctagtagtcattgaattcctattattgaacgtcgacacaggtgacgaactgaatcagattagatcatgtttgctcactcataagcatattttaaccagggagattgcccaattcaaggtaaaattctccaatgatggtaaaatggagattcatgagagaggtattaggaaaaagggagaacttatcttgcaatacttggaagactcacaatttttgaaaaaggaaagagccaagaataagaagaatgccttgaaaattcgacagcaaggagaatcctccatttataatgccaaccagatttcaacgtctgcaagttacgataacatcgacgacgatgaatttgatgcagatgcagatggctttgacagtgaaatggatgctaataatgtgaccaacttcaatgttcctgtggaaacagaagcaaattcgaatactcgcaggcgttctcatatggaagaacaaagaagacaaagaagggagatattaagagagcagatcaaaaataaagaacagcagaggaaaagaaagcaacaacaagatagcatccctgacttaatcgatcttgacgatagtactagcaccacgaataatataacaatagacaatggtaataatgacaacaaaaacaataatatcaacagcaatagtgatgatgatgatgatgagtttggcgacttccaaagcgaaacgagtcccgacacaacagcacctaaaacttccaatagtaaaatagatgatttgcttgactgggatggcccaaagtcagacacagataccaccgctgctgcgcaaacatcattgccctttgcggagaaaaaacagcaaaaagcccgtcctcaggctacaaaggacaagtcgaagggaaatgatgcattttccgacttattctcttactccaagtcgttggtctaa 210 212 25|GRAM domain (Pichia pastoris) 27|1 222|3 33|246 236|479758257 237|481712880 26|674 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|4 52|GRAM domain 53|0 55|10 56|210 57|0 281|1 282|1 283|1 284|1 50 45 51|62 52|Y57P 53|0 54|Y57P when the conserved Tyr57 is mutated into a Proline, the GRAM domain lose its ability to bind PI4P 55|178 56|180 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|KGAGAGAGS linker 53|0 55|211 56|237 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|aaactcgagtccagatttcgaaagcatttttcccttcctgattcagaggagctactggctagttacttctgccattttcaaaaaaatatacctgtatacggcaaagtatacctaggaacaacttgcatttgctacagatctttatttcccggcacaaacacaactatgatacttccatactccgatattgagaatgtttataatttgaagaagggtgctggtgctggtgctggttctctcgagaaa 210 212 25|mCITRINE 27|1 222|3 33|720 236|473180321 237|473180343 26|452 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|4 52|mCITRINE 53|0 55|1 56|717 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|atggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccttcggctacggcctgatgtgcttcgcccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccaagctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtaa 210 212 25|pDONR221 278|GenBank 27|0 222|1 33|4762 236|38836800 237|312647567 26|380 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Invitrogen 225|(800)-357-3114 226|(760)-602-6500 227|bioinfosales@invitrogen.com 230|Invitrogen Corporation 231|1620 Faraday Avenue 232|Carlsbad, CA 92008 U.S.A. 50 256 224|Invitrogen 225|(800)-357-3114 226|(760)-602-6500 227|bioinfosales@invitrogen.com 230|Invitrogen Corporation 231|1620 Faraday Avenue 232|Carlsbad, CA 92008 U.S.A. 50 42|Plasmid 43|Bacteria 1001|0 1006|25-MAR-2003 45 51|98 52|Invitrogen vector 722 53|0 55|1 56|4762 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="722" 50 45 51|4 52|ccdB 53|1 55|1197 56|1502 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1060000" 50 45 51|4 52|Cm(R) 53|1 55|1847 56|2506 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1080000" 50 45 51|4 52|Kan(R) 53|0 55|3156 56|3965 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|537 56|552 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|517 56|533 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 reverse primer 53|1 55|3027 56|3043 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|T7 primer 53|1 55|3003 56|3022 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|30 52|T7 promoter 53|1 55|3006 56|3022 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|427 56|470 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|268 56|295 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|86 52|attP1 53|0 55|570 56|801 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2685000" 50 45 51|86 52|attP2 53|1 55|2754 56|2985 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2686000" 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccccaaataatgattttattttgactgatagtgacctgttcgttgcaacacattgatgagcaatgcttttttataatgccaactttgtacaaaaaagctgaacgagaaacgtaaaatgatataaatatcaatatattaaattagattttgcataaaaaacagactacataatactgtaaaacacaacatatccagtcactatgaatcaactacttagatggtattagtgacctgtagtcgaccgacagccttccaaatgttcttcgggtgatgctgccaacttagtcgaccgacagccttccaaatgttcttctcaaacggaatcgtcgtatccagcctactcgctattgtcctcaatgccgtattaaatcataaaaagaaataagaaaaagaggtgcgagcctcttttttgtgtgacaaaataaaaacatctacctattcatatacgctagtgtcatagtcctgaaaatcatctgcatcaagaacaatttcacaactcttatacttttctcttacaagtcgttcggcttcatctggattttcagcctctatacttactaaacgtgataaagtttctgtaatttctactgtatcgacctgcagactggctgtgtataagggagcctgacatttatattccccagaacatcaggttaatggcgtttttgatgtcattttcgcggtggctgagatcagccacttcttccccgataacggagaccggcacactggccatatcggtggtcatcatgcgccagctttcatccccgatatgcaccaccgggtaaagttcacgggagactttatctgacagcagacgtgcactggccagggg 209|gatcaccatccgtcgcccgggcgtgtcaataatatcactctgtacatccacaaacagacgataacggctctctcttttataggtgtaaaccttaaactgcatttcaccagcccctgttctcgtcagcaaaagagccgttcatttcaataaaccgggcgacctcagccatcccttcctgattttccgctttccagcgttcggcacgcagacgacgggcttcattctgcatggttgtgcttaccagaccggagatattgacatcatatatgccttgagcaactgatagctgtcgctgtcaactgtcactgtaatacgctgcttcatagcatacctctttttgacatacttcgggtatacatatcagtatatattcttataccgcaaaaatcagcgcgcaaatacgcatactgttatctggcttttagtaagccggatccacgcggcgtttacgccccgccctgccactcatcgcagtactgttgtaattcattaagcattctgccgacatggaagccatcacagacggcatgatgaacctgaatcgccagcggcatcagcaccttgtcgccttgcgtataatatttgcccatggtgaaaacgggggcgaagaagttgtccatattggccacgtttaaatcaaaactggtgaaactcacccagggattggctgagacgaaaaacatattctcaataaaccctttagggaaataggccaggttttcaccgtaacacgccacatcttgcgaatatatgtgtagaaactgccggaaatcgtcgtggtattcactccagagcgatgaaaacgtttcagtttgctcatggaaaacggtgtaacaagggtgaacactatcccatatcaccagctcaccgtctttcattgccatacggaattccggatgagcattcatcaggcgggcaagaatgtgaataaaggccggataaaacttgtgcttatttttctttacggtctttaaaaaggccgtaatatccagctgaacggtctggttataggtacattgagcaactgactgaaatgcctcaaaatgttctttacgatgccattgggatatatcaacggtggtatatccagtgatttttttctccattttagcttccttagctcctgaaaatctcgataactcaaaaaatacgcccggtagtgatcttatttcattatggtgaaagttggaacctcttacgtgccgatcaacgtctcattttcgccaaaagttggcccagggcttcccggtatcaacagggacaccaggatttatttattctgcgaagtgatcttccgtcacaggtatttattcggcgcaaagtgcgtcgggtgatgctgccaacttagtcgactacaggtcactaataccatctaagtagttgattcatagtgactggat 209|atgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcagctttcttgtacaaagttggcattataagaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttgccatccagctgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaataatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagc 209|taccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgtt 210 212 25|pDONRP2R-P3 (p3') 278|GenBank 27|0 222|1 33|4773 236|38836800 237|312647567 26|382 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|Complementary copy of _pDONRTP2R-P3. 255 224|Invitrogen 225|(800)-357-3114 226|(760)-602-6500 227|bioinfosales@invitrogen.com 230|Invitrogen Corporation 231|1620 Faraday Avenue 232|Carlsbad, CA 92008 U.S.A. 50 256 224|Invitrogen 225|(800)-357-3114 226|(760)-602-6500 227|bioinfosales@invitrogen.com 230|Invitrogen Corporation 231|1620 Faraday Avenue 232|Carlsbad, CA 92008 U.S.A. 50 1001|0 1006|24-JUL-2002 45 51|98 52|Invitrogen vector 731 53|0 55|1 56|4773 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="731" 50 45 51|4 52|ccdB 53|0 55|2084 56|2389 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1060000" 50 45 51|4 52|Cm(R) 53|0 55|1083 56|1742 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1080000" 50 45 51|4 52|Kan(R) 53|0 55|3167 56|3976 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|537 56|552 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|517 56|533 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 reverse primer 53|1 55|3038 56|3054 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|T7 primer 53|1 55|3014 56|3033 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|30 52|T7 promoter 53|1 55|3017 56|3033 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|427 56|470 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|268 56|295 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|86 52|attP2R 53|0 55|591 56|822 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2686001" 50 45 51|86 52|attP3 53|0 55|2746 56|2977 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2687000" 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctctagagcgtcgactaagttggcagcatcacccgacgcactttgcgccgaataaatacctgtgacggaagatcacttcgcagaataaataaatcctggtgtccctgttgataccgggaagccctgggccaacttttggcgaaaatgagacgttgatcggcacgtaagaggttccaactttcaccataatgaaataagatcactaccgggcgtattttttgagttatcgagattttcaggagctaaggaagctaaaatggagaaaaaaatcactggatataccaccgttgatatatcccaatggcatcgtaaagaacattttgaggcatttcagtcagttgctcaatgtacctataaccagaccgttcagctggatattacggcctttttaaagaccgtaaagaaaaataagcacaagttttatccggcctttattcacattcttgcccgcctgatgaatgctcatccggaattccgtatggcaatgaaagacggtgagctggtgatatgggatagtgttcacccttgttacaccgttttccatgagcaaactgaaacgttttcatcgctctgg 209|agtgaataccacgacgatttccggcagtttctacacatatattcgcaagatgtggcgtgttacggtgaaaacctggcctatttccctaaagggtttattgagaatatgtttttcgtctcagccaatccctgggtgagtttcaccagttttgatttaaacgtggccaatatggacaacttcttcgcccccgttttcaccatgggcaaatattatacgcaaggcgacaaggtgctgatgccgctggcgattcaggttcatcatgccgtctgtgatggcttccatgtcggcagaatgcttaatgaattacaacagtactgcgatgagtggcagggcggggcgtaatcgcgtggatccggcttactaaaagccagataacagtatgcgtatttgcgcgctgatttttgcggtataagaatatatactgatatgtatacccgaagtatgtcaaaaagaggtgtgctatgaagcagcgtattacagtgacagttgacagcgacagctatcagttgctcaaggcatatatgatgtcaatatctccggtctggtaagcacaaccatgcagaatgaagcccgtcgtctgcgtgccgaacgctggaaagcggaaaatcaggaagggatggctgaggtcgcccggtttattgaaatgaacggctcttttgctgacgagaacagggactggtgaaatgcagtttaaggtttacacctataaaagagagagccgttatcgtctgtttgtggatgtacagagtgatattattgacacgcccgggcgacggatggtgatccccctggccagtgcacgtctgctgtcagataaagtctcccgtgaactttacccggtggtgcatatcggggatgaaagctggcgcatgatgaccaccgatatggccagtgtgccggtctccgttatcggggaagaagtggctgatctcagccaccgcgaaaatgacatcaaaaacgccattaacctgatgttctggggaatataaatgtcaggctcccttatacacagccagtctgcaggtcgatacagtagaaattacagaaactttatcacgtttagtaagtatagaggctgaaaatccagatgaagccgaacgacttgtaagagaaaagtataagagttgtgaaattgttcttgatgcagatgattttcaggactatgacactagcgtatatgaataggtagatgtttttattttgtcacacaaaaaagaggctcgcacctctttttcttatttctttttatgatttaatacggcattgaggacaatagcgagtaggctggatacgacgattccgtttgagaagaacatttggaaggctgtcggtcgagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtg 209|ttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttattatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgcc 209|ggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgtt 210 212 25|pDONRP4-P1R (p5') 278|GenBank 27|0 222|1 33|4777 236|38836800 237|312647567 26|383 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|Ligation of ApaI-P4-ccdB-cat-P1R into pDONRT21 backbone. 255 224|Invitrogen 225|(800)-357-3114 226|(760)-602-6500 227|bioinfosales@invitrogen.com 230|Invitrogen Corporation 231|1620 Faraday Avenue 232|Carlsbad, CA 92008 U.S.A. 50 256 224|Invitrogen 225|(800)-357-3114 226|(760)-602-6500 227|bioinfosales@invitrogen.com 230|Invitrogen Corporation 231|1620 Faraday Avenue 232|Carlsbad, CA 92008 U.S.A. 50 1001|0 1006|24-JUL-2002 1008|SYN 45 51|98 52|Invitrogen vector 732 53|0 55|1 56|4777 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="732" 50 45 51|4 52|ccdB 53|1 55|1181 56|1486 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1060000" 50 45 51|4 52|Cm(R) 53|1 55|1828 56|2487 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1080000" 50 45 51|4 52|Kan(R) 53|0 55|3171 56|3980 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|537 56|552 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|517 56|533 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 reverse primer 53|1 55|3042 56|3058 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|T7 primer 53|1 55|3018 56|3037 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|30 52|T7 promoter 53|1 55|3021 56|3037 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|427 56|470 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|268 56|295 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|86 52|attP4 53|1 55|593 56|824 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2688000" 50 45 51|86 52|attP1R 53|1 55|2748 56|2979 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2685001" 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggcccgcgttaacgctaccatggagctccaaataatgattttattttgactgatagtgacctgttcgttgcaacaaattgataagcaatgcttttttataatgccaactttgtatagaaaagttgaacgagaaacgtaaaatgatataaatatcaatatattaaattagattttgcataaaaaacagactacataatactgtaaaacacaacatatgcagtcactatgaatcaactacttagatggtattagtgacctgtagaattcgagctcgaccgacagccttccaaatgttcttctcaaacggaatcgtcgtatccagcctactcgctattgtcctcaatgccgtattaaatcataaaaagaaataagaaaaagaggtgcgagcctcttttttgtgtgacaaaataaaaacatctacctattcatatacgctagtgtcatagtcctgaaaatcatctgcatcaagaacaatttcacaactcttatacttttctcttacaagtcgttcggcttcatctggattttcagcctctatacttactaaacgtgataaagtttctgtaatttctactgtatcgacctgcagactggctgtgtataagggagcctgacatttatattccccagaacatcaggttaatggcgtttttgatgtcattttcgcggtggctgagatcagccacttcttccccgataacggagaccggcacactggccatatcggtggtcatcatgcgccagctttcatccccgatatgcaccaccgggtaaagttcacgggagactttatctgacagcagacgtgcactggccagggggatcaccatccgtcgc 209|ccgggcgtgtcaataatatcactctgtacatccacaaacagacgataacggctctctcttttataggtgtaaaccttaaactgcatttcaccagtccctgttctcgtcagcaaaagagccgttcatttcaataaaccgggcgacctcagccatcccttcctgattttccgctttccagcgttcggcacgcagacgacgggcttcattctgcatggttgtgcttaccagaccggagatattgacatcatatatgccttgagcaactgatagctgtcgctgtcaactgtcactgtaatacgctgcttcatagcacacctctttttgacatacttcgggtatacatatcagtatatattcttataccgcaaaaatcagcgcgcaaatacgcatactgttatctggcttttagtaagccggatccacgcgattacgccccgccctgccactcatcgcagtactgttgtaattcattaagcattctgccgacatggaagccatcacagacggcatgatgaacctgaatcgccagcggcatcagcaccttgtcgccttgcgtataatatttgcccatggtgaaaacgggggcgaagaagttgtccatattggccacgtttaaatcaaaactggtgaaactcacccagggattggctgagacgaaaaacatattctcaataaaccctttagggaaataggccaggttttcaccgtaacacgccacatcttgcgaatatatgtgtagaaactgccggaaatcgtcgtggtattcactccagagcgatgaaaacgtttcagtttgctcatggaaaacggtgtaacaagggtgaacactatcccatatcaccagctcaccgtctttcattgccatacggaattccggatgagcattcatcaggcgggcaagaatgtgaataaaggccggataaaacttgtgcttatttttctttacggtctttaaaaaggccgtaatatccagctgaacggtctggttataggtacattgagcaactgactgaaatgcctcaaaatgttctttacgatgccattgggatatatcaacggtggtatatccagtgatttttttctccattttagcttccttagctcctgaaaatctcgataactcaaaaaatacgcccggtagtgatcttatttcattatggtgaaagttggaacctcttacgtgccgatcaacgtctcattttcgccaaaagttggcccagggcttcccggtatcaacagggacaccaggatttatttattctgcgaagtgatcttccgtcacaggtatttattcggcgcaaagtgcgtcgggtgatgctgccaacttagtcgacgctctagagctccaaataatgattttattttgactgatagtgacctgttcgttgcaacaaattgat 209|aagcaatgcttttttataatgccaactttgtacaaaaaagttgaacgagaaacgtaaaatgatataaatatcaatatattaaattagattttgcataaaaaacagactacataatactgtaaaacacaacatatgcagtcactatgaatcaactacttagatggtattagtgacctgtagaattcgagctctagagctgcagggcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtt 209|tgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgtt 210 212 25|PH domain FAPP1 human 27|1 222|3 33|351 236|481662992 237|481666854 26|713 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|PH domain FAPP1 53|0 55|10 56|312 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|KGAGAGAGS linker 53|0 55|313 56|339 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|aaactcgagatggagggggtgttgtacaagtggaccaactatctcacaggctggcagcctcgttggtttgttttagataatggaatcttatcctactatgattcacaagatgatgtttgcaaagggagcaaaggaagcataaagatggcagtttgtgaaattaaagttcattcagcagacaacacaagaatggaattaatcattcctggagagcagcatttctacatgaaggcagtgaatgcagctgaaagacagaggtggctggtcgctctggggagctccaaagcatgtttgactgatacaaggactaaaaagggtgctggtgctggtgctggttctctcgagaaataa 210 212 25|PH domain OSBP human 27|1 222|3 33|351 236|481667258 237|481667651 26|718 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|PH domain OSBP human 53|0 55|10 56|315 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|KGAGAGAGS linker 53|0 55|316 56|342 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|aaaagatcttcgggctcggctcgagagggctggctcttcaaatggaccaattatatcaaaggctaccagcggcgatggttcgtgctgagcaacgggctcctgagctactacagatcaaaggcagaaatgagacatacctgccgtggtaccatcaacctcgccacagccaacatcaccgtggaggactcctgcaacttcatcatttccaatgggggtgctcagacctaccatctgaaagctagttcagaagttgagcggcagcgctgggtgacggccctggaactggccaaggccaaagctgtgaagatgctggcaaagggtgctggtgctggtgctggttctagatctaaa 210 212 25|PH domain OSH2p 27|1 222|3 33|1026 236|481714578 237|483542551 26|729 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|4 52|PH domain OSH2p 53|0 55|1 56|522 57|0 281|1 282|1 283|1 284|1 50 45 51|4 52|PH domain OSH2p 53|0 55|523 56|1026 57|0 281|1 282|1 283|1 284|1 50 45 51|45 52|VNSKL linker 53|0 55|502 56|522 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ccgtcgaacaacgttacaccggaaattaagttaaagaatctacttgaaaagaatctgagagaacaagctattgttcacgaagatgtcgcatcaagtaagcctcctacgtataaggggtttttaaagaaatggacaaattttgcccatggctacaaattacgttggttcattttaagtggcgacggaaatctgtcatattacaaagaccaatcgcatgttgataggcctcgtggtactttaaaagtttcgacgtgccgcttacacatcgattcttcagaaaaactgaatttcgagttgctaggtggcatcacaggcacaacaaggtggcgcttaaagggcaatcatcccattgaaacgactaggtgggtcaatgctatacaaagcgctattagattcgccaaagacaaggagatattaaacaaaaaaaaggcagtaccaccatcattggccttgaagaacaaatctccagccctgatatcgcattccaaaacacaaggatctctaccggtgaattcgaagcttccgtcgaacaacgttacaccggaaattaagttaaagaatctacttgaaaagaatctgagagaacaagctattgttcacgaagatgtcgcatcaagtaagcctcctacgtataaggggtttttaaagaaatggacaaattttgcccatggctacaaattacgttggttcattttaagtggcgacggaaatctgtcatattacaaagaccaatcgcatgttgataggcctcgtggtactttaaaagtttcgacgtgccgcttacacatcgattcttcagaaaaactgaatttcgagttgctaggtggcatcacaggcacaacaaggtggcgcttaaagggcaatcatcccattgaaacgactaggtgggtcaatgctatacaaagcgctattagattcgccaaagacaaggagatattaaacaaaaaaaaggcagtaccaccatcattggccttgaagaacaaatctccagccctgatatcgcattccaaaacacaaggatctctt 210 212 25|pNIGEL 16 (pGII(A)-KAN-UBQ10:3H-CER_HOST) 278|GenBank 27|0 222|3 33|7577 236|420117958 237|473184180 26|455 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Niko Geldner 50 256 224|Niko Geldner 50 1001|0 1006|30-MAR-2007 45 51|21 52|RB 53|0 54|right T-DNA border 55|3364 56|3416 57|0 281|1 282|1 283|1 284|1 285|3364..3416 50 45 51|33 52|Replication origin pSa 53|0 55|5448 56|5943 57|0 281|1 282|1 283|1 284|1 285|5448..5943 50 45 51|28 52|T7 53|0 55|1 56|20 57|0 281|1 282|1 283|1 284|1 285|1..20 50 45 51|28 52|T3 53|1 55|2973 56|2992 57|0 281|1 282|1 283|1 284|1 285|complement(2973..2992) 50 45 51|4 52|Amp-Resistence 53|1 55|4244 56|5101 57|0 281|1 282|1 283|1 284|1 285|complement(4244..5101) 50 45 51|52 52|5'UTR 53|0 55|1520 56|2019 57|0 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|0 55|1631 56|1715 57|0 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|0 55|1716 56|2019 57|0 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|0 55|34 56|1630 57|0 281|1 282|1 283|1 284|1 285|34..1630 50 45 51|35 52|HA 1 53|0 55|2031 56|2063 57|0 281|1 282|1 283|1 284|1 285|2031..2063 50 45 51|21 52|HA 2 53|0 55|2064 56|2096 57|0 281|1 282|1 283|1 284|1 285|2064..2096 50 45 51|21 52|HA 3 53|0 55|2097 56|2129 57|0 281|1 282|1 283|1 284|1 285|2097..2129 50 45 51|35 52|loxP 53|0 55|2880 56|2913 57|0 281|1 282|1 283|1 284|1 285|2880..2913 50 45 51|4 52|3xHA-CFP 53|0 55|2022 56|2879 57|0 281|1 282|1 283|1 284|1 285|2022..2879 50 45 51|21 52|EcoRV/HpaI 53|0 55|7388 56|7393 57|0 281|1 282|1 283|1 284|1 285|7388..7393 50 45 51|21 52|HpaI/EcoRV 53|0 55|5988 56|5993 57|0 281|1 282|1 283|1 284|1 285|5988..5993 50 45 51|29 52|nos promoter region 53|1 55|7096 56|7387 57|0 281|1 282|1 283|1 284|1 285|complement(7096..7387) 50 45 51|4 52|KAN-Res-ORF 53|1 55|6304 56|7095 57|0 281|1 282|1 283|1 284|1 285|complement(6304..7095) 50 45 51|43 52|nos terminator region 53|1 55|5994 56|6300 57|0 281|1 282|1 283|1 284|1 285|complement(5994..6300) 50 45 51|21 52|LB 53|0 54|Left T-DNA border 55|5959 56|5990 57|0 281|1 282|1 283|1 284|1 285|5959..5990 50 45 51|21 52|S72A Cerulean Mutation 53|0 55|2373 56|2375 57|0 281|1 282|1 283|1 284|1 285|2373..2375 50 45 51|21 52|Y145A Cerulean Mutation 53|0 55|2592 56|2594 57|0 281|1 282|1 283|1 284|1 285|2592..2594 50 45 51|21 52|H148D Cerulean Mutation 53|0 55|2601 56|2603 57|0 281|1 282|1 283|1 284|1 285|2601..2603 50 45 51|21 52|A to K monomeric mutation 53|0 55|2775 56|2777 57|0 281|1 282|1 283|1 284|1 285|2775..2777 50 45 51|21 52|GCA (Ala) instead of GGA (Gly) acc. to 060831 53|1 55|6664 56|6666 57|0 281|1 282|1 283|1 284|1 285|complement(6664..6666) 50 45 51|21 52|CAC (His) instead of CAG (Gln) acc. to 060831 53|1 55|6533 56|6534 57|0 281|1 282|1 283|1 284|1 285|complement(6533..6534) 50 205 37|0 38|0 39|0 40|0 250 248|http://www.informaxinc.com/ 50 1024|-1 24 209|taatacgactcactatagggcgaattgggtaccagtctagctcaacagagcttttaacccaaattggtacaatagaatacaactttagatcataattctcaaaagaaagagattccttagctattctatctgccactccatttccttctcggcttgtatgcacaagcataaaatcctcaaacttgctaagtagatactttatgtcttggataattggattgagacttgacaagcataactttcatgtaaccaaagacacaagttgctgagaatccacctcaaaaatgatcttcctataattgaatcgggataatgacagcacagcccatctaagagcctccacttctacttccagcacgcttcttacttttaccacagctcttgcacctaaccataacaccttccctgtatgatcgcgaagcacccaccctaagccacattttaatccttctgttggccatgccccatcaaagttgcacttaacccaagattgtggtggagcttcccatgtttctcgtctgtcccgacggtgttgtggttggtgctttccttacattctgagcctctttccttctaatccactcatctgcatcttcttgtgtccttactaatacctcattggttccaaattccctccctttaagcaccagctcgtttctgttcttccacagcctcccaagtatccaagggactaaagcctccacattcttcagatcaggatattcttgtttaagatgttgaactctatggaggtttgtatgaactgatgatctaggaccggataagttcccttcttcatagcgaacttattcaaagaatgttttgtgtatcattcttgttacattgttattaatgaaaaaatattattggtcattggactgaacacgagtgttaaatatggaccaggccccaaataagatccattgatatatgaattaaataacaagaataaatcgagtcaccaaaccacttgccttttttaacgagacttgttcaccaacttgatacaaaagtcattatcctatgcaaatcaataatcatacaaaaatatccaataacactaaaaaattaaaagaaatggataatttcacaatatgttatacgataaagaagttacttttccaagaaattcactgattttataagcccacttgcattagataaatggcaaaaaaaaacaaaaaggaaaagaaataaagcacgaagaattctagaaaatacgaaatacgcttcaatgcagtgggacccacggttcaattattgccaattttcagctccaccgtatatttaaaaaataaaacgataatgctaaaaaaatataaatcgtaacgatcgttaaatctcaacggctggatcttatgacgaccgttagaaattgtggttgtcgacgagtcagtaat 209|aaacggcgtcaaagtggttgcagccggcacacacgagtcgtgtttatcaactcaaagcacaaatacttttcctcaacctaaaaataaggcaattagccaaaaacaactttgcgtgtaaacaacgctcaatacacgtgtcattttattattagctattgcttcaccgccttagctttctcgtgacctagtcgtcctcgtcttttcttcttcttcttctataaaacaatacccaaagagctcttcttcttcacaattcagatttcaatttctcaaaatcttaaaaactttctctcaattctctctaccgtgatcaaggtaaatttctgtgttccttattctctcaaaatcttcgattttgttttcgttcgatcccaatttcgtatatgttctttggtttagattctgttaatcttagatcgaagacgattttctgggtttgatcgttagatatcatcttaattctcgattagggtttcatagatatcatccgatttgttcaaataatttgagttttgtcgaataattactcttcgatttgtgatttctatctagatctggtgttagtttctagtttgtgcgatcgaatttgtcgattaatctgagtttttctgattaacagccatggcaggttacccatacgacgttcctgactatgcgtcactctacccctatgacgtaccggattatgcatccctatatccgtatgatgttccagattacgcttctctacgttcctctagaggcgtccaccatatgatggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgacctggggcgtgcagtgcttcgcccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaacgccatcagcgacaacgtctatatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagcacccagtccaagctgagcaaagaccccaacgagaa 209|gcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagcatatgataacttcgtatagcatacattatacgaagttatcccgggggatccactagttctagagcggccgccaccgcggtggagctccagcttttgttccctttagtgagggttaattccgagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacacaacatacgagccggaaghcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgaaggccttgacaggatatattggcgggtaaactaagtcgctgtatgtgtttgtttgagatctcatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaagaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagtttta 209|aatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgatgatatatttttatcttgtgcaatgtaacatcagagattttgagacacaacgtggctttgttgaataaatcgaacttttgctgagttgaaggatcagatcacgcatcttcccgacaacgcagaccgttccgtggcaaagcaaaagttcaaaatcaccaactggtccacctacaacaaagctctcatcaaccgtggctccctcactttctggctggatgatggggcgattcaggcgatccccatccaacagcccgccgtcgagcgggcttttttatccccggaagcctgtggatagagggtagttatccacgtgaaaccgctaatgccccgcaaagccttgattcacggggctttccggcccgctccaaaaactatccacgtgaaatcgctaatcagggtacgtgaaatcgctaatcggagtacgtgaaatcgctaataaggtcacg 209|tgaaatcgctaatcaaaaaggcacgtgagaacgctaatagccctttcagatcaacagcttgcaaacacccctcgctccggcaagtagttacagcaagtagtatgttcaattagcttttcaattatgaatatatatatcaattattggtcgcccttggcttgtggacaatgcgctacgcgcaccggctccgcccgtggacaaccgcaagcggttgcccaccgtcgagcgccagcgcctttgcccacaacccggcggccggccgcaacagatcgttttataaatttttttttttgaaaaagaaaaagcccgaaaggcggcaacctctcgggcttctggatttccgatccccggaattagagatcttggcaggatatattgtggtgtaacgttatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcaaaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacaggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatca 209|gagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatcgataacattaacgtttacaatttcgcgccattcgccattcaggctgcgcaactgttgggaagggcgatcggtgcgggcctcttcgctattacgccagctggcgaaagggggatgtgctgcaaggcgattaagttgggtaacgccagggttttcccagtcacgacgttgtaaaacgacggccagtgaattg 210 212 25|tubby domain 27|1 222|3 33|792 236|567964944 237|567965043 26|1599 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|Tubby domain 53|0 55|1 56|789 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|gtggatattgaggtccaggatctagaggagtttgcactgaggccagccccacaagggatcaccatcaaatgccgcatcactcgggacaagaaggggatggaccgcggcatgtaccccacctactttctgcacctagaccgtgaggatggcaagaaggtgttcctcctggcgggcaggaagagaaagaagagtaaaacttccaattacctcatctctgtggacccaacagacttgtctcggggaggcgatagctatatcgggaaattgcggtccaacctgatgggcaccaagttcaccgtttatgacaatggcgtcaaccctcagaaggcatcctcttccacgctggaaagcggaaccttgcgccaggagctggcagcggtgtgctatgagacaaatgtcctaggcttcaagggacctcggaagatgagtgtgatcgtcccaggcatgaacatggttcatgagagagtctgtatccgcccccgcaatgaacatgagaccctgttagcacgctggcagaacaagaacacggagagcatcattgagctgcagaacaagacgccagtctggaatgatgacacacagtcctatgtacttaacttccacggccgtgtcacacaggcttctgtgaagaacttccagatcatccacggcaatgacccggactacatcgtcatgcagtttggccgggtagcagaagatgtgttcaccatggattacaactacccactgtgtgcactgcaggcctttgccattgctctgtccagctttgacagcaagctggcctgcgagtag 210 212 25|1xEnt5p221 27|0 222|3 33|3057 236|567966818 237|567966818 26|1605 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL1-attL2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL1 53|0 55|2435 56|2530 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attL2 53|1 55|3050 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONR221 in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2133 56|2160 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2292 56|2335 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|109 56|125 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|106 56|125 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|130 56|146 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2382 56|2398 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2402 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|259 56|1068 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|21 52|ENTH domain Ent5p 53|0 55|2539 56|3045 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONR221 72|0 73|0 74|1 75|2517 76|1 77|2517 78|2898 79|652 80|0 84 91|2 92|0 93|2898 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|652 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|tacaaaaaagcaggcttaacc 72|0 73|0 74|1 75|21 76|2518 77|2538 78|1 79|21 80|0 81|tacaaaaaagcaggcttaacc 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|Ent5 72|0 73|0 74|1 75|507 76|2539 77|3045 78|1 79|507 80|0 84 91|2 92|0 93|1 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|507 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|tacccagctttc 72|0 73|0 74|1 75|12 76|3046 77|3057 78|1 79|12 80|0 81|tacccagctttc 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttgtacaaagttggcattataagaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttgccatccagctgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaataatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaa 209|ctctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccccaaataatgattttattttgactgatagtgacctgttcgttgcaacacattgatgagcaatgcttttttataatgccaactttgtacaaaaaagcaggcttaaccatggactcattatcaaaaaagattcaaaatttaggtatccatgatattagaaatgccgcaaggtttgcccaaaacgtgatagtccaatatgagccttatcagatagatatccgtcgtgctacaaataccgatgcttggggccccacaccgaagcacctcgccaaggttttaaggaacagataccaagtgccgctctacttgatgacagaatacacgttgaaaagattagtcgaccatatcgctaccaggcccaagaacctat 209|acgaaaaggcaagaaaggactacgttaattatgggtccgaatggagggtggtcttaaaatgtctagtagtcattgaattcctattattgaacgtcgacacaggtgacgaactgaatcagattagatcatgtttgctcactcataagcatattttaaccagggagattgcccaattcaaggtaaaattctccaatgatggtaaaatggagattcatgagagaggtattaggaaaaagggagaactttacccagctttc 210 212 25|1xFAPP1p3' 27|0 222|3 33|2954 236|481666963 237|481666963 26|716 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|21 52|PH domain FAPP1 53|0 55|2640 56|2942 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|2618 76|1 77|2618 78|2890 79|734 80|0 84 91|2 92|0 93|2890 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|734 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ttgtacaaagtggct 72|0 73|0 74|1 75|15 76|2619 77|2633 78|1 79|15 80|0 81|ttgtacaaagtggct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|PH domain FAPP1 human 72|0 73|0 74|1 75|309 76|2634 77|2942 78|4 79|312 80|0 84 91|2 92|0 93|4 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|312 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|taggcaacttta 72|0 73|0 74|1 75|12 76|2943 77|2954 78|1 79|12 80|0 81|taggcaacttta 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctctcgagatggagggggtgttgtacaagtggaccaactatctcacaggctggcagcctcgttggtttgttttagataatggaatcttatcctactatgattcacaagatgatgtttgcaaagggagcaaaggaagcataaagatggcagtttgtgaaattaaagttca 209|ttcagcagacaacacaagaatggaattaatcattcctggagagcagcatttctacatgaaggcagtgaatgcagctgaaagacagaggtggctggtcgctctggggagctccaaagcatgtttgactgatacaaggactaaataggcaacttta 210 212 25|1xFYVEp3' 27|0 222|3 33|2876 236|480779146 237|480779146 26|699 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|21 52|FYVE 53|0 55|2634 56|2864 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|2618 76|1 77|2618 78|2890 79|734 80|0 84 91|2 92|0 93|2890 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|734 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ttgtacaaagtggct 72|0 73|0 74|1 75|15 76|2619 77|2633 78|1 79|15 80|0 81|ttgtacaaagtggct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|2xFYVE 72|0 73|0 74|1 75|231 76|2634 77|2864 78|1 79|231 80|0 84 91|2 92|0 93|1 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|231 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|taggcaacttta 72|0 73|0 74|1 75|12 76|2865 77|2876 78|1 79|12 80|0 81|taggcaacttta 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctgaaagtgatgccatgttcgctgctgaaagagcccctgactgggtggatgctgaggaatgccatcggtgcagagtacagtttggggtggtgacccgcaagcatcactgccgagcatgtgggcagatcttctgtggcaagtgctcctccaagtactccaccatccccaa 209|gttcggcattgagaaggaggtgcgcgtgtgtgagccctgctatgagcagctgaacaagaaggcataggcaacttta 210 212 25|1xGRAM domain/pDONRP2R-P3 27|0 222|3 33|2846 236|481888255 237|481888255 26|738 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|62 52|Y57P 53|0 54|Y57P when the conserved Tyr57 is mutated into a Proline, the GRAM domain lose its ability to bind PI4P 55|2802 56|2804 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|GRAM domain 53|0 55|2634 56|2834 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|2618 76|1 77|2618 78|2890 79|734 80|0 84 91|2 92|0 93|2890 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|734 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ttgtacaaagtggct 72|0 73|0 74|1 75|15 76|2619 77|2633 78|1 79|15 80|0 81|ttgtacaaagtggct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|GRAM domain (Pichia pastoris) 72|0 73|0 74|1 75|201 76|2634 77|2834 78|10 79|210 80|0 84 91|2 92|0 93|10 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|210 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|taggcaacttta 72|0 73|0 74|1 75|12 76|2835 77|2846 78|1 79|12 80|0 81|taggcaacttta 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggcttccagatttcgaaagcatttttcccttcctgattcagaggagctactggctagttacttctgccattttcaaaaaaatatacctgtatacggcaaagtatacctaggaacaacttgcatttgctacagatctttatttcccggcacaaacacaactatgatacttcc 209|atactccgatattgagaatgtttataatttgaagtaggcaacttta 210 212 25|1xOSBPp3' 27|0 222|3 33|2957 236|481667709 237|481667709 26|719 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|21 52|PH domain OSBP human 53|0 55|2640 56|2945 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|2618 76|1 77|2618 78|2890 79|734 80|0 84 91|2 92|0 93|2890 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|734 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ttgtacaaagtggct 72|0 73|0 74|1 75|15 76|2619 77|2633 78|1 79|15 80|0 81|ttgtacaaagtggct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|PH domain OSBP human 72|0 73|0 74|1 75|312 76|2634 77|2945 78|4 79|315 80|0 84 91|2 92|0 93|4 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|315 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|taggcaacttta 72|0 73|0 74|1 75|12 76|2946 77|2957 78|1 79|12 80|0 81|taggcaacttta 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctagatcttcgggctcggctcgagagggctggctcttcaaatggaccaattatatcaaaggctaccagcggcgatggttcgtgctgagcaacgggctcctgagctactacagatcaaaggcagaaatgagacatacctgccgtggtaccatcaacctcgccacagccaa 209|catcaccgtggaggactcctgcaacttcatcatttccaatgggggtgctcagacctaccatctgaaagctagttcagaagttgagcggcagcgctgggtgacggccctggaactggccaaggccaaagctgtgaagatgctggcataggcaacttta 210 212 25|1xOSH2p3' 27|0 222|3 33|3149 236|481714943 237|655477094 26|731 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|21 52|PH domain OSH2p 53|0 55|2634 56|3137 57|0 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|2618 76|1 77|2618 78|2890 79|734 80|0 84 91|2 92|0 93|2890 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|734 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ttgtacaaagtggct 72|0 73|0 74|1 75|15 76|2619 77|2633 78|1 79|15 80|0 81|ttgtacaaagtggct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|PH domain OSH2p 72|0 73|0 74|1 75|504 76|2634 77|3137 78|1 79|504 80|0 84 91|2 92|0 93|1 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|504 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|taggcaacttta 72|0 73|0 74|1 75|12 76|3138 77|3149 78|1 79|12 80|0 81|taggcaacttta 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctccgtcgaacaacgttacaccggaaattaagttaaagaatctacttgaaaagaatctgagagaacaagctattgttcacgaagatgtcgcatcaagtaagcctcctacgtataaggggtttttaaagaaatggacaaattttgcccatggctacaaattacgttggtt 209|cattttaagtggcgacggaaatctgtcatattacaaagaccaatcgcatgttgataggcctcgtggtactttaaaagtttcgacgtgccgcttacacatcgattcttcagaaaaactgaatttcgagttgctaggtggcatcacaggcacaacaaggtggcgcttaaagggcaatcatcccattgaaacgactaggtgggtcaatgctatacaaagcgctattagattcgccaaagacaaggagatattaaacaaaaaaaaggcagtaccaccatcattggccttgaagaacaaatctccagccctgatatcgcattccaaaacacaaggatctctataggcaacttta 210 212 25|1xp40p221 27|0 222|3 33|3021 236|567963418 237|567963418 26|1597 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL1-attL2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL1 53|0 55|2435 56|2530 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attL2 53|1 55|3014 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONR221 in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2133 56|2160 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2292 56|2335 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|109 56|125 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|106 56|125 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|130 56|146 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2382 56|2398 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2402 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|259 56|1068 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|21 52|GAGA linker 53|0 55|2980 56|3003 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PX domain p40phox 53|0 55|2539 56|2979 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONR221 72|0 73|0 74|1 75|2517 76|1 77|2517 78|2898 79|652 80|0 84 91|2 92|0 93|2898 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|652 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|tacaaaaaagcaggcttaacc 72|0 73|0 74|1 75|21 76|2518 77|2538 78|1 79|21 80|0 81|tacaaaaaagcaggcttaacc 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|2xp40phox 72|0 73|0 74|1 75|471 76|2539 77|3009 78|1 79|471 80|0 84 91|2 92|0 93|1 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|471 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|tacccagctttc 72|0 73|0 74|1 75|12 76|3010 77|3021 78|1 79|12 80|0 81|tacccagctttc 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttgtacaaagttggcattataagaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttgccatccagctgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaataatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaa 209|ctctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccccaaataatgattttattttgactgatagtgacctgttcgttgcaacacattgatgagcaatgcttttttataatgccaactttgtacaaaaaagcaggcttaaccatggctgtggcccagcagctgcgggccgagagtgactttgaacagcttccggatgatgttgccatctcggccaacattgctgacatcgaggagaagagaggcttcaccagccactttgttttcgtcatcgaggtgaagacaaaaggaggatccaagtacctcatctaccgccgctaccgccagttccatgctttgcagagcaagctggaggagcgcttcgggccagacagcaagagcagtgccctggcctgtaccctgccca 209|cactcccagccaaagtctacgtgggtgtgaaacaggagatcgccgagatgcggatacctgccctcaacgcctacatgaagagcctgctcagcctgccggtctgggtgctgatggatgaggacgtccggatcttcttttaccagtcgccctatgactcagagcaggtgccccaggcactcggtgctggtgctggtgctggttctctcgagtacccagctttc 210 212 25|1xTUBBYp3' 27|0 222|3 33|3434 236|567965161 237|567965161 26|1601 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL3 53|1 55|3427 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attR2 53|1 55|2508 56|2631 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONRP2R-P3 (p3') in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|21 52|Tubby domain 53|0 55|2634 56|3422 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|2618 76|1 77|2618 78|2890 79|734 80|0 84 91|2 92|0 93|2890 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|734 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ttgtacaaagtggct 72|0 73|0 74|1 75|15 76|2619 77|2633 78|1 79|15 80|0 81|ttgtacaaagtggct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|tubby domain 72|0 73|0 74|1 75|792 76|2634 77|3425 78|1 79|792 80|0 84 91|2 92|0 93|1 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|792 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|gcaacttta 72|0 73|0 74|1 75|9 76|3426 77|3434 78|1 79|9 80|0 81|gcaacttta 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctgtggatattgaggtccaggatctagaggagtttgcactgaggccagccccacaagggatcaccatcaaatgccgcatcactcgggacaagaaggggatggaccgcggcatgtaccccacctactttctgcacctagaccgtgaggatggcaagaaggtgttcctcct 209|ggcgggcaggaagagaaagaagagtaaaacttccaattacctcatctctgtggacccaacagacttgtctcggggaggcgatagctatatcgggaaattgcggtccaacctgatgggcaccaagttcaccgtttatgacaatggcgtcaaccctcagaaggcatcctcttccacgctggaaagcggaaccttgcgccaggagctggcagcggtgtgctatgagacaaatgtcctaggcttcaagggacctcggaagatgagtgtgatcgtcccaggcatgaacatggttcatgagagagtctgtatccgcccccgcaatgaacatgagaccctgttagcacgctggcagaacaagaacacggagagcatcattgagctgcagaacaagacgccagtctggaatgatgacacacagtcctatgtacttaacttccacggccgtgtcacacaggcttctgtgaagaacttccagatcatccacggcaatgacccggactacatcgtcatgcagtttggccgggtagcagaagatgtgttcaccatggattacaactacccactgtgtgcactgcaggcctttgccattgctctgtccagctttgacagcaagctggcctgcgagtaggcaacttta 210 212 25|2xCyPET-3xFLAG-6xHIS/pDONRP2R-P3 (p3') 27|0 222|3 33|4340 236|574349381 237|574349381 26|1670 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL3 53|1 55|4333 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attR2 53|1 55|2508 56|2631 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONRP2R-P3 (p3') in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|21 52|polyA linker 53|0 55|4167 56|4181 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|0 55|3423 56|3443 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|0 55|2676 56|2705 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|0 55|4239 56|4259 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|3xFlag 53|0 55|4266 56|4331 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|0 55|4194 56|4211 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Hellfire 53|0 55|4164 56|4331 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|0 55|3423 56|4163 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|0 55|2685 56|3425 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|2xCyPet-HellFire 53|0 55|2634 56|4331 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|2618 76|1 77|2618 78|2890 79|734 80|0 84 91|2 92|0 93|2890 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|734 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ttgtacaaagtggct 72|0 73|0 74|1 75|15 76|2619 77|2633 78|1 79|15 80|0 81|ttgtacaaagtggct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|2xCyPET-Hellfire 72|0 73|0 74|1 75|1698 76|2634 77|4331 78|1 79|1698 80|0 84 91|2 92|0 93|1 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|1698 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|gcaacttta 72|0 73|0 74|1 75|9 76|4332 77|4340 78|1 79|9 80|0 81|gcaacttta 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctatgctcgaggaattcggtaccccgggttcgaaatcgataagcttggatcctctagatgctgctgctgccgctgtgagcaagggagaggaactgttcggcggcatcgtgcccatcctggtggagctggagggcgacgtgaacggccacaagttcagcgtgagcggcga 209|gggcgagggcgacgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgacctggggcgtgcagtgcttcagccggtaccccgaccacatgaagcagcacgacttcttcaagagcgtgatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacatcagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaaggcccggcacaacatcaccgacggcagcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgatcctgcccgacaaccactacctgagcacccagagcgccctgagcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcgtgaccgccgccggcatcacccacggcatggacgaactgtacaaaggactagatgctgctgctgccgctgtgagcaagggagaggaactgttcggcggcatcgtgcccatcctggtggagctggagggcgacgtgaacggccacaagttcagcgtgagcggcgagggcgagggcgacgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgacctggggcgtgcagtgcttcagccggtaccccgaccacatgaagcagcacgacttcttcaagagcgtgatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacatcagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaaggcccggcacaacatcaccgacggcagcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgatcctgcccgacaaccactacctgagcacccagagcgccctgagcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcgtgaccgccgccggcatcacccacggcatggacgaactgtacaaaggactagaagctgctgctgctgctggaagaggatcgcatcacc 209|atcaccatcacgattatgatattccaactactgctagcgagaatttgtattttcagggtgagctcgactacaaagaccatgacggtgattataaagatcatgacatcgactacaaggatgacgatgacaaggcaacttta 210 212 25|2xFYVEp3' 27|0 222|3 33|3122 236|479122775 237|479122775 26|604 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL3 53|1 55|3115 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attR2 53|1 55|2508 56|2631 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONRP2R-P3 in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|6 52|FYVE 53|0 55|2880 56|3110 57|1 281|1 282|1 283|1 284|1 285|3609..3839 50 45 51|6 52|FYVE 53|0 55|2634 56|2864 57|1 281|1 282|1 283|1 284|1 285|3363..3593 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|2618 76|1 77|2618 78|2890 79|734 80|0 84 91|2 92|0 93|2890 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|734 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ttgtacaaagtggct 72|0 73|0 74|1 75|15 76|2619 77|2633 78|1 79|15 80|0 81|ttgtacaaagtggct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|2XFYVE (+STOP)/pDONR207 72|0 73|0 74|1 75|477 76|2634 77|3110 78|3363 79|3839 80|0 84 91|2 92|0 93|3363 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|3839 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ctagcaacttta 72|0 73|0 74|1 75|12 76|3111 77|3122 78|1 79|12 80|0 81|ctagcaacttta 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctgaaagtgatgccatgttcgctgctgaaagagcccctgactgggtggatgctgaggaatgccatcggtgcagagtacagtttggggtggtgacccgcaagcatcactgccgagcatgtgggcagatcttctgtggcaagtgctcctccaagtactccaccatccccaa 209|gttcggcattgagaaggaggtgcgcgtgtgtgagccctgctatgagcagctgaacaagaaggcacaaggacaaggatccgaaagtgatgccatgttcgctgctgaaagagcccctgactgggtggatgctgaggaatgccatcggtgcagagtacagtttggggtggtgacccgcaagcatcactgccgagcatgtgggcagatcttctgtggcaagtgctcctccaagtactccaccatccccaagttcggcattgagaaggaggtgcgcgtgtgtgagccctgctatgagcagctgaacaagaaggcactagcaacttta 210 212 25|2xOSH2p3' 27|0 222|3 33|3668 236|481714834 237|483542742 26|730 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|4 52|PH domain OSH2 53|0 55|3156 56|3656 57|0 281|1 282|1 283|1 284|1 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|45 52|VNSKL linker 53|0 55|3135 56|3155 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|PH domain OSH2p 53|0 55|2634 56|3155 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|2618 76|1 77|2618 78|2890 79|734 80|0 84 91|2 92|0 93|2890 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|734 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ttgtacaaagtggct 72|0 73|0 74|1 75|15 76|2619 77|2633 78|1 79|15 80|0 81|ttgtacaaagtggct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|PH domain OSH2p 72|0 73|0 74|1 75|1023 76|2634 77|3656 78|1 79|1023 80|0 84 91|2 92|0 93|1 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|1023 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|taggcaacttta 72|0 73|0 74|1 75|12 76|3657 77|3668 78|1 79|12 80|0 81|taggcaacttta 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctccgtcgaacaacgttacaccggaaattaagttaaagaatctacttgaaaagaatctgagagaacaagctattgttcacgaagatgtcgcatcaagtaagcctcctacgtataaggggtttttaaagaaatggacaaattttgcccatggctacaaattacgttggtt 209|cattttaagtggcgacggaaatctgtcatattacaaagaccaatcgcatgttgataggcctcgtggtactttaaaagtttcgacgtgccgcttacacatcgattcttcagaaaaactgaatttcgagttgctaggtggcatcacaggcacaacaaggtggcgcttaaagggcaatcatcccattgaaacgactaggtgggtcaatgctatacaaagcgctattagattcgccaaagacaaggagatattaaacaaaaaaaaggcagtaccaccatcattggccttgaagaacaaatctccagccctgatatcgcattccaaaacacaaggatctctaccggtgaattcgaagcttccgtcgaacaacgttacaccggaaattaagttaaagaatctacttgaaaagaatctgagagaacaagctattgttcacgaagatgtcgcatcaagtaagcctcctacgtataaggggtttttaaagaaatggacaaattttgcccatggctacaaattacgttggttcattttaagtggcgacggaaatctgtcatattacaaagaccaatcgcatgttgataggcctcgtggtactttaaaagtttcgacgtgccgcttacacatcgattcttcagaaaaactgaatttcgagttgctaggtggcatcacaggcacaacaaggtggcgcttaaagggcaatcatcccattgaaacgactaggtgggtcaatgctatacaaagcgctattagattcgccaaagacaaggagatattaaacaaaaaaaaggcagtaccaccatcattggccttgaagaacaaatctccagccctgatatcgcattccaaaacacaaggatcttaggcaacttta 210 212 25|2xPLCp3' 27|0 222|3 33|3713 236|485013418 237|485013418 26|767 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2152 56|2179 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2311 56|2354 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|128 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|125 56|144 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|149 56|165 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2401 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2421 56|2436 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|278 56|1087 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|84 52|GAGARS linker 53|0 55|3159 56|3189 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain PLCd1 Rat 53|0 55|3190 56|3701 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain PLCd1 Rat 53|0 55|2634 56|3158 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|2xPH domain PLCd1 53|0 55|2634 56|3701 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|2618 76|1 77|2618 78|2890 79|734 80|0 84 91|2 92|0 93|2890 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|734 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|ttgtacaaagtggct 72|0 73|0 74|1 75|15 76|2619 77|2633 78|1 79|15 80|0 81|ttgtacaaagtggct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|2xPLCd1 Rat PH 72|0 73|0 74|1 75|1068 76|2634 77|3701 78|1 79|1068 80|0 84 91|2 92|0 93|1 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|1068 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|taggcaacttta 72|0 73|0 74|1 75|12 76|3702 77|3713 78|1 79|12 80|0 81|taggcaacttta 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttatacaaagttggcattataaaaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttggagctccatggtagcgttaacgcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagt 209|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctatggactcgggccgggacttcctgaccctgcacgggctccaggatgacccggaccttcaggcccttctgaagggcagccagcttctgaaggtgaagtccagctcgtggcgtagggaacgcttctacaagctacaggaggactgcaagaccatctggcaggaatctcg 209|aaaggtcatgaggtccccggagtcgcagctgttctccatcgaggacattcaggaggtacggatgggacaccgcacagaaggcctggagaagtttgcccgagacatccccgaggatcgatgcttctccattgtcttcaaggaccagcgcaacaccctagacctcattgccccatcaccagctgacgctcagcactgggtgcagggcctgcgcaagatcatccaccactccggctccatggaccagcggcagaagctgcagcactggattcactcctgcttgcgaaaggctgataaaaacaaggacaacaagatgaacttcaaggagctgaaggacttcctgaaggagctcaacatccagggtgcaggagctagatccatggactcgggtagggacttcctgaccctgcacgggctccaggatgacccggaccttcaggcccttctgaagggcagccagcttctgaaggtgaagtccagctcgtggcgtagggaacgcttctacaagctacaggaggactgcaagaccatctggcaggaatctcgaaaggtcatgaggtccccggagtcgcagctgttctccatcgaggacattcaggaggtacggatgggacaccgcacagaaggcctggagaagtttgcccgagacatccccgaggatcgatgcttctccattgtcttcaaggaccagcgcaacaccctagacctcattgccccatcaccagctgacgctcagcactgggtgcagggcctgcgcaagatcatccaccactccggctccatggaccagcggcagaagctgcagcactggattcactcctgcttgcgaaaggctgataaaaacaaggacaacaagatgaacttcaaggagctgaaggacttcctgaaggagctcaacatccagtaggcaacttta 210 212 25|mCITRINEnoSTOP/pDONR221 27|0 222|3 33|3267 236|479583510 237|479583510 26|655 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL1-attL2 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL1 53|0 55|2435 56|2530 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attL2 53|1 55|3260 56|88 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONR221 in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2133 56|2160 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2292 56|2335 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|109 56|125 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|106 56|125 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|130 56|146 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2382 56|2398 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2402 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|259 56|1068 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|4 52|mCITRINE 53|0 55|2539 56|3255 57|1 281|1 282|1 283|1 284|1 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONR221 72|0 73|0 74|1 75|2517 76|1 77|2517 78|2898 79|652 80|0 84 91|2 92|0 93|2898 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|652 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|tacaaaaaagcaggcttaacc 72|0 73|0 74|1 75|21 76|2518 77|2538 78|1 79|21 80|0 81|tacaaaaaagcaggcttaacc 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|mCITRINE 72|0 73|0 74|1 75|717 76|2539 77|3255 78|1 79|717 80|0 84 91|2 92|0 93|1 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|717 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|tacccagctttc 72|0 73|0 74|1 75|12 76|3256 77|3267 78|1 79|12 80|0 81|tacccagctttc 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ttgtacaaagttggcattataagaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttgccatccagctgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaataatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaa 209|ctctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccccaaataatgattttattttgactgatagtgacctgttcgttgcaacacattgatgagcaatgcttttttataatgccaactttgtacaaaaaagcaggcttaaccatggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccttcggctacggcctgatgtgcttcgcccgctaccccgaccacatgaagcagcacgacttcttcaagtccg 209|ccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccaagctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtacccagctttc 210 212 25|UBQ10prom/pDONRP4-P1R 27|0 222|3 33|4632 236|473263309 237|473263309 26|484 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL4-attR1 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attL4 53|0 55|2540 56|2635 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attR1 53|0 55|4625 56|116 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Generate PCR product using computed primers and a DNA polymerase (i.e. Platinum Pfx or Platinum Taq). Check quality and purify product by PEG/Mg2+ precipitation and agarose gels. Mix purified PCR product and pDONR Vector pDONRP4-P1R in BP buffer and TE buffers. Thaw, vortex and add BP Clonase enzyme. Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a or TOP10 cells and plate on kanamycin plates. Pick colonies and grow in selection media. 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|2215 56|2242 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2590000" 50 45 51|43 52|rrnB T1 transcription terminator 53|0 55|2374 56|2417 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|191 56|207 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|188 56|207 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2100000" 50 45 51|27 52|M13 reverse primer 53|1 55|212 56|228 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2010000" 50 45 51|27 52|M13 (-40) forward primer 53|0 55|2464 56|2480 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="2000000" 50 45 51|27 52|M13 (-20) forward primer 53|0 55|2484 56|2499 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|341 56|1150 57|1 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|29 52|5' Promoter region 53|0 55|2638 56|4234 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 45 51|15 52|Intron 1.1 53|0 55|4320 56|4623 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|21 52|Exon 1.1 53|0 55|4235 56|4319 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|52 52|5'UTR 53|0 55|4124 56|4623 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pDONRP4-P1R (p5') 72|0 73|0 74|1 75|2622 76|1 77|2622 78|2831 79|675 80|0 84 91|2 92|0 93|2831 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|675 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|tatagaaaagttgct 72|0 73|0 74|1 75|15 76|2623 77|2637 78|1 79|15 80|0 81|tatagaaaagttgct 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|pNIGEL 16 (pGII(A)-KAN-UBQ10:3H-CER_HOST) 72|0 73|0 74|1 75|1986 76|2638 77|4623 78|34 79|2019 80|0 84 91|2 92|0 93|34 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|1 218|1 60|0 61|0 63|0 64|gcaagtttg 72|0 73|0 74|1 75|9 76|4624 77|4632 78|1 79|9 80|0 81|gcaagtttg 84 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|0 92|0 93|0 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|tacaaaaaagttgaacgagaaacgtaaaatgatataaatatcaatatattaaattagattttgcataaaaaacagactacataatactgtaaaacacaacatatgcagtcactatgaatcaactacttagatggtattagtgacctgtagaattcgagctctagagctgcagggcggccgcgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttc 209|cgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggcccgcgttaacgctaccatggagctccaaataatgattttattttgactgatagtgacctgttcgttgcaacaaattgataagcaatgcttttttataatgccaactttgtatagaaaagttgctagtctagctcaacagagcttttaacccaaattggtacaatagaatacaactttagatcataattctcaaaagaaagagattccttagctattctatctgccactccatttccttctcggcttgtatgcacaagcataaaatcctcaaacttgctaagtagata 209|ctttatgtcttggataattggattgagacttgacaagcataactttcatgtaaccaaagacacaagttgctgagaatccacctcaaaaatgatcttcctataattgaatcgggataatgacagcacagcccatctaagagcctccacttctacttccagcacgcttcttacttttaccacagctcttgcacctaaccataacaccttccctgtatgatcgcgaagcacccaccctaagccacattttaatccttctgttggccatgccccatcaaagttgcacttaacccaagattgtggtggagcttcccatgtttctcgtctgtcccgacggtgttgtggttggtgctttccttacattctgagcctctttccttctaatccactcatctgcatcttcttgtgtccttactaatacctcattggttccaaattccctccctttaagcaccagctcgtttctgttcttccacagcctcccaagtatccaagggactaaagcctccacattcttcagatcaggatattcttgtttaagatgttgaactctatggaggtttgtatgaactgatgatctaggaccggataagttcccttcttcatagcgaacttattcaaagaatgttttgtgtatcattcttgttacattgttattaatgaaaaaatattattggtcattggactgaacacgagtgttaaatatggaccaggccccaaataagatccattgatatatgaattaaataacaagaataaatcgagtcaccaaaccacttgccttttttaacgagacttgttcaccaacttgatacaaaagtcattatcctatgcaaatcaataatcatacaaaaatatccaataacactaaaaaattaaaagaaatggataatttcacaatatgttatacgataaagaagttacttttccaagaaattcactgattttataagcccacttgcattagataaatggcaaaaaaaaacaaaaaggaaaagaaataaagcacgaagaattctagaaaatacgaaatacgcttcaatgcagtgggacccacggttcaattattgccaattttcagctccaccgtatatttaaaaaataaaacgataatgctaaaaaaatataaatcgtaacgatcgttaaatctcaacggctggatcttatgacgaccgttagaaattgtggttgtcgacgagtcagtaataaacggcgtcaaagtggttgcagccggcacacacgagtcgtgtttatcaactcaaagcacaaatacttttcctcaacctaaaaataaggcaattagccaaaaacaactttgcgtgtaaacaacgctcaatacacgtgtcattttattattagctattgcttcaccgccttagctttctcgtgacctagtcgtcctc 209|gtcttttcttcttcttcttctataaaacaatacccaaagagctcttcttcttcacaattcagatttcaatttctcaaaatcttaaaaactttctctcaattctctctaccgtgatcaaggtaaatttctgtgttccttattctctcaaaatcttcgattttgttttcgttcgatcccaatttcgtatatgttctttggtttagattctgttaatcttagatcgaagacgattttctgggtttgatcgttagatatcatcttaattctcgattagggtttcatagatatcatccgatttgttcaaataatttgagttttgtcgaataattactcttcgatttgtgatttctatctagatctggtgttagtttctagtttgtgcgatcgaatttgtcgattaatctgagtttttctgattaacaggcaagtttg 210 212 25|P3C [UBQ10prom::1xPX(p40)-2xCyPet/pK7m34GW] 27|0 222|3 33|12751 236|655480911 237|655480911 26|2763 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10734 56|10754 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|10230 56|10250 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12744 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UBQ10prom/pDONRP4-P1R, 1xp40p221, 2xCyPET-3xFLAG-6xHIS/pDONRP2R-P3 (p3') and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|2xCyPet-HellFire 53|1 55|8530 56|10227 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|1 55|9436 56|10176 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|8698 56|9438 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Hellfire 53|1 55|8530 56|8697 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|8650 56|8667 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|3xFlag 53|1 55|8530 56|8595 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|8602 56|8622 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|10156 56|10185 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9418 56|9438 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|8680 56|8694 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PX domain p40phox 53|1 55|10285 56|10725 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|GAGA linker 53|1 55|10261 56|10284 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10756 56|11255 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11060 56|11144 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10756 56|11059 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11145 56|12741 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-3xFLAG-6xHIS/pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|1722 76|8516 77|10237 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xp40p221 72|0 73|0 74|1 75|504 76|10238 77|10741 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UBQ10prom/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10742 77|12751 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgccttgtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcacca 209|gggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatagccactttgtacaagaaagctgggtactcgagagaaccagcaccagcaccagcaccgagtgcctggggcacctgctctgagtcatagggcgactggtaaaagaagatccggacgtcctcatccatcagcacccagaccggcaggctgagcaggctcttcatgtaggcgttgagggcaggtatccgcatctcggcgatctcctgtttcacacccacgtagactttggctgggagtgtgggcagggtacaggccagggcactgctcttgctgtctggcccgaagcgctcctccagcttgctctgcaaagcatggaactggcggtagcggcggtagatgaggtacttggatcctccttttgtcttcacctcgatgacgaaaacaaagtggctggtgaagcctctcttctcctcgatgtcagcaatgttggccgagatggcaacatcatccggaagctgttcaaagtcactctcggcccgcagctgctgggccacagccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaa 209|agctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgagg 209|attttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P5C [UBQ10prom::2xCyPet-1xPH(FAPP1)/pK7m34GW] 27|0 222|3 33|12592 236|655480079 237|655480079 26|2756 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10575 56|10595 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8844 56|8864 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12585 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221, 1xFAPP1p3' and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PH domain FAPP1 53|1 55|8533 56|8835 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|1 55|9772 56|10512 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|9034 56|9774 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|8986 56|9003 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|8938 56|8958 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|10492 56|10521 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9754 56|9774 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9016 56|9030 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10597 56|11096 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10901 56|10985 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10597 56|10900 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|10986 56|12582 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xFAPP1p3' 72|0 73|0 74|1 75|336 76|8516 77|8851 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221 72|0 73|0 74|1 75|1731 76|8852 77|10582 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10583 77|12592 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatttagtccttgtatcagtcaaacatgctttggagctccccagagcgaccagccacctctgtctttcagctgcattcactgccttcatgtagaaatgctgctctccaggaatgattaattccattcttgtgttgtctgctgaatgaactttaatttcacaaactgccatctttatgcttcctttgctccctttgcaaacatcatcttgtgaatcatagtaggataagattccattatctaaaacaaaccaacgaggctgccagcctgtgagatagttggtccacttgtacaacaccccctccatctcgagagccactttgtacaagaaagctgggtagtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgccgt 209|gggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaacc 209|actttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P1C [UBQ10prom::2xCyPet-1xFYVE(HRS)/pK7m34GW] 27|0 222|3 33|12514 236|655480059 237|655480059 26|2755 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10497 56|10517 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8766 56|8786 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12507 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221, 1xFYVEp3' and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|FYVE 53|1 55|8533 56|8763 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|1 55|9694 56|10434 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|8956 56|9696 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|8908 56|8925 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|8860 56|8880 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|10414 56|10443 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9676 56|9696 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|8938 56|8952 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10519 56|11018 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10823 56|10907 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10519 56|10822 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|10908 56|12504 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xFYVEp3' 72|0 73|0 74|1 75|258 76|8516 77|8773 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221 72|0 73|0 74|1 75|1731 76|8774 77|10504 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10505 77|12514 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatgccttcttgttcagctgctcatagcagggctcacacacgcgcacctccttctcaatgccgaacttggggatggtggagtacttggaggagcacttgccacagaagatctgcccacatgctcggcagtgatgcttgcgggtcaccaccccaaactgtactctgcaccgatggcattcctcagcatccacccagtcaggggctctttcagcagcgaacatggcatcactttcagccactttgtacaagaaagctgggtagtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgc 209|tctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagattt 209|aacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P7C [UBQ10prom::2xCyPet-1xPH(OSBP)/pK7m34GW] 27|0 222|3 33|12595 236|655480037 237|655480037 26|2754 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10578 56|10598 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8847 56|8867 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12588 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221, 1xOSBPp3' and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PH domain OSBP human 53|1 55|8533 56|8838 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|1 55|9775 56|10515 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|9037 56|9777 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|8989 56|9006 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|8941 56|8961 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|10495 56|10524 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9757 56|9777 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9019 56|9033 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10600 56|11099 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10904 56|10988 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10600 56|10903 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|10989 56|12585 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xOSBPp3' 72|0 73|0 74|1 75|339 76|8516 77|8854 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221 72|0 73|0 74|1 75|1731 76|8855 77|10585 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10586 77|12595 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatgccagcatcttcacagctttggccttggccagttccagggccgtcacccagcgctgccgctcaacttctgaactagctttcagatggtaggtctgagcacccccattggaaatgatgaagttgcaggagtcctccacggtgatgttggctgtggcgaggttgatggtaccacggcaggtatgtctcatttctgcctttgatctgtagtagctcaggagcccgttgctcagcacgaaccatcgccgctggtagcctttgatataattggtccatttgaagagccagccctctcgagccgagcccgaagatctagccactttgtacaagaaagctgggtagtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgc 209|cgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgca 209|accactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P14C [UBQ10prom::2xCyPet-1xPH(PLC)/pK7m34GW] 27|0 222|3 33|12808 236|655480012 237|655480012 26|2753 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10791 56|10811 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9060 56|9080 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12801 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221, 1xPLCp3' and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|PH domain PLCd1 (Rat) 53|1 55|8533 56|9057 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|1 55|9988 56|10728 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|9250 56|9990 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|9202 56|9219 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|9154 56|9174 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|10708 56|10737 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9970 56|9990 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9232 56|9246 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10813 56|11312 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11117 56|11201 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10813 56|11116 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11202 56|12798 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xPLCp3' 72|0 73|0 74|1 75|552 76|8516 77|9067 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221 72|0 73|0 74|1 75|1731 76|9068 77|10798 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10799 77|12808 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactggatgttgagctccttcaggaagtccttcagctccttgaagttcatcttgttgtccttgtttttatcagcctttcgcaagcaggagtgaatccagtgctgcagcttctgccgctggtccatggagccggagtggtggatgatcttgcgcaggccctgcacccagtgctgagcgtcagctggtgatggggcaatgaggtctagggtgttgcgctggtccttgaagacaatggagaagcatcgatcctcggggatgtctcgggcaaacttctccaggccttctgtgcggtgtcccatccgtacctcctgaatgtcctcgatggagaacagctgcgactccggggacctcatgacctttcgagattcctgccagatggtcttgcagtcctcctgtagcttgtagaagcgttccctacgccacgagctggacttcaccttcagaagctggctgcccttcagaagggcctgaaggtccgggtcatcctggagcccgtgcagggtcaggaagtcccggcccgagtccatagccactttgtacaagaaagctgggtagtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggcc 209|agggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctcttt 209|gggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgct 209|tgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P15C [UBQ10prom::2xCyPet-1xTUBBY-C(TUBBY)/pK7m34GW] 27|0 222|3 33|13072 236|655479955 237|655479955 26|2752 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|11055 56|11075 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9324 56|9344 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|13065 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221, 1xTUBBYp3' and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|Tubby domain 53|1 55|8533 56|9321 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|1 55|10252 56|10992 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|9514 56|10254 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|9466 56|9483 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|9418 56|9438 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|10972 56|11001 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10234 56|10254 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9496 56|9510 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|11077 56|11576 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11381 56|11465 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|11077 56|11380 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11466 56|13062 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xTUBBYp3' 72|0 73|0 74|1 75|816 76|8516 77|9331 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221 72|0 73|0 74|1 75|1731 76|9332 77|11062 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|11063 77|13072 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactcgcaggccagcttgctgtcaaagctggacagagcaatggcaaaggcctgcagtgcacacagtgggtagttgtaatccatggtgaacacatcttctgctacccggccaaactgcatgacgatgtagtccgggtcattgccgtggatgatctggaagttcttcacagaagcctgtgtgacacggccgtggaagttaagtacataggactgtgtgtcatcattccagactggcgtcttgttctgcagctcaatgatgctctccgtgttcttgttctgccagcgtgctaacagggtctcatgttcattgcgggggcggatacagactctctcatgaaccatgttcatgcctgggacgatcacactcatcttccgaggtcccttgaagcctaggacatttgtctcatagcacaccgctgccagctcctggcgcaaggttccgctttccagcgtggaagaggatgccttctgagggttgacgccattgtcataaacggtgaacttggtgcccatcaggttggaccgcaatttcccgatatagctatcgcctccccgagacaagtctgttgggtccacagagatgaggtaattggaagttttactcttctttctcttcctgcccgccaggaggaacaccttcttgccatcctcacggtctaggtgcagaaagtaggtggggtacatgccgcggtccatccccttcttgtcccgagtgatgcggcatttgatggtgatcccttgtggggctggcctcagtgcaaactcctctagatcctggacctcaatatccacagccactttgtacaagaaagctgggtagtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgt 209|agttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaa 209|caaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccac 209|cacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P23C [UBQ10prom::2xCyPet-2xPH(OSBP)/pK7m34GW] 27|0 222|3 33|12934 236|655479927 237|655479927 26|2751 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10917 56|10937 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9186 56|9206 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12927 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221, 2xOSBPp3' and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|KGAGAGAGS linker 53|1 55|8845 56|8871 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain OSBP human 53|1 55|8872 56|9177 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain OSBP human 53|1 55|8533 56|8838 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|1 55|10114 56|10854 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|9376 56|10116 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|9328 56|9345 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|9280 56|9300 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|10834 56|10863 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10096 56|10116 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9358 56|9372 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10939 56|11438 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11243 56|11327 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10939 56|11242 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11328 56|12924 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xOSBPp3' 72|0 73|0 74|1 75|678 76|8516 77|9193 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221 72|0 73|0 74|1 75|1731 76|9194 77|10924 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10925 77|12934 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatgccagcatcttcacagctttggccttggccagttccagggccgtcacccagcgctgccgctcaacttctgaactagctttcagatggtaggtctgagcacccccattggaaatgatgaagttgcaggagtcctccacggtgatgttggctgtggcgaggttgatggtaccacggcaggtatgtctcatttctgcctttgatctgtagtagctcaggagcccgttgctcagcacgaaccatcgccgctggtagcctttgatataattggtccatttgaagagccagccctctcgagccgagcccgaagatctagaaccagcaccagcaccagcaccctttgccagcatcttcacagctttggccttggccagttccagggccgtcacccagcgctgccgctcaacttctgaactagctttcagatggtaggtctgagcacccccattggaaatgatgaagttgcaggagtcctccacggtgatgttggctgtggcgaggttgatggtaccacggcaggtatgtctcatttctgcctttgatctgtagtagctcaggagcccgttgctcagcacgaaccatcgccgctggtagcctttgatataattggtccatttgaagagccagccctctcgagccgagcccgaagatctagccactttgtacaagaaagctgggtagtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaaga 209|agatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaac 209|aaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcg 209|tgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P25C [UBQ10prom::2xCyPet-2xTUBBY-C(TUBBY)/pK7m34GW] 27|0 222|3 33|13894 236|655479892 237|655479892 26|2750 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|11877 56|11897 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|10146 56|10166 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|13887 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221, 2xTubbp3' and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PolyA liner 53|1 55|9328 56|9348 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TubbyC 53|1 55|9349 56|10137 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TubbyC 53|1 55|8533 56|9321 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|1 55|11074 56|11814 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|10336 56|11076 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|10288 56|10305 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|10240 56|10260 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|11794 56|11823 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|11056 56|11076 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10318 56|10332 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|11899 56|12398 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|12203 56|12287 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|11899 56|12202 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|12288 56|13884 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xTubbp3' 72|0 73|0 74|1 75|1638 76|8516 77|10153 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221 72|0 73|0 74|1 75|1731 76|10154 77|11884 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|11885 77|13894 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactcgcaggccagcttgctgtcaaagctggacagagcaatggcaaaggcctgcagtgcacacagtgggtagttgtaatccatggtgaacacatcttctgctacccggccaaactgcatgacgatgtagtccgggtcattgccgtggatgatctggaagttcttcacagaagcctgtgtgacacggccgtggaagttaagtacataggactgtgtgtcatcattccagactggcgtcttgttctgcagctcaatgatgctctccgtgttcttgttctgccagcgtgctaacagggtctcatgttcattgcgggggcggatacagactctctcatgaaccatgttcatgcctgggacgatcacactcatcttccgaggtcccttgaagcctaggacatttgtctcatagcacaccgctgccagctcctggcgcaaggttccgctttccagcgtggaagaggatgccttctgagggttgacgccattgtcataaacggtgaacttggtgcccatcaggttggaccgcaatttcccgatatagctatcgcctccccgagacaagtctgttgggtccacagagatgaggtaattggaagttttactcttctttctcttcctgcccgccaggaggaacaccttcttgccatcctcacggtctaggtgcagaaagtaggtggggtacatgccgcggtccatccccttcttgtcccgagtgatgcggcatttgatggtgatcccttgtggggctggcctcagtgcaaactcctctagatcctggacctcaatatccacagatctagcggcagcagcagcatctagctcgcaggccagcttgctgtcaaagctggacagagcaatggcaaaggcctgcagtgcacacagtgggtagttgtaatccatggtgaacacatcttctgctacccggccaaactgcatgacgatgtagtccgggtcattgccgtggatgatctggaagttcttcacagaagcctgtgtgacacggccgtggaagttaagtacataggactgtgtgtcatcattccagactggcgtcttgttctgcagctcaatgatgctctccgtgttcttgttctgccagcgtgctaacagggtctcatgttcattgcgggggcggatacagactctctcatgaaccatgttcatgcctgggacgatcacactcatcttccgaggtcccttgaagcctaggacatttgtctcatagcacaccgctgccagctcctggcgcaaggttccgctttccagcgtgg 209|aagaggatgccttctgagggttgacgccattgtcataaacggtgaacttggtgcccatcaggttggaccgcaatttcccgatatagctatcgcctccccgagacaagtctgttgggtccacagagatgaggtaattggaagttttactcttctttctcttcctgcccgccaggaggaacaccttcttgccatcctcacggtctaggtgcagaaagtaggtggggtacatgccgcggtccatccccttcttgtcccgagtgatgcggcatttgatggtgatcccttgtggggctggcctcagtgcaaactcctctagatcctggacctcaatatccacctcgagagccactttgtacaagaaagctgggtagtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtgg 209|ttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttata 209|tttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P5R [UBQ10prom::2xCHERRY-1xPH(FAPP1)/pH7m34GW] 27|0 222|3 33|12823 236|655479088 237|655479088 26|2749 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10806 56|10826 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9231 56|9251 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12816 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCHERRY-1xmyc/pDONR221, 1xFAPP1p3' and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PH domain FAPP1 53|1 55|8920 56|9222 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|9322 56|10026 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|10060 56|10764 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10027 56|10050 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|10765 56|10788 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY 53|1 55|9322 56|10794 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|9259 56|9288 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|9298 56|9312 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-1xMyc 53|1 55|9259 56|10794 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10828 56|11327 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11132 56|11216 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10828 56|11131 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11217 56|12813 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xFAPP1p3' 72|0 73|0 74|1 75|336 76|8903 77|9238 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCHERRY-1xmyc/pDONR221 72|0 73|0 74|1 75|1575 76|9239 77|10813 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10814 77|12823 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatttagtccttgtatcagtcaaacatgctttggagctccccagagcgaccagccacctctgtctttcagctgcattcactgccttcatgtagaaatgctgctctccaggaatgattaattccattcttgtgttgtctgctgaatgaactttaatttcacaaactgccatctttatgcttcctttgctccctttgcaaacatcatcttgtgaatcatagtaggataagattccattatctaaaacaaaccaacgaggctgccagcctgtgagatagttggtccacttgtacaacaccccctccatctcgagagccactttgtacaagaaagctgggtaattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgt 209|aggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaa 209|gaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggtta 209|catgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P7R [UBQ10prom::2xCHERRY-1xPH(OSBP)/pH7m34GW]' 27|0 222|3 33|12826 236|655479066 237|655479066 26|2748 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10809 56|10829 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9234 56|9254 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12819 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCHERRY-1xmyc/pDONR221, 1xOSBPp3' and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PH domain OSBP human 53|1 55|8920 56|9225 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|9325 56|10029 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|10063 56|10767 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10030 56|10053 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|10768 56|10791 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY 53|1 55|9325 56|10797 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|9262 56|9291 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|9301 56|9315 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-1xMyc 53|1 55|9262 56|10797 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10831 56|11330 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11135 56|11219 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10831 56|11134 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11220 56|12816 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xOSBPp3' 72|0 73|0 74|1 75|339 76|8903 77|9241 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCHERRY-1xmyc/pDONR221 72|0 73|0 74|1 75|1575 76|9242 77|10816 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10817 77|12826 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatgccagcatcttcacagctttggccttggccagttccagggccgtcacccagcgctgccgctcaacttctgaactagctttcagatggtaggtctgagcacccccattggaaatgatgaagttgcaggagtcctccacggtgatgttggctgtggcgaggttgatggtaccacggcaggtatgtctcatttctgcctttgatctgtagtagctcaggagcccgttgctcagcacgaaccatcgccgctggtagcctttgatataattggtccatttgaagagccagccctctcgagccgagcccgaagatctagccactttgtacaagaaagctgggtaattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttca 209|cgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgt 209|gaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttgg 209|ttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P14R [UBQ10prom::2xCHERRY-1xPH(PLC)/pH7m34GW] 27|0 222|3 33|13039 236|655479041 237|655479041 26|2747 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|11022 56|11042 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9447 56|9467 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|13032 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCHERRY-1xmyc/pDONR221, 1xPLCp3' and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|PH domain PLCd1 (Rat) 53|1 55|8920 56|9444 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|9538 56|10242 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|10276 56|10980 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10243 56|10266 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|10981 56|11004 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY 53|1 55|9538 56|11010 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|9475 56|9504 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|9514 56|9528 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-1xMyc 53|1 55|9475 56|11010 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|11044 56|11543 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11348 56|11432 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|11044 56|11347 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11433 56|13029 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xPLCp3' 72|0 73|0 74|1 75|552 76|8903 77|9454 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCHERRY-1xmyc/pDONR221 72|0 73|0 74|1 75|1575 76|9455 77|11029 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|11030 77|13039 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactggatgttgagctccttcaggaagtccttcagctccttgaagttcatcttgttgtccttgtttttatcagcctttcgcaagcaggagtgaatccagtgctgcagcttctgccgctggtccatggagccggagtggtggatgatcttgcgcaggccctgcacccagtgctgagcgtcagctggtgatggggcaatgaggtctagggtgttgcgctggtccttgaagacaatggagaagcatcgatcctcggggatgtctcgggcaaacttctccaggccttctgtgcggtgtcccatccgtacctcctgaatgtcctcgatggagaacagctgcgactccggggacctcatgacctttcgagattcctgccagatggtcttgcagtcctcctgtagcttgtagaagcgttccctacgccacgagctggacttcaccttcagaagctggctgcccttcagaagggcctgaaggtccgggtcatcctggagcccgtgcagggtcaggaagtcccggcccgagtccatagccactttgtacaagaaagctgggtaattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcct 209|cccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcga 209|gaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatgggg 209|catggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P3R [UBQ10prom::1xPX(p40)-2xCHERRY/pH7m34GW] 27|0 222|3 33|13150 236|655478830 237|655478830 26|2746 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|11133 56|11153 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|10629 56|10649 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|13143 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 1xp40p221, 2xmCHERRY-4xMyc/pDONRP2R-P3 (p3') and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|mCHERRY 53|1 55|9839 56|10575 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|9101 56|9837 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|4xMyc 53|1 55|8920 56|9078 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10546 56|10560 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9808 56|9822 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9079 56|9093 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|9040 56|9069 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|9001 56|9030 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|8962 56|8991 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|8923 56|8952 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-4xMyc 53|1 55|8920 56|10626 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PX domain p40phox 53|1 55|10684 56|11124 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|GAGA linker 53|1 55|10660 56|10683 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|11155 56|11654 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11459 56|11543 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|11155 56|11458 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11544 56|13140 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xmCHERRY-4xMyc/pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|1734 76|8903 77|10636 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xp40p221 72|0 73|0 74|1 75|504 76|10637 77|11140 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|11141 77|13150 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgctcaattcaagtcctcttcagaaatgagcttttgctccatttcattcaagtcctcttcagaaatgagcttttgctccatttcattcaagtcctcttcagaaatgagcttttgctccatttcattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgccct 209|tgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatagccactttgtacaagaaagctgggtactcgagagaaccagcaccagcaccagcaccgagtgcctggggcacctgctctgagtcatagggcgactggtaaaagaagatccggacgtcctcatccatcagcacccagaccggcaggctgagcaggctcttcatgtaggcgttgagggcaggtatccgcatctcggcgatctcctgtttcacacccacgtagactttggctgggagtgtgggcagggtacaggccagggcactgctcttgctgtctggcccgaagcgctcctccagcttgctctgcaaagcatggaactggcggtagcggcggtagatgaggtacttggatcctccttttgtcttcacctcgatgacgaaaacaaagtggctggtgaagcctctcttctcctcgatgtcagcaatgttggccgagatggcaacatcatccggaagctgttcaaagtcactctcggcccgcagctgctgggccacagccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcaca 209|aactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattag 209|aaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P13Y [UBQ10prom::1xENTH(ent5p)-CITRINE/pB7m34GW] 27|0 222|3 33|11556 236|655478321 237|655478321 26|2745 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9539 56|9559 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8999 56|9019 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11549 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 1xEnt5p221, mCITRINE+STOP/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|mCITRINE 53|1 55|8280 56|8996 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|ENTH domain Ent5p 53|1 55|9024 56|9530 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9561 56|10060 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9865 56|9949 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9561 56|9864 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9950 56|11546 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINE+STOP/pDONRP2R-P3 72|0 73|0 74|1 75|744 76|8263 77|9006 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xEnt5p221 72|0 73|0 74|1 75|540 76|9007 77|9546 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9547 77|11556 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcttacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtgg 209|ttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatagccactttgtacaagaaagctgggtaaagttctccctttttcctaatacctctctcatgaatctccattttaccatcattggagaattttaccttgaattgggcaatctccctggttaaaatatgcttatgagtgagcaaacatgatctaatctgattcagttcgtcacctgtgtcgacgttcaataataggaattcaatgactactagacattttaagaccaccctccattcggacccataattaacgtagtcctttcttgccttttcgtataggttcttgggcctggtagcgatatggtcgactaatcttttcaacgtgtattctgtcatcaagtagagcggcacttggtatctgttccttaaaaccttggcgaggtgcttcggtgtggggccccaagcatcggtatttgtagcacgacggatatctatctgataaggctcatattggactatcacgttttgggcaaaccttgcggcatttctaatatcatggatacctaaattttgaatcttttttgataatgagtccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatac 209|gaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaaga 209|gctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P7Y [UBQ10prom::CITRINE-1xPH(OSBP)/pB7m34GW] 27|0 222|3 33|11361 236|655478095 237|655478095 26|2744 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9344 56|9364 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8594 56|8614 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11354 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 1xOSBPp3' and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PH domain OSBP human 53|1 55|8280 56|8585 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8619 56|9335 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9366 56|9865 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9670 56|9754 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9366 56|9669 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9755 56|11351 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xOSBPp3' 72|0 73|0 74|1 75|339 76|8263 77|8601 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8602 77|9351 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9352 77|11361 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatgccagcatcttcacagctttggccttggccagttccagggccgtcacccagcgctgccgctcaacttctgaactagctttcagatggtaggtctgagcacccccattggaaatgatgaag 209|ttgcaggagtcctccacggtgatgttggctgtggcgaggttgatggtaccacggcaggtatgtctcatttctgcctttgatctgtagtagctcaggagcccgttgctcagcacgaaccatcgccgctggtagcctttgatataattggtccatttgaagagccagccctctcgagccgagcccgaagatctagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactag 209|gtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagc 209|aagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P4Y [UBQ10prom::CITRINE-1xPH(OSH2)/pB7m34GW] 27|0 222|3 33|11553 236|655478069 237|655478069 26|2743 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9536 56|9556 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8786 56|8806 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11546 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 1xOSH2p3' and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PH domain OSH2p 53|1 55|8280 56|8783 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8811 56|9527 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9558 56|10057 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9862 56|9946 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9558 56|9861 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9947 56|11543 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xOSH2p3' 72|0 73|0 74|1 75|531 76|8263 77|8793 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8794 77|9543 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9544 77|11553 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatagagatccttgtgttttggaatgcgatatcagggctggagatttgttcttcaaggccaatgatggtggtactgccttttttttgtttaatatctccttgtctttggcgaatctaatagcg 209|ctttgtatagcattgacccacctagtcgtttcaatgggatgattgccctttaagcgccaccttgttgtgcctgtgatgccacctagcaactcgaaattcagtttttctgaagaatcgatgtgtaagcggcacgtcgaaacttttaaagtaccacgaggcctatcaacatgcgattggtctttgtaatatgacagatttccgtcgccacttaaaatgaaccaacgtaatttgtagccatgggcaaaatttgtccatttctttaaaaaccccttatacgtaggaggcttacttgatgcgacatcttcgtgaacaatagcttgttctctcagattcttttcaagtagattctttaacttaatttccggtgtaacgttgttcgacggagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaa 209|attgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagct 209|gtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P19Y [UBQ10prom::CITRINE-2xPH(AtPH1)/pB7m34GW] 27|0 222|3 33|11724 236|655478042 237|655478042 26|2742 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9707 56|9727 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8957 56|8977 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11717 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 2xAtPH1p3' and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|KGAGAGAGS linker 53|1 55|8604 56|8630 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|PH domain AtPH1 53|1 55|8603 56|8953 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|PH domain AtPH1 53|1 55|8280 56|8597 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8982 56|9698 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9729 56|10228 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10033 56|10117 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9729 56|10032 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|10118 56|11714 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xAtPH1p3' 72|0 73|0 74|1 75|702 76|8263 77|8964 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8965 77|9714 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9715 77|11724 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctattgcacaatcgatcttccaatcgaattaatccactcttctttctccttctcattatcagcaatgaagaacatggtatagctaccactagatagctcaaaagcaaaaggcttattcacaaca 209|tcctcagctcctttcacggtgagacaatcaccaacggagatcacaccacgcggcgtagatccacgaattccagcagcggcttgatctttgaaccagagaagcttccctcgtttgagaacgaaccaacgacgacgccaggttttgatgtaatcgccttgctttgtgagccaaccagaacgctcagggtttgaccagaactcgagagaaccagcaccagcaccagcacccttttgcacaatcgatcttccaatcgaattaatccactcttctttctccttctcattatcagcaatgaagaacatggtatagctaccactagatagctcaaaagcaaaaggcttattcacaacatcctcagctcctttcacggtgagacaatcaccaacggagatcacaccacgcggcgtagatccacgaattccagcagcggcttgatctttgaaccagagaagcttccctcgtttgagaacgaaccaacgacgacgccaggttttgatgtaatcgccttgctttgtgagccaaccagaacgctcagggtttgaccagaactcgagagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctaga 209|tagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaagga 209|aagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P21Y [UBQ10prom::CITRINE-2xPH(FAPP1)/pB7m34GW] 27|0 222|3 33|11694 236|655477970 237|655477970 26|2741 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9677 56|9697 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8927 56|8947 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11687 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 2xFAPP1p3' and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|15 52|KGAGAGAGS linker 53|1 55|8589 56|8615 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain FAPP1 53|1 55|8616 56|8918 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain FAPP1 53|1 55|8280 56|8582 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8952 56|9668 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9699 56|10198 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10003 56|10087 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9699 56|10002 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|10088 56|11684 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xFAPP1p3' 72|0 73|0 74|1 75|672 76|8263 77|8934 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8935 77|9684 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9685 77|11694 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatttagtccttgtatcagtcaaacatgctttggagctccccagagcgaccagccacctctgtctttcagctgcattcactgccttcatgtagaaatgctgctctccaggaatgattaattcc 209|attcttgtgttgtctgctgaatgaactttaatttcacaaactgccatctttatgcttcctttgctccctttgcaaacatcatcttgtgaatcatagtaggataagattccattatctaaaacaaaccaacgaggctgccagcctgtgagatagttggtccacttgtacaacaccccctccatctcgagagaaccagcaccagcaccagcaccctttttagtccttgtatcagtcaaacatgctttggagctccccagagcgaccagccacctctgtctttcagctgcattcactgccttcatgtagaaatgctgctctccaggaatgattaattccattcttgtgttgtctgctgaatgaactttaatttcacaaactgccatctttatgcttcctttgctccctttgcaaacatcatcttgtgaatcatagtaggataagattccattatctaaaacaaaccaacgaggctgccagcctgtgagatagttggtccacttgtacaacaccccctccatctcgagagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattc 209|gacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacaga 209|cgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P18Y [UBQ10prom::CITRINE-2xFYVE(HRS)/pB7m34GW] 27|0 222|3 33|11526 236|655477935 237|655477935 26|2740 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9509 56|9529 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8759 56|8779 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11519 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 2xFYVEp3' and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|6 52|FYVE 53|1 55|8526 56|8756 57|1 281|1 282|1 283|1 284|1 285|3363..3593 50 45 51|6 52|FYVE 53|1 55|8280 56|8510 57|1 281|1 282|1 283|1 284|1 285|3609..3839 50 45 51|4 52|mCITRINE 53|1 55|8784 56|9500 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9531 56|10030 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9835 56|9919 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9531 56|9834 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9920 56|11516 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xFYVEp3' 72|0 73|0 74|1 75|504 76|8263 77|8766 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8767 77|9516 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9517 77|11526 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgctagtgccttcttgttcagctgctcatagcagggctcacacacgcgcacctccttctcaatgccgaacttggggatggtggagtacttggaggagcacttgccacagaagatctgcccacatgct 209|cggcagtgatgcttgcgggtcaccaccccaaactgtactctgcaccgatggcattcctcagcatccacccagtcaggggctctttcagcagcgaacatggcatcactttcggatccttgtccttgtgccttcttgttcagctgctcatagcagggctcacacacgcgcacctccttctcaatgccgaacttggggatggtggagtacttggaggagcacttgccacagaagatctgcccacatgctcggcagtgatgcttgcgggtcaccaccccaaactgtactctgcaccgatggcattcctcagcatccacccagtcaggggctctttcagcagcgaacatggcatcactttcagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcga 209|agattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaa 209|gtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P22Y [UBQ10prom::CITRINE-2xGRAM(Atg26)/pB7m34GW] 27|0 222|3 33|11490 236|655477901 237|655477901 26|2739 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9473 56|9493 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8723 56|8743 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11483 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 2xGRAMp3' and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|KGAGAGAGS linker 53|1 55|8487 56|8513 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|GRAM domain 53|1 55|8514 56|8714 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|GRAM domain 53|1 55|8280 56|8480 57|1 281|1 282|1 283|1 284|1 50 45 51|62 52|Y57P 53|1 54|Y57P when the conserved Tyr57 is mutated into a Proline, the GRAM domain lose its ability to bind PI4P 55|8310 56|8312 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8748 56|9464 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9495 56|9994 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9799 56|9883 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9495 56|9798 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9884 56|11480 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xGRAMp3' 72|0 73|0 74|1 75|468 76|8263 77|8730 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8731 77|9480 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9481 77|11490 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctacttcaaattataaacattctcaatatcggagtatggaagtatcatagttgtgtttgtgccgggaaataaagatctgtagcaaatgcaagttgttcctaggtatactttgccgtatacaggt 209|atatttttttgaaaatggcagaagtaactagccagtagctcctctgaatcaggaagggaaaaatgctttcgaaatctggactcgagagaaccagcaccagcaccagcacccttcttcaaattataaacattctcaatatcggagtatggaagtatcatagttgtgtttgtgccgggaaataaagatctgtagcaaatgcaagttgttcctaggtatactttgccgtatacaggtatatttttttgaaaatggcagaagtaactagccagtagctcctctgaatcaggaagggaaaaatgctttcgaaatctggactcgagagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttacct 209|tgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtca 209|ttatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P20Y [UBQ10prom::CITRINE-2xPH(OSH2)/pB7m34GW] 27|0 222|3 33|12072 236|655477858 237|655477858 26|2738 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10055 56|10075 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9305 56|9325 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12065 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 2xOSH2p3' and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|PH domain OSH2p 53|1 55|8781 56|9302 57|1 281|1 282|1 283|1 284|1 50 45 51|45 52|VNSKL linker 53|1 55|8781 56|8801 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|PH domain OSH2 53|1 55|8280 56|8780 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|9330 56|10046 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10077 56|10576 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10381 56|10465 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10077 56|10380 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|10466 56|12062 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xOSH2p3' 72|0 73|0 74|1 75|1050 76|8263 77|9312 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|9313 77|10062 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10063 77|12072 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctaagatccttgtgttttggaatgcgatatcagggctggagatttgttcttcaaggccaatgatggtggtactgccttttttttgtttaatatctccttgtctttggcgaatctaatagcgctt 209|tgtatagcattgacccacctagtcgtttcaatgggatgattgccctttaagcgccaccttgttgtgcctgtgatgccacctagcaactcgaaattcagtttttctgaagaatcgatgtgtaagcggcacgtcgaaacttttaaagtaccacgaggcctatcaacatgcgattggtctttgtaatatgacagatttccgtcgccacttaaaatgaaccaacgtaatttgtagccatgggcaaaatttgtccatttctttaaaaaccccttatacgtaggaggcttacttgatgcgacatcttcgtgaacaatagcttgttctctcagattcttttcaagtagattctttaacttaatttccggtgtaacgttgttcgacggaagcttcgaattcaccggtagagatccttgtgttttggaatgcgatatcagggctggagatttgttcttcaaggccaatgatggtggtactgccttttttttgtttaatatctccttgtctttggcgaatctaatagcgctttgtatagcattgacccacctagtcgtttcaatgggatgattgccctttaagcgccaccttgttgtgcctgtgatgccacctagcaactcgaaattcagtttttctgaagaatcgatgtgtaagcggcacgtcgaaacttttaaagtaccacgaggcctatcaacatgcgattggtctttgtaatatgacagatttccgtcgccacttaaaatgaaccaacgtaatttgtagccatgggcaaaatttgtccatttctttaaaaaccccttatacgtaggaggcttacttgatgcgacatcttcgtgaacaatagcttgttctctcagattcttttcaagtagattctttaacttaatttccggtgtaacgttgttcgacggagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtc 209|gtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatat 209|ttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P24Y [UBQ10prom::CITRINE-2xPH(PLC)/pB7m34GW] 27|0 222|3 33|12117 236|655477815 237|655477815 26|2737 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10100 56|10120 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9350 56|9370 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12110 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 2xPLCp3' and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|2xPH domain PLCd1 53|1 55|8280 56|9347 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain PLCd1 Rat 53|1 55|8823 56|9347 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain PLCd1 Rat 53|1 55|8280 56|8791 57|1 281|1 282|1 283|1 284|1 50 45 51|84 52|GAGARS linker 53|1 55|8792 56|8822 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|9375 56|10091 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10122 56|10621 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10426 56|10510 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10122 56|10425 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|10511 56|12107 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xPLCp3' 72|0 73|0 74|1 75|1095 76|8263 77|9357 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|9358 77|10107 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10108 77|12117 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactggatgttgagctccttcaggaagtccttcagctccttgaagttcatcttgttgtccttgtttttatcagcctttcgcaagcaggagtgaatccagtgctgcagcttctgccgctggtcc 209|atggagccggagtggtggatgatcttgcgcaggccctgcacccagtgctgagcgtcagctggtgatggggcaatgaggtctagggtgttgcgctggtccttgaagacaatggagaagcatcgatcctcggggatgtctcgggcaaacttctccaggccttctgtgcggtgtcccatccgtacctcctgaatgtcctcgatggagaacagctgcgactccggggacctcatgacctttcgagattcctgccagatggtcttgcagtcctcctgtagcttgtagaagcgttccctacgccacgagctggacttcaccttcagaagctggctgcccttcagaagggcctgaaggtccgggtcatcctggagcccgtgcagggtcaggaagtccctacccgagtccatggatctagctcctgcaccctggatgttgagctccttcaggaagtccttcagctccttgaagttcatcttgttgtccttgtttttatcagcctttcgcaagcaggagtgaatccagtgctgcagcttctgccgctggtccatggagccggagtggtggatgatcttgcgcaggccctgcacccagtgctgagcgtcagctggtgatggggcaatgaggtctagggtgttgcgctggtccttgaagacaatggagaagcatcgatcctcggggatgtctcgggcaaacttctccaggccttctgtgcggtgtcccatccgtacctcctgaatgtcctcgatggagaacagctgcgactccggggacctcatgacctttcgagattcctgccagatggtcttgcagtcctcctgtagcttgtagaagcgttccctacgccacgagctggacttcaccttcagaagctggctgcccttcagaagggcctgaaggtccgggtcatcctggagcccgtgcagggtcaggaagtcccggcccgagtccatagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggt 209|gcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgtta 209|tttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P23R [UBQ10prom::2xCHERRY-1xPH(OSBP)/pH7m34GW] 27|0 222|3 33|13165 236|628940119 237|628940119 26|2545 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|11148 56|11168 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9573 56|9593 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|13158 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCHERRY-1xmyc/pDONR221, 2xPH domain OSBP human/pDONRP2R-P3 and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|KGAGAGAGS linker 53|1 55|9232 56|9258 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain OSBP human 53|1 55|9259 56|9564 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain OSBP human 53|1 55|8920 56|9225 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|9664 56|10368 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|10402 56|11106 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10369 56|10392 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|11107 56|11130 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY 53|1 55|9664 56|11136 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|9601 56|9630 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|9640 56|9654 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-1xMyc 53|1 55|9601 56|11136 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|11170 56|11669 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11474 56|11558 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|11170 56|11473 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11559 56|13155 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xPH domain OSBP human/pDONRP2R-P3 72|0 73|0 74|1 75|678 76|8903 77|9580 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCHERRY-1xmyc/pDONR221 72|0 73|0 74|1 75|1575 76|9581 77|11155 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|11156 77|13165 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatgccagcatcttcacagctttggccttggccagttccagggccgtcacccagcgctgccgctcaacttctgaactagctttcagatggtaggtctgagcacccccattggaaatgatgaagttgcaggagtcctccacggtgatgttggctgtggcgaggttgatggtaccacggcaggtatgtctcatttctgcctttgatctgtagtagctcaggagcccgttgctcagcacgaaccatcgccgctggtagcctttgatataattggtccatttgaagagccagccctctcgagccgagcccgaagatctagaaccagcaccagcaccagcaccctttgccagcatcttcacagctttggccttggccagttccagggccgtcacccagcgctgccgctcaacttctgaactagctttcagatggtaggtctgagcacccccattggaaatgatgaagttgcaggagtcctccacggtgatgttggctgtggcgaggttgatggtaccacggcaggtatgtctcatttctgcctttgatctgtagtagctcaggagcccgttgctcagcacgaaccatcgccgctggtagcctttgatataattggtccatttgaagagccagccctctcgagccgagcccgaagatctagccactttgtacaagaaagctgggtaattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgg 209|gcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgac 209|aaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgc 209|agatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P25R [UBQ10prom::2xCHERRY-2xTUBBY-C(TUBBY)/pH7m34GW] 27|0 222|3 33|14125 236|628940066 237|628940066 26|2544 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|12108 56|12128 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|10533 56|10553 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|14118 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCHERRY-1xmyc/pDONR221, 2xTubbyC/pDONRP2R-P3 (p3') and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PolyA liner 53|1 55|9715 56|9735 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TubbyC 53|1 55|9736 56|10524 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TubbyC 53|1 55|8920 56|9708 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|10624 56|11328 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|11362 56|12066 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|11329 56|11352 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|12067 56|12090 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY 53|1 55|10624 56|12096 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|10561 56|10590 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|10600 56|10614 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-1xMyc 53|1 55|10561 56|12096 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|12130 56|12629 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|12434 56|12518 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|12130 56|12433 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|12519 56|14115 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xTubbyC/pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|1638 76|8903 77|10540 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCHERRY-1xmyc/pDONR221 72|0 73|0 74|1 75|1575 76|10541 77|12115 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|12116 77|14125 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactcgcaggccagcttgctgtcaaagctggacagagcaatggcaaaggcctgcagtgcacacagtgggtagttgtaatccatggtgaacacatcttctgctacccggccaaactgcatgacgatgtagtccgggtcattgccgtggatgatctggaagttcttcacagaagcctgtgtgacacggccgtggaagttaagtacataggactgtgtgtcatcattccagactggcgtcttgttctgcagctcaatgatgctctccgtgttcttgttctgccagcgtgctaacagggtctcatgttcattgcgggggcggatacagactctctcatgaaccatgttcatgcctgggacgatcacactcatcttccgaggtcccttgaagcctaggacatttgtctcatagcacaccgctgccagctcctggcgcaaggttccgctttccagcgtggaagaggatgccttctgagggttgacgccattgtcataaacggtgaacttggtgcccatcaggttggaccgcaatttcccgatatagctatcgcctccccgagacaagtctgttgggtccacagagatgaggtaattggaagttttactcttctttctcttcctgcccgccaggaggaacaccttcttgccatcctcacggtctaggtgcagaaagtaggtggggtacatgccgcggtccatccccttcttgtcccgagtgatgcggcatttgatggtgatcccttgtggggctggcctcagtgcaaactcctctagatcctggacctcaatatccacagatctagcggcagcagcagcatctagctcgcaggccagcttgctgtcaaagctggacagagcaatggcaaaggcctgcagtgcacacagtg 209|ggtagttgtaatccatggtgaacacatcttctgctacccggccaaactgcatgacgatgtagtccgggtcattgccgtggatgatctggaagttcttcacagaagcctgtgtgacacggccgtggaagttaagtacataggactgtgtgtcatcattccagactggcgtcttgttctgcagctcaatgatgctctccgtgttcttgttctgccagcgtgctaacagggtctcatgttcattgcgggggcggatacagactctctcatgaaccatgttcatgcctgggacgatcacactcatcttccgaggtcccttgaagcctaggacatttgtctcatagcacaccgctgccagctcctggcgcaaggttccgctttccagcgtggaagaggatgccttctgagggttgacgccattgtcataaacggtgaacttggtgcccatcaggttggaccgcaatttcccgatatagctatcgcctccccgagacaagtctgttgggtccacagagatgaggtaattggaagttttactcttctttctcttcctgcccgccaggaggaacaccttcttgccatcctcacggtctaggtgcagaaagtaggtggggtacatgccgcggtccatccccttcttgtcccgagtgatgcggcatttgatggtgatcccttgtggggctggcctcagtgcaaactcctctagatcctggacctcaatatccacctcgagagccactttgtacaagaaagctgggtaattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccc 209|tcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaa 209|taataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccga 209|gaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P25Y [UBQ10prom::CITRINE-2xTUBBY-C(TUBBY)/pB7m34GW 27|0 222|3 33|12660 236|628939979 237|628939979 26|2543 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10643 56|10663 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9893 56|9913 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12653 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 2xTubbyC/pDONRP2R-P3 (p3') and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PolyA liner 53|1 55|9075 56|9095 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TubbyC 53|1 55|9096 56|9884 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TubbyC 53|1 55|8280 56|9068 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|9918 56|10634 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10665 56|11164 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10969 56|11053 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10665 56|10968 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11054 56|12650 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xTubbyC/pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|1638 76|8263 77|9900 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|9901 77|10650 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10651 77|12660 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactcgcaggccagcttgctgtcaaagctggacagagcaatggcaaaggcctgcagtgcacacagtgggtagttgtaatccatggtgaacacatcttctgctacccggccaaactgcatgacg 209|atgtagtccgggtcattgccgtggatgatctggaagttcttcacagaagcctgtgtgacacggccgtggaagttaagtacataggactgtgtgtcatcattccagactggcgtcttgttctgcagctcaatgatgctctccgtgttcttgttctgccagcgtgctaacagggtctcatgttcattgcgggggcggatacagactctctcatgaaccatgttcatgcctgggacgatcacactcatcttccgaggtcccttgaagcctaggacatttgtctcatagcacaccgctgccagctcctggcgcaaggttccgctttccagcgtggaagaggatgccttctgagggttgacgccattgtcataaacggtgaacttggtgcccatcaggttggaccgcaatttcccgatatagctatcgcctccccgagacaagtctgttgggtccacagagatgaggtaattggaagttttactcttctttctcttcctgcccgccaggaggaacaccttcttgccatcctcacggtctaggtgcagaaagtaggtggggtacatgccgcggtccatccccttcttgtcccgagtgatgcggcatttgatggtgatcccttgtggggctggcctcagtgcaaactcctctagatcctggacctcaatatccacagatctagcggcagcagcagcatctagctcgcaggccagcttgctgtcaaagctggacagagcaatggcaaaggcctgcagtgcacacagtgggtagttgtaatccatggtgaacacatcttctgctacccggccaaactgcatgacgatgtagtccgggtcattgccgtggatgatctggaagttcttcacagaagcctgtgtgacacggccgtggaagttaagtacataggactgtgtgtcatcattccagactggcgtcttgttctgcagctcaatgatgctctccgtgttcttgttctgccagcgtgctaacagggtctcatgttcattgcgggggcggatacagactctctcatgaaccatgttcatgcctgggacgatcacactcatcttccgaggtcccttgaagcctaggacatttgtctcatagcacaccgctgccagctcctggcgcaaggttccgctttccagcgtggaagaggatgccttctgagggttgacgccattgtcataaacggtgaacttggtgcccatcaggttggaccgcaatttcccgatatagctatcgcctccccgagacaagtctgttgggtccacagagatgaggtaattggaagttttactcttctttctcttcctgcccgccaggaggaacaccttcttgccatcctcacggtctaggtgcagaaagtaggtggggtacatgccgcggtccatccccttcttgtc 209|ccgagtgatgcggcatttgatggtgatcccttgtggggctggcctcagtgcaaactcctctagatcctggacctcaatatccacctcgagagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttat 209|ttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaa 209|gttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P23Y [UBQ10prom::CITRINE-2xPH(OSBP)/pB7m34GW] 27|0 222|3 33|11700 236|628939877 237|628939877 26|2542 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9683 56|9703 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8933 56|8953 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11693 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 2xPH domain OSBP human/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|KGAGAGAGS linker 53|1 55|8592 56|8618 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain OSBP human 53|1 55|8619 56|8924 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain OSBP human 53|1 55|8280 56|8585 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8958 56|9674 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9705 56|10204 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10009 56|10093 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9705 56|10008 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|10094 56|11690 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xPH domain OSBP human/pDONRP2R-P3 72|0 73|0 74|1 75|678 76|8263 77|8940 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8941 77|9690 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9691 77|11700 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatgccagcatcttcacagctttggccttggccagttccagggccgtcacccagcgctgccgctcaacttctgaactagctttcagatggtaggtctgagcacccccattggaaatgatgaag 209|ttgcaggagtcctccacggtgatgttggctgtggcgaggttgatggtaccacggcaggtatgtctcatttctgcctttgatctgtagtagctcaggagcccgttgctcagcacgaaccatcgccgctggtagcctttgatataattggtccatttgaagagccagccctctcgagccgagcccgaagatctagaaccagcaccagcaccagcaccctttgccagcatcttcacagctttggccttggccagttccagggccgtcacccagcgctgccgctcaacttctgaactagctttcagatggtaggtctgagcacccccattggaaatgatgaagttgcaggagtcctccacggtgatgttggctgtggcgaggttgatggtaccacggcaggtatgtctcatttctgcctttgatctgtagtagctcaggagcccgttgctcagcacgaaccatcgccgctggtagcctttgatataattggtccatttgaagagccagccctctcgagccgagcccgaagatctagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaa 209|ttattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgg 209|gacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P1R [UBQ10prom::2xCHERRY-1xFYVE(HRS)/pH7m34GW] 27|0 222|3 33|12745 236|616256523 237|616256523 26|2457 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10728 56|10748 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9153 56|9173 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12738 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCHERRY-1xmyc/pDONR221, 1xFYVE wSTOP/pDONRP2R-P3 and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|FYVE 53|1 55|8920 56|9150 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|9244 56|9948 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|9982 56|10686 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9949 56|9972 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|10687 56|10710 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY 53|1 55|9244 56|10716 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|9181 56|9210 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|9220 56|9234 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-1xMyc 53|1 55|9181 56|10716 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10750 56|11249 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11054 56|11138 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10750 56|11053 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11139 56|12735 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xFYVE wSTOP/pDONRP2R-P3 72|0 73|0 74|1 75|258 76|8903 77|9160 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCHERRY-1xmyc/pDONR221 72|0 73|0 74|1 75|1575 76|9161 77|10735 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10736 77|12745 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatgccttcttgttcagctgctcatagcagggctcacacacgcgcacctccttctcaatgccgaacttggggatggtggagtacttggaggagcacttgccacagaagatctgcccacatgctcggcagtgatgcttgcgggtcaccaccccaaactgtactctgcaccgatggcattcctcagcatccacccagtcaggggctctttcagcagcgaacatggcatcactttcagccactttgtacaagaaagctgggtaattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttca 209|gcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaa 209|ggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggatttta 209|tgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P15R [UBQ10prom::2xCHERRY-1xTUBBY-c(TUBBY)y/pH7m34GW] 27|0 222|3 33|13303 236|616256442 237|616256442 26|2455 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|11286 56|11306 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9711 56|9731 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|13296 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCHERRY-1xmyc/pDONR221, Tubby/pDONRP2R-P3 (p3') and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|Tubby domain 53|1 55|8920 56|9708 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|9802 56|10506 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|10540 56|11244 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10507 56|10530 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|11245 56|11268 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY 53|1 55|9802 56|11274 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|9739 56|9768 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|9778 56|9792 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-1xMyc 53|1 55|9739 56|11274 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|11308 56|11807 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11612 56|11696 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|11308 56|11611 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11697 56|13293 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|Tubby/pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|816 76|8903 77|9718 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCHERRY-1xmyc/pDONR221 72|0 73|0 74|1 75|1575 76|9719 77|11293 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|11294 77|13303 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactcgcaggccagcttgctgtcaaagctggacagagcaatggcaaaggcctgcagtgcacacagtgggtagttgtaatccatggtgaacacatcttctgctacccggccaaactgcatgacgatgtagtccgggtcattgccgtggatgatctggaagttcttcacagaagcctgtgtgacacggccgtggaagttaagtacataggactgtgtgtcatcattccagactggcgtcttgttctgcagctcaatgatgctctccgtgttcttgttctgccagcgtgctaacagggtctcatgttcattgcgggggcggatacagactctctcatgaaccatgttcatgcctgggacgatcacactcatcttccgaggtcccttgaagcctaggacatttgtctcatagcacaccgctgccagctcctggcgcaaggttccgctttccagcgtggaagaggatgccttctgagggttgacgccattgtcataaacggtgaacttggtgcccatcaggttggaccgcaatttcccgatatagctatcgcctccccgagacaagtctgttgggtccacagagatgaggtaattggaagttttactcttctttctcttcctgcccgccaggaggaacaccttcttgccatcctcacggtctaggtgcagaaagtaggtggggtacatgccgcggtccatccccttcttgtcccgagtgatgcggcatttgatggtgatcccttgtggggctggcctcagtgcaaactcctctagatcctggacctcaatatccacagccactttgtacaagaaagctgggtaattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtc 209|ccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatg 209|aactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaaca 209|agaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P18C [UBQ10prom::2xCyPet-2xFYVE(HRS)/pK7m34GW] 27|0 222|3 33|12760 236|582631479 237|582631479 26|1908 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10743 56|10763 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9012 56|9032 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12753 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221, 2XFYVE (+STOP)/pDONRP2R-P3 and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|6 52|FYVE 53|1 55|8779 56|9009 57|1 281|1 282|1 283|1 284|1 285|3363..3593 50 45 51|6 52|FYVE 53|1 55|8533 56|8763 57|1 281|1 282|1 283|1 284|1 285|3609..3839 50 45 51|21 52|cyPET 53|1 55|9940 56|10680 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|9202 56|9942 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|9154 56|9171 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|9106 56|9126 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|10660 56|10689 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9922 56|9942 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9184 56|9198 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10765 56|11264 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11069 56|11153 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10765 56|11068 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11154 56|12750 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2XFYVE (+STOP)/pDONRP2R-P3 72|0 73|0 74|1 75|504 76|8516 77|9019 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221 72|0 73|0 74|1 75|1731 76|9020 77|10750 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10751 77|12760 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgctagtgccttcttgttcagctgctcatagcagggctcacacacgcgcacctccttctcaatgccgaacttggggatggtggagtacttggaggagcacttgccacagaagatctgcccacatgctcggcagtgatgcttgcgggtcaccaccccaaactgtactctgcaccgatggcattcctcagcatccacccagtcaggggctctttcagcagcgaacatggcatcactttcggatccttgtccttgtgccttcttgttcagctgctcatagcagggctcacacacgcgcacctccttctcaatgccgaacttggggatggtggagtacttggaggagcacttgccacagaagatctgcccacatgctcggcagtgatgcttgcgggtcaccaccccaaactgtactctgcaccgatggcattcctcagcatccacccagtcaggggctctttcagcagcgaacatggcatcactttcagccactttgtacaagaaagctgggtagtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagct 209|tgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactagg 209|tcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagca 209|agtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P21C [UBQ10prom::2xCyPet-2xPH(FAPP1)/pK7m34GW] 27|0 222|3 33|12928 236|582631466 237|582631466 26|1907 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10911 56|10931 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9180 56|9200 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12921 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221, 2xPH FAPP1 human/pDONRP2R-P3 and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|15 52|KGAGAGAGS linker 53|1 55|8842 56|8868 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain FAPP1 53|1 55|8869 56|9171 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain FAPP1 53|1 55|8533 56|8835 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|1 55|10108 56|10848 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|9370 56|10110 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|9322 56|9339 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|9274 56|9294 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|10828 56|10857 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10090 56|10110 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9352 56|9366 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10933 56|11432 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11237 56|11321 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10933 56|11236 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11322 56|12918 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xPH FAPP1 human/pDONRP2R-P3 72|0 73|0 74|1 75|672 76|8516 77|9187 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221 72|0 73|0 74|1 75|1731 76|9188 77|10918 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10919 77|12928 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatttagtccttgtatcagtcaaacatgctttggagctccccagagcgaccagccacctctgtctttcagctgcattcactgccttcatgtagaaatgctgctctccaggaatgattaattccattcttgtgttgtctgctgaatgaactttaatttcacaaactgccatctttatgcttcctttgctccctttgcaaacatcatcttgtgaatcatagtaggataagattccattatctaaaacaaaccaacgaggctgccagcctgtgagatagttggtccacttgtacaacaccccctccatctcgagagaaccagcaccagcaccagcaccctttttagtccttgtatcagtcaaacatgctttggagctccccagagcgaccagccacctctgtctttcagctgcattcactgccttcatgtagaaatgctgctctccaggaatgattaattccattcttgtgttgtctgctgaatgaactttaatttcacaaactgccatctttatgcttcctttgctccctttgcaaacatcatcttgtgaatcatagtaggataagattccattatctaaaacaaaccaacgaggctgccagcctgtgagatagttggtccacttgtacaacaccccctccatctcgagagccactttgtacaagaaagctgggtagtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatgg 209|tccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatc 209|gaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctgg 209|aagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P24C [UBQ10prom::2xCyPet-2xPH(PLC)/pK7m34GW] 27|0 222|3 33|13351 236|582631452 237|582631452 26|1906 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|11334 56|11354 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9603 56|9623 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8508 56|8528 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|13344 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221, 2xPLCd1 PH domain Rat/pDONRP2R-P3 and Destination Vector pK7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Kan 53|1 55|6771 56|8167 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8201 56|8426 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8484 56|8499 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|2xPH domain PLCd1 53|1 55|8533 56|9600 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain PLCd1 Rat 53|1 55|9076 56|9600 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain PLCd1 Rat 53|1 55|8533 56|9044 57|1 281|1 282|1 283|1 284|1 50 45 51|84 52|GAGARS linker 53|1 55|9045 56|9075 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|cyPET 53|1 55|10531 56|11271 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|CyPET 53|1 55|9793 56|10533 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|6xHIS 53|1 55|9745 56|9762 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|TEV cleavage site 53|1 55|9697 56|9717 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PolyA linker 53|1 55|11251 56|11280 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10513 56|10533 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|9775 56|9789 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|11356 56|11855 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11660 56|11744 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|11356 56|11659 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11745 56|13341 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pK7m34GW,0 72|0 73|0 74|1 75|8515 76|1 77|8515 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xPLCd1 PH domain Rat/pDONRP2R-P3 72|0 73|0 74|1 75|1095 76|8516 77|9610 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCyPET-Hellfire wKOZAKnoSTOP/pDONR221 72|0 73|0 74|1 75|1731 76|9611 77|11341 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|11342 77|13351 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggt 209|gatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactggatgttgagctccttcaggaagtccttcagctccttgaagttcatcttgttgtccttgtttttatcagcctttcgcaagcaggagtgaatccagtgctgcagcttctgccgctggtccatggagccggagtggtggatgatcttgcgcaggccctgcacccagtgctgagcgtcagctggtgatggggcaatgaggtctagggtgttgcgctggtccttgaagacaatggagaagcatcgatcctcggggatgtctcgggcaaacttctccaggccttctgtgcggtgtcccatccgtacctcctgaatgtcctcgatggagaacagctgcgactccggggacctcatgacctttcgagattcctgccagatggtcttgcagtcctcctgtagcttgtagaagcgttccctacgccacgagctggacttcaccttcagaagctggctgcccttcagaagggcctgaaggtccgggtcatcctggagcccgtgcagggtcaggaagtccctacccgagtccatggatctagctcctgcaccctggatgttgagctccttcaggaagtccttcagctccttgaagttcatcttgttgtccttgtttttatcagcctttcgcaagcaggagtgaatccagtgctgcagcttctgccgctggtccatggagccggagtggtggatgatcttgcgcaggccctgcacccagtgctgagcgtcagctggtgatggggcaatgaggtctagggtgttgcgctggtccttgaagacaatggagaagcatcgatcctcggggatgtctcgggcaaacttctccaggccttctgtgcggtgtcccatccgtacctcctgaatgtcctcgatggagaacagctgcgactccggggacctcatgacctttcgagattcctgccagatggtcttgcagtcctcctgtagcttgtagaagcgttccctacgccacgagctggacttcaccttcagaagctggctgcccttcagaagggcctgaaggtccgggtcatcctggagcccgtgcagggtcaggaagtcccggcccgagtccatagccactttgtacaagaaagctgggtagtcatcgtcatccttgtagtcgatgtcatgatctttataatcaccgtcatggtctttgtagtcgagctcaccctgaaaatacaaattctcgctagcagtagttggaatatcataatcgtgatggtgatggtgatgcgatcctcttccagcagcagcagcagcttctagtcctt 209|tgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagctccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagtcctttgtacagttcgtccatgccgtgggtgatgccggcggcggtcacgaactccagcagcaccatgtggtcccgcttctcgttggggtccttgctcagggcgctctgggtgctcaggtagtggttgtcgggcaggatcacggggccgtcgccgatgggggtgttctgctggtagtggtcggccagctgcacgctgccgtcggtgatgttgtgccgggccttgaagttggccttgatgccgttcttctgcttgtcggcggtgatgtacacgttgtggctgatgtagttgtactccagcttgtggcccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatccggttcaccagggtgtcgccctcgaacttcacctcggcccgggtcttgtagttgccgtcgtccttgaagaagatggtccgctcctgcacgtagccctcgggcatcacgctcttgaagaagtcgtgctgcttcatgtggtcggggtaccggctgaagcactgcacgccccaggtcagggtggtcaccagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcgtcgccctcgccctcgccgctcacgctgaacttgtggccgttcacgtcgccctccagc 209|tccaccaggatgggcacgatgccgccgaacagttcctctcccttgctcacagcggcagcagcagcatctagaggatccaagcttatcgatttcgaacccggggtaccgaattcctcgagcatcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcc 209|tagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P18R [UBQ10prom::2xCHERRY-2xFYVE(HRS)/pH7m34GW] 27|0 222|3 33|12991 236|582631332 237|582631332 26|1904 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|10974 56|10994 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9399 56|9419 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|12984 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCHERRY-1xmyc/pDONR221, 2XFYVE (+STOP)/pDONRP2R-P3 and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|6 52|FYVE 53|1 55|9166 56|9396 57|1 281|1 282|1 283|1 284|1 285|3363..3593 50 45 51|6 52|FYVE 53|1 55|8920 56|9150 57|1 281|1 282|1 283|1 284|1 285|3609..3839 50 45 51|21 52|mCHERRY 53|1 55|9490 56|10194 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|10228 56|10932 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10195 56|10218 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|10933 56|10956 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY 53|1 55|9490 56|10962 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|9427 56|9456 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|9466 56|9480 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-1xMyc 53|1 55|9427 56|10962 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|10996 56|11495 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11300 56|11384 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|10996 56|11299 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11385 56|12981 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2XFYVE (+STOP)/pDONRP2R-P3 72|0 73|0 74|1 75|504 76|8903 77|9406 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCHERRY-1xmyc/pDONR221 72|0 73|0 74|1 75|1575 76|9407 77|10981 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|10982 77|12991 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgctagtgccttcttgttcagctgctcatagcagggctcacacacgcgcacctccttctcaatgccgaacttggggatggtggagtacttggaggagcacttgccacagaagatctgcccacatgctcggcagtgatgcttgcgggtcaccaccccaaactgtactctgcaccgatggcattcctcagcatccacccagtcaggggctctttcagcagcgaacatggcatcactttcggatccttgtccttgtgccttcttgttcagctgctcatagcagggctcacacacgcgcacctccttctcaatgccgaacttggggatggtggagtacttggaggagcacttgccacagaagatctgcccacatgctcggcagtgatgcttgcgggtcaccaccccaaactgtactctgcaccgatggcattcctcagcatccacccagtcaggggctctttcagcagcgaacatggcatcactttcagccactttgtacaagaaagctgggtaattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagt 209|tggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatc 209|taagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcg 209|atcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P21R [UBQ10prom::2xCHERRY-2xPH(FAPP1)/pH7m34GW] 27|0 222|3 33|13159 236|582631322 237|582631322 26|1903 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|11142 56|11162 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9567 56|9587 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|13152 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCHERRY-1xmyc/pDONR221, 2xPH FAPP1 human/pDONRP2R-P3 and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|15 52|KGAGAGAGS linker 53|1 55|9229 56|9255 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain FAPP1 53|1 55|9256 56|9558 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain FAPP1 53|1 55|8920 56|9222 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|9658 56|10362 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|10396 56|11100 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10363 56|10386 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|11101 56|11124 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY 53|1 55|9658 56|11130 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|9595 56|9624 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|9634 56|9648 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-1xMyc 53|1 55|9595 56|11130 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|11164 56|11663 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11468 56|11552 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|11164 56|11467 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11553 56|13149 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xPH FAPP1 human/pDONRP2R-P3 72|0 73|0 74|1 75|672 76|8903 77|9574 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCHERRY-1xmyc/pDONR221 72|0 73|0 74|1 75|1575 76|9575 77|11149 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|11150 77|13159 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatttagtccttgtatcagtcaaacatgctttggagctccccagagcgaccagccacctctgtctttcagctgcattcactgccttcatgtagaaatgctgctctccaggaatgattaattccattcttgtgttgtctgctgaatgaactttaatttcacaaactgccatctttatgcttcctttgctccctttgcaaacatcatcttgtgaatcatagtaggataagattccattatctaaaacaaaccaacgaggctgccagcctgtgagatagttggtccacttgtacaacaccccctccatctcgagagaaccagcaccagcaccagcaccctttttagtccttgtatcagtcaaacatgctttggagctccccagagcgaccagccacctctgtctttcagctgcattcactgccttcatgtagaaatgctgctctccaggaatgattaattccattcttgtgttgtctgctgaatgaactttaatttcacaaactgccatctttatgcttcctttgctccctttgcaaacatcatcttgtgaatcatagtaggataagattccattatctaaaacaaaccaacgaggctgccagcctgtgagatagttggtccacttgtacaacaccccctccatctcgagagccactttgtacaagaaagctgggtaattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttct 209|tggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattc 209|gatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatga 209|gtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P24R [UBQ10prom::2xCHERRY-2xPH(PLC)/pH7m34GW] 27|0 222|3 33|13582 236|582631311 237|582631311 26|1902 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|11565 56|11585 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9990 56|10010 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8895 56|8915 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|13575 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 2xCHERRY-1xmyc/pDONR221, 2xPLCd1 PH domain Rat/pDONRP2R-P3 and Destination Vector pH7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Hyg 53|1 55|6771 56|8557 57|1 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|8588 56|8813 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8871 56|8886 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|2xPH domain PLCd1 53|1 55|8920 56|9987 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain PLCd1 Rat 53|1 55|9463 56|9987 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain PLCd1 Rat 53|1 55|8920 56|9431 57|1 281|1 282|1 283|1 284|1 50 45 51|84 52|GAGARS linker 53|1 55|9432 56|9462 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|10081 56|10785 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|1 55|10819 56|11523 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|1 55|10786 56|10809 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|11524 56|11547 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY 53|1 55|10081 56|11553 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|1 55|10018 56|10047 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|polyAlinker 53|1 55|10057 56|10071 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|2xmCHERRY-1xMyc 53|1 55|10018 56|11553 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|11587 56|12086 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|11891 56|11975 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|11587 56|11890 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|11976 56|13572 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pH7m34GW,0 72|0 73|0 74|1 75|8902 76|1 77|8902 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xPLCd1 PH domain Rat/pDONRP2R-P3 72|0 73|0 74|1 75|1095 76|8903 77|9997 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|2xCHERRY-1xmyc/pDONR221 72|0 73|0 74|1 75|1575 76|9998 77|11572 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|11573 77|13582 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacagg 209|attcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtgatacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaata 209|atggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactggatgttgagctccttcaggaagtccttcagctccttgaagttcatcttgttgtccttgtttttatcagcctttcgcaagcaggagtgaatccagtgctgcagcttctgccgctggtccatggagccggagtggtggatgatcttgcgcaggccctgcacccagtgctgagcgtcagctggtgatggggcaatgaggtctagggtgttgcgctggtccttgaagacaatggagaagcatcgatcctcggggatgtctcgggcaaacttctccaggccttctgtgcggtgtcccatccgtacctcctgaatgtcctcgatggagaacagctgcgactccggggacctcatgacctttcgagattcctgccagatggtcttgcagtcctcctgtagcttgtagaagcgttccctacgccacgagctggacttcaccttcagaagctggctgcccttcagaagggcctgaaggtccgggtcatcctggagcccgtgcagggtcaggaagtccctacccgagtccatggatctagctcctgcaccctggatgttgagctccttcaggaagtccttcagctccttgaagttcatcttgttgtccttgtttttatcagcctttcgcaagcaggagtgaatccagtgctgcagcttctgccgctggtccatggagccggagtggtggatgatcttgcgcaggccctgcacccagtgctgagcgtcagctggtgatggggcaatgaggtctagggtgttgcgctggtccttgaagacaatggagaagcatcgatcctcggggatgtctcgggcaaacttctccaggccttctgtgcggtgtcccatccgtacctcctgaatgtcctcgatggagaacagctgcgact 209|ccggggacctcatgacctttcgagattcctgccagatggtcttgcagtcctcctgtagcttgtagaagcgttccctacgccacgagctggacttcaccttcagaagctggctgcccttcagaagggcctgaaggtccgggtcatcctggagcccgtgcagggtcaggaagtcccggcccgagtccatagccactttgtacaagaaagctgggtaattcaagtcctcttcagaaatgagcttttgctccatagctttagcagcagcagcagcttctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcaccacgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagtcccttgtacagctcgtccatgccgccggtggagtggcggccctcggcgcgttcgtactgttccacgatggtgtagtcctcgttgtgggaggtgatgtccaacttgatgttgacgttgtaggcgccgggcagctgcacgggcttcttggccttgtaggtggtcttgacctcagcgtcgtagtggccgccgtccttcagcttcagcctctgcttgatctcgcccttcagggcgccgtcctcggggtacatccgctcggaggaggcctcccagcccatggtcttcttctgcattacggggccgtcggaggggaagttggtgccgcgcagcttcaccttgtagatgaactcgccgtcctgcagggaggagtcctgggtcacggtcacc 209|acgccgccgtcctcgaagttcatcacgcgctcccacttgaagccctcggggaaggacagcttcaagtagtcggggatgtcggcggggtgcttcacgtaggccttggagccgtacatgaactgaggggacaggatgtcccaggcgaagggcagggggccacccttggtcaccttcagcttggcggtctgggtgccctcgtaggggcggccctcgccctcgccctcgatctcgaactcgtggccgttcacggagccctccatgtgcaccttgaagcgcatgaactccttgatgatggccatgttatcctcctcgcccttgctcacagcggcagcggcagcaggcaaagggtctagcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgta 209|tcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P6Y [UBQ10prom::CITRINE-1xGRAM9Atg26)/pB7m34GW] 27|0 222|3 33|11250 236|574874387 237|574874387 26|1720 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9233 56|9253 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8483 56|8503 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11243 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 1xGRAM domain/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|GRAM domain 53|1 55|8280 56|8480 57|1 281|1 282|1 283|1 284|1 50 45 51|62 52|Y57P 53|1 54|Y57P when the conserved Tyr57 is mutated into a Proline, the GRAM domain lose its ability to bind PI4P 55|8310 56|8312 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8508 56|9224 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9255 56|9754 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9559 56|9643 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9255 56|9558 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9644 56|11240 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xGRAM domain/pDONRP2R-P3 72|0 73|0 74|1 75|228 76|8263 77|8490 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8491 77|9240 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9241 77|11250 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctacttcaaattataaacattctcaatatcggagtatggaagtatcatagttgtgtttgtgccgggaaataaagatctgtagcaaatgcaagttgttcctaggtatactttgccgtatacaggt 209|atatttttttgaaaatggcagaagtaactagccagtagctcctctgaatcaggaagggaaaaatgctttcgaaatctggaagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttg 209|aggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattct 209|attgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P5Y [UBQ10prom::CITRINE-1xPH(FAPP1)/pB7m34GW] 27|0 222|3 33|11358 236|574874261 237|574874261 26|1718 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9341 56|9361 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8591 56|8611 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11351 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 1xPH FAPP1 human/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PH domain FAPP1 53|1 55|8280 56|8582 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8616 56|9332 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9363 56|9862 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9667 56|9751 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9363 56|9666 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9752 56|11348 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xPH FAPP1 human/pDONRP2R-P3 72|0 73|0 74|1 75|336 76|8263 77|8598 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8599 77|9348 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9349 77|11358 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatttagtccttgtatcagtcaaacatgctttggagctccccagagcgaccagccacctctgtctttcagctgcattcactgccttcatgtagaaatgctgctctccaggaatgattaattcc 209|attcttgtgttgtctgctgaatgaactttaatttcacaaactgccatctttatgcttcctttgctccctttgcaaacatcatcttgtgaatcatagtaggataagattccattatctaaaacaaaccaacgaggctgccagcctgtgagatagttggtccacttgtacaacaccccctccatctcgagagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtc 209|acgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaag 209|tttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P14Y [UBQ10prom::CITRINE-1xPH(PLC)/pB7m34GW] 27|0 222|3 33|11574 236|574874234 237|574874234 26|1717 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9557 56|9577 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8807 56|8827 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11567 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 1x PLCd1 (PH domain rat)/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|PH domain PLCd1 (Rat) 53|1 55|8280 56|8804 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8832 56|9548 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9579 56|10078 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9883 56|9967 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9579 56|9882 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9968 56|11564 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1x PLCd1 (PH domain rat)/pDONRP2R-P3 72|0 73|0 74|1 75|552 76|8263 77|8814 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8815 77|9564 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9565 77|11574 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactggatgttgagctccttcaggaagtccttcagctccttgaagttcatcttgttgtccttgtttttatcagcctttcgcaagcaggagtgaatccagtgctgcagcttctgccgctggtcc 209|atggagccggagtggtggatgatcttgcgcaggccctgcacccagtgctgagcgtcagctggtgatggggcaatgaggtctagggtgttgcgctggtccttgaagacaatggagaagcatcgatcctcggggatgtctcgggcaaacttctccaggccttctgtgcggtgtcccatccgtacctcctgaatgtcctcgatggagaacagctgcgactccggggacctcatgacctttcgagattcctgccagatggtcttgcagtcctcctgtagcttgtagaagcgttccctacgccacgagctggacttcaccttcagaagctggctgcccttcagaagggcctgaaggtccgggtcatcctggagcccgtgcagggtcaggaagtcccggcccgagtccatagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatct 209|aaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtg 209|ttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P1Y [UBQ10prom::CITRINE-1xFYVE(HRS)/pB7m34GW] 27|0 222|3 33|11280 236|574874202 237|574874202 26|1716 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9263 56|9283 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8513 56|8533 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11273 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 1xFYVE wSTOP/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|FYVE 53|1 55|8280 56|8510 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8538 56|9254 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9285 56|9784 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9589 56|9673 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9285 56|9588 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9674 56|11270 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xFYVE wSTOP/pDONRP2R-P3 72|0 73|0 74|1 75|258 76|8263 77|8520 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8521 77|9270 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9271 77|11280 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctatgccttcttgttcagctgctcatagcagggctcacacacgcgcacctccttctcaatgccgaacttggggatggtggagtacttggaggagcacttgccacagaagatctgcccacatgct 209|cggcagtgatgcttgcgggtcaccaccccaaactgtactctgcaccgatggcattcctcagcatccacccagtcaggggctctttcagcagcgaacatggcatcactttcagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttg 209|tttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttct 209|tttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P2Y [UBQ10prom::CITRINE-1xAtPH(PH1)/pB7m34GW] 27|0 222|3 33|11373 236|574874177 237|574874177 26|1715 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9356 56|9376 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8606 56|8626 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11366 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 1xPH domain AtPH1/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|PH domain AtPH1 53|1 55|8280 56|8597 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8631 56|9347 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9378 56|9877 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9682 56|9766 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9378 56|9681 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9767 56|11363 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xPH domain AtPH1/pDONRP2R-P3 72|0 73|0 74|1 75|351 76|8263 77|8613 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8614 77|9363 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9364 77|11373 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctattgcacaatcgatcttccaatcgaattaatccactcttctttctccttctcattatcagcaatgaagaacatggtatagctaccactagatagctcaaaagcaaaaggcttattcacaaca 209|tcctcagctcctttcacggtgagacaatcaccaacggagatcacaccacgcggcgtagatccacgaattccagcagcggcttgatctttgaaccagagaagcttccctcgtttgagaacgaaccaacgacgacgccaggttttgatgtaatcgccttgctttgtgagccaaccagaacgctcagggtttgaccagaactcgagagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagac 209|gaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaag 209|tatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P11Y [UBQ10prom::CITRINE-1xPH(TAPP2)/pB7m34GW] 27|0 222|3 33|11424 236|574863417 237|574863417 26|1712 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9407 56|9427 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8657 56|8677 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11417 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 1xPH domain TAPP2 mouse with GAGA linker/pDONRP2R-P3 (p3') and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|Cter PH domain TAPP2 mouse 53|1 55|8280 56|8624 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8682 56|9398 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9429 56|9928 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9733 56|9817 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9429 56|9732 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9818 56|11414 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xPH domain TAPP2 mouse with GAGA linker/pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|402 76|8263 77|8664 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8665 77|9414 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9415 77|11424 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctagtggcacttcagagcctggacagctgctccaatcccctcgatccagctgtgcatgtcctcagggctgtccgcctgtacgtagaacgtcctggaggtggttatgatttcaaacaggttgtcc 209|ctcattaagagatcaccagacttgaccagacactcgtgagtcttgagaacatccttgagcggtatggtacgcagaggctctctgtcctgctcacacttgaagtagcagatggtaaagtcatcgagggcaaagaagcgtcgtttccaactcttccgcacattcccttgcttcacacagtagccactcttaatgaggggaggcccagtggaagggcggcagcctgaagaaccagcaccagcaccagcaccctcgagagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtga 209|agaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggtt 209|acatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P10Y [UBQ10prom::mCITRINE-1xPH(TAPP1)/pB7m34GW] 27|0 222|3 33|11451 236|574863386 237|574863386 26|1711 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9434 56|9454 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8684 56|8704 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11444 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 1xPH domain TAPP1 with GAGA linker/pDONRP2R-P3 (p3'1x) and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|Cter PH domain TAPP1 human 53|1 55|8280 56|8651 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|8709 56|9425 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9456 56|9955 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9760 56|9844 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9456 56|9759 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9845 56|11441 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xPH domain TAPP1 with GAGA linker/pDONRP2R-P3 (p3'1x) 72|0 73|0 74|1 75|429 76|8263 77|8691 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|8692 77|9441 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9442 77|11451 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctaaccggggggatgctcagaagacgcagatctgccgggaccccgctgtgctacaatggcgccagagactgctttaatccaactgtgcatctcttcagggctatcagcctgcacatagaaagtt 209|cgagacgttgttacaatttcaaagaggttgtccctcatcattatgtcgctttgcttacattcctggactttatgaacctctttaagtggtattacacgaagaggttccttttccagttcagatttgaagtagcctattgtgttttcatccaattgaaaatatcttctcttccagtttttcatcactgctccttgttttacacaatatccagctttgataaccgcactatcttgaggtggtttaggagtaaaagaaccagcaccagcaccagcaccctcgagagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagat 209|tttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtg 209|gattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P17Y [UBQ10prom::1xPH(BTK)-CITRINE/pB7m34GW] 27|0 222|3 33|11481 236|574863060 237|574863060 26|1709 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9464 56|9484 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8999 56|9019 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11474 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, 1xPH BTK GAGAnoSTOP/pDONR221, mCITRINE+STOP/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|mCITRINE 53|1 55|8280 56|8996 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|GAGA linker 53|1 55|9033 56|9053 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH doamin BTK 53|1 55|9054 56|9455 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9486 56|9985 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9790 56|9874 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9486 56|9789 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9875 56|11471 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINE+STOP/pDONRP2R-P3 72|0 73|0 74|1 75|744 76|8263 77|9006 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|1xPH BTK GAGAnoSTOP/pDONR221 72|0 73|0 74|1 75|465 76|9007 77|9471 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9472 77|11481 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcttacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtgg 209|ttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatagccactttgtacaagaaagctgggtactcgagagaaccagcaccagcaccagcaccgtaccggattacgtttttgagctggtgaatccaccgcttccttagttcttcagttggggagaagacgtagagaggcccttcatcatatacaacctggaagggataagggaacctttcaatgattgaaatttgctccatttcactggactcttcacctcttctcggaatctgtctttctggaggaggatttttttcaggaaccactgtttcaacacaagtgatcttctcaacatctattgaacccttcttactgcctcttctcccacgttcaaagtcatactcatagtaggagagtttgtgcacggtcaagagaaacaggcgcttcttgaagtttagaggtgatgttttctttttctgttgggatcgcttcagaaagatgctctccagaatcactgcggccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacgg 209|tagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccga 209|ttcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P8Y [UBQ10prom::CITRINE-3xPHD(ING2)/pB7m34GW] 27|0 222|3 33|11838 236|569948630 237|569948630 26|1655 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9821 56|9841 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9071 56|9091 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11831 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, 3xPHD ING2 wSTOP/pDONRP2R-P3 (p3') and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|PHD ING2 53|1 55|8802 56|9048 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PHD ING2 53|1 55|8553 56|8798 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PHD ING2 53|1 55|8307 56|8549 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|xPHD ING2 53|1 55|8307 56|9048 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|9096 56|9812 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9843 56|10342 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10147 56|10231 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9843 56|10146 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|10232 56|11828 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|3xPHD ING2 wSTOP/pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|816 76|8263 77|9078 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|9079 77|9828 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9829 77|11838 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgccgggcccgcggtaccgtcgactcatcactacctcgctcttctctccttttttgtcttttcggtacttttgtccatggttttctcattgtctcccctacactttgggcaataccatttccccttg 209|ggtttataggtgagtgaaacacatgaaaagtgaaaccattcaatgggacactgttcattgtcacagcctatcatctccccgtaggacacttggttacacaagcagtaggtgggctcattgggatcgatggcaaactcgacaggggatgcgaattccctcgctcttctctccttttttgtcttttcggtacttttgtccatggttttctcattgtctcccctacactttgggcaataccatttccccttgggtttataggtgagtgaaacacatgaaaagtgaaaccattcaatgggacactgttcattgtcacagcctatcatctccccgtaggacacttggttacacaagcagtaggtgggctcattgggatcgatggcaaactcgacaggggatgcgaattccctcgctcttctctccttttttgtcttttcggtacttttgtccatggttttctcattgtctcccctacactttgggcaataccatttccccttgggtttataggtgagtgaaacacatgaaaagtgaaaccattcaatgggacactgttcattgtcacagcctatcatctccccgtaggacacttggttacacaagcagtaggtgggctcattgggatcgatggcaaactcgacaggggatgccgagatctgagtccggccggaagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcc 209|cttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaaga 209|acagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P15Y [UBQ10prom::CITRINE-TUBBY-C(TUBBY)/pB7m34GW] 27|0 222|3 33|11838 236|569948563 237|569948563 26|1653 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9821 56|9841 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|9071 56|9091 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11831 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, mCITRINEnoSTOP/pDONR221, Tubby/pDONRP2R-P3 (p3') and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|21 52|Tubby domain 53|1 55|8280 56|9068 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|1 55|9096 56|9812 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9843 56|10342 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|10147 56|10231 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9843 56|10146 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|10232 56|11828 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|Tubby/pDONRP2R-P3 (p3') 72|0 73|0 74|1 75|816 76|8263 77|9078 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINEnoSTOP/pDONR221 72|0 73|0 74|1 75|750 76|9079 77|9828 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9829 77|11838 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcctactcgcaggccagcttgctgtcaaagctggacagagcaatggcaaaggcctgcagtgcacacagtgggtagttgtaatccatggtgaacacatcttctgctacccggccaaactgcatgacg 209|atgtagtccgggtcattgccgtggatgatctggaagttcttcacagaagcctgtgtgacacggccgtggaagttaagtacataggactgtgtgtcatcattccagactggcgtcttgttctgcagctcaatgatgctctccgtgttcttgttctgccagcgtgctaacagggtctcatgttcattgcgggggcggatacagactctctcatgaaccatgttcatgcctgggacgatcacactcatcttccgaggtcccttgaagcctaggacatttgtctcatagcacaccgctgccagctcctggcgcaaggttccgctttccagcgtggaagaggatgccttctgagggttgacgccattgtcataaacggtgaacttggtgcccatcaggttggaccgcaatttcccgatatagctatcgcctccccgagacaagtctgttgggtccacagagatgaggtaattggaagttttactcttctttctcttcctgcccgccaggaggaacaccttcttgccatcctcacggtctaggtgcagaaagtaggtggggtacatgccgcggtccatccccttcttgtcccgagtgatgcggcatttgatggtgatcccttgtggggctggcctcagtgcaaactcctctagatcctggacctcaatatccacagccactttgtacaagaaagctgggtacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtggttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcc 209|cttgctcaccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaaga 209|acagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P12Y [UBQ10prom::ENTH(Ent3p)-CITRINE/pB7m34GW] 27|0 222|3 33|11589 236|569948333 237|569948333 26|1652 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9572 56|9592 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8999 56|9019 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11582 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, ENTH domain Ent3p noSTOP/pDONR221, mCITRINE+STOP/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|mCITRINE 53|1 55|8280 56|8996 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|ENTH domain Ent3 53|1 55|9024 56|9563 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9594 56|10093 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9898 56|9982 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9594 56|9897 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9983 56|11579 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINE+STOP/pDONRP2R-P3 72|0 73|0 74|1 75|744 76|8263 77|9006 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|ENTH domain Ent3p noSTOP/pDONR221 72|0 73|0 74|1 75|573 76|9007 77|9579 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9580 77|11589 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcttacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtgg 209|ttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatagccactttgtacaagaaagctgggtaactgccatcagcagaggcagatcctccagcaacgcccttgtacttctttgccgtttctcttgccttttttctttctgcgcgtatcttattatcgtcgctcaataattcaatcaaagctttgaccctagttctgacattaattccttgatctcttccctgggaatcaatatagtgaaaagtttctaaaatcctaattaagttaatgctattccttgtatcatcgataaacctttcagaaccgtgtttgataaggtaatcgagcaattgcaaggctttataaatttgcctccactcactacctgccttttcagtgaaccttctgaatatcatggacaaaatttcttctctttccctgaaattgtaagttccttgagaaatctggtccattaaagtggatgaggcaccccaaggctcgttgtttgtcgcttcacgaactttcccctccatctcagtataattgaaaacaacattttgagctttacgaaaatatttcttggcgtcatataatgacatgttggctaatgtatcctctaaactcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatcta 209|agattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgat 209|catacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P9Y [UBQ10prom::1xPX(p47)-CITRINE/pB7m34GW] 27|0 222|3 33|11475 236|569948287 237|569948287 26|1651 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9458 56|9478 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8999 56|9019 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11468 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, PX p47phox-GAGAlinker noSTOP/pDONR221, mCITRINE+STOP/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|mCITRINE 53|1 55|8280 56|8996 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PX domain p47phox 53|1 55|9054 56|9446 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|GAGA linker 53|1 55|9030 56|9053 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9480 56|9979 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9784 56|9868 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9480 56|9783 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9869 56|11465 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINE+STOP/pDONRP2R-P3 72|0 73|0 74|1 75|744 76|8263 77|9006 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|PX p47phox-GAGAlinker noSTOP/pDONR221 72|0 73|0 74|1 75|459 76|9007 77|9465 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9466 77|11475 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcttacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtgg 209|ttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatagccactttgtacaagaaagctgggtactcgagagaaccagcaccagcaccagcaccctggttgtccgtggggagcttgaggtcatcagggcgcaccttgaagaagtcgaggaggtggggacagcgggagatcttggtgggcaggctcatgagcgtgctgcagtactcggtaagtgtgccctggcggttctcggcggcccgctgcccgtcaaaccacttgggagctgggaggtgggggatgatcctgttctctggattgatcgcccctgcctcaatagggaacatttcttttaaggttttatggaactcgtagatctcggtgaagcgccggtagaccaccttctccgacaggtcctgccatttcaccaggaacatgtacacatagtgctggctgggtacgaagcgcttctcaaagcccagcagggcgatgtgacggatgaaggtgtcccccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattttgagagaataaggaacacagaaatttaccttgatcacggtagaga 209|gaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaat 209|tataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P3Y [UBI10prom::1xPX(p40)-CITRINE/pB7m34GW] 27|0 222|3 33|11520 236|569948253 237|569948253 26|1650 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9503 56|9523 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8999 56|9019 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11513 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, p40phox GAGA linker noSTOP/pDONR221, mCITRINE+STOP/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|mCITRINE 53|1 55|8280 56|8996 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PX domain p40phox 53|1 55|9054 56|9494 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|GAGA linker 53|1 55|9030 56|9053 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9525 56|10024 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9829 56|9913 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9525 56|9828 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9914 56|11510 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINE+STOP/pDONRP2R-P3 72|0 73|0 74|1 75|744 76|8263 77|9006 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|p40phox GAGA linker noSTOP/pDONR221 72|0 73|0 74|1 75|504 76|9007 77|9510 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9511 77|11520 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcttacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtgg 209|ttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatagccactttgtacaagaaagctgggtactcgagagaaccagcaccagcaccagcaccgagtgcctggggcacctgctctgagtcatagggcgactggtaaaagaagatccggacgtcctcatccatcagcacccagaccggcaggctgagcaggctcttcatgtaggcgttgagggcaggtatccgcatctcggcgatctcctgtttcacacccacgtagactttggctgggagtgtgggcagggtacaggccagggcactgctcttgctgtctggcccgaagcgctcctccagcttgctctgcaaagcatggaactggcggtagcggcggtagatgaggtacttggatcctccttttgtcttcacctcgatgacgaaaacaaagtggctggtgaagcctctcttctcctcgatgtcagcaatgttggccgagatggcaacatcatccggaagctgttcaaagtcactctcggcccgcagctgctgggccacagccatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcgaagattt 209|tgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaagtagaa 209|gtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|P16Y [UBQ10prom::1xPH(AKT)-CITRINE/pB7m34GW] 27|0 222|3 33|11526 236|567895207 237|567895207 26|1587 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 34|attB1-attB2-attB3-attB4 255 224|Demo User 50 256 224|Demo User 50 1001|0 45 51|21 52|attB1 53|1 55|9509 56|9529 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB2 53|0 55|8999 56|9019 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB3 53|0 55|8255 56|8275 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attB4 53|1 55|11519 56|13 57|0 281|1 282|1 283|1 284|1 50 45 51|-1 52|Gateway Cloning Description 53|0 55|0 56|0 57|0 281|1 282|1 283|1 284|1 286|/Design_Description=Mix Entry Clones UB10-promoteur/pDONRP4-P1R, PH AKT-GAGAnoSTOP/pDONR221, mCITRINE+STOP/pDONRP2R-P3 and Destination Vector pB7m34GW,0 in LR buffer and TE buffers Thaw, vortex and add LR Clonase enzyme (LR Clonase Plus enzyme for multi-site Gateway). Incubate >= 1 hr. at 25°. Add Proteinase K for 10 min at 37°. Transform into DH5a E. coli cells (One Shot TOP10 for multi-site Gateway) or any non-F' episome bearing strain. Destination Vector and Entry Clone carry identical resistance markers or Destination vector is not adequately annotated. Check annotation and if necessary use a different vector or construct the Entry Clone from a pDONR with different resistance. Express in appropriate host cells. 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|6771 56|7918 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|0 55|6435 56|6767 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|1 55|5180 56|6429 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|0 55|77 56|276 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|0 55|7948 56|8173 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|27 52|M13F 53|1 55|8231 56|8246 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|1 55|33 56|49 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|4 52|mCITRINE 53|1 55|8280 56|8996 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|PH domain AKT 53|1 55|9063 56|9500 57|1 281|1 282|1 283|1 284|1 50 45 51|21 52|GAGA linker 53|1 55|9033 56|9053 57|1 281|1 282|1 283|1 284|1 50 45 51|52 52|5'UTR 53|1 55|9531 56|10030 57|1 281|1 282|1 283|1 284|1 285|1520..2019 50 45 51|21 52|Exon 1.1 53|1 55|9835 56|9919 57|1 281|1 282|1 283|1 284|1 285|1631..1715 50 45 51|15 52|Intron 1.1 53|1 55|9531 56|9834 57|1 281|1 282|1 283|1 284|1 285|1716..2019 50 45 51|29 52|5' Promoter region 53|1 55|9920 56|11516 57|1 281|1 282|1 283|1 284|1 285|34..1630 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|8262 76|1 77|8262 78|2019 79|335 80|0 84 91|2 92|0 93|2019 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|335 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|mCITRINE+STOP/pDONRP2R-P3 72|0 73|0 74|1 75|744 76|8263 77|9006 78|2624 79|5 80|0 84 91|2 92|0 93|2624 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|PH AKT-GAGAnoSTOP/pDONR221 72|0 73|0 74|1 75|510 76|9007 77|9516 78|2523 79|5 80|0 84 91|2 92|0 93|2523 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|1 61|0 63|0 70|UB10-promoteur/pDONRP4-P1R 72|0 73|0 74|1 75|2010 76|9517 77|11526 78|2628 79|5 80|0 84 91|2 92|0 93|2628 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|2 92|1 93|5 96|0 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 1024|-1 24 209|ctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggccgacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgc 209|gtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggccagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagc 209|atcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgccggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcag 209|ccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctca 209|tcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgcattacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaaca 209|ggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagctctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgcttacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactccagcaggaccatgtgatcgcgcttctcgttggggtctttgctcagcttggactggtagctcaggtagtgg 209|ttgtcgggcagcagcacggggccgtcgccgatgggggtgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatcttgaagttcaccttgatgccgttcttctgcttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccaggatgttgccgtcctccttgaagtcgatgcccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtcttgtagttgccgtcgtccttgaagaagatggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttcatgtggtcggggtagcgggcgaagcacatcaggccgtagccgaaggtggtcacgagggtgggccagggcacgggcagcttgccggtggtgcagatgaacttcagggtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccgtttacgtcgccgtccagctcgaccaggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatagccactttgtacaagaaagctgggtactcgagagaaccagcaccagcaccagcacccttgttcatggtcacgcggtgcttgggcttggccagggacacctccatctcttcagcccctgagttgtcactgggtgagcccgaccggaagtccatctcctcctcctcctgcttcttgaggccgtcagccacagtctggatggcggttgtccactcctcccgctcctcaggagtctccacatggaaggtgcgttcgatgacagtggtccactgcaggcagcggatgatgaaggtgttgggccggggccgctccgtcttcatcagctggcactgcgccacagagaagttgttgaggggagcctcacgttggtccacatcctgcggccgctccttgtagccaatgaaggtgccatcattcttgaggaggaagtagcgtggccgccaggtcttgatgtactcccctcgtttgtgcagccaaccctccttcacaatagccacgtcgctcatggttaagcctgcttttttgtacaaacttgcctgttaatcagaaaaactcagattaatcgacaaattcgatcgcacaaactagaaactaacaccagatctagatagaaatcacaaatcgaagagtaattattcgacaaaactcaaattatttgaacaaatcggatgatatctatgaaaccctaatcgagaattaagatgatatctaacgatcaaacccagaaaatcgtcttcgatctaagattaacagaatctaaaccaaagaacatatacgaaattgggatcgaacgaaaacaaaatcga 209|agattttgagagaataaggaacacagaaatttaccttgatcacggtagagagaattgagagaaagtttttaagattttgagaaattgaaatctgaattgtgaagaagaagagctctttgggtattgttttatagaagaagaagaagaaaagacgaggacgactaggtcacgagaaagctaaggcggtgaagcaatagctaataataaaatgacacgtgtattgagcgttgtttacacgcaaagttgtttttggctaattgccttatttttaggttgaggaaaagtatttgtgctttgagttgataaacacgactcgtgtgtgccggctgcaaccactttgacgccgtttattactgactcgtcgacaaccacaatttctaacggtcgtcataagatccagccgttgagatttaacgatcgttacgatttatatttttttagcattatcgttttattttttaaatatacggtggagctgaaaattggcaataattgaaccgtgggtcccactgcattgaagcgtatttcgtattttctagaattcttcgtgctttatttcttttcctttttgtttttttttgccatttatctaatgcaagtgggcttataaaatcagtgaatttcttggaaaagtaacttctttatcgtataacatattgtgaaattatccatttcttttaattttttagtgttattggatatttttgtatgattattgatttgcataggataatgacttttgtatcaagttggtgaacaagtctcgttaaaaaaggcaagtggtttggtgactcgatttattcttgttatttaattcatatatcaatggatcttatttggggcctggtccatatttaacactcgtgttcagtccaatgaccaataatattttttcattaataacaatgtaacaagaatgatacacaaaacattctttgaataagttcgctatgaagaagggaacttatccggtcctagatcatcagttcatacaaacctccatagagttcaacatcttaaacaagaatatcctgatctgaagaatgtggaggctttagtcccttggatacttgggaggctgtggaagaacagaaacgagctggtgcttaaagggagggaatttggaaccaatgaggtattagtaaggacacaagaagatgcagatgagtggattagaaggaaagaggctcagaatgtaaggaaagcaccaaccacaacaccgtcgggacagacgagaaacatgggaagctccaccacaatcttgggttaagtgcaactttgatggggcatggccaacagaaggattaaaatgtggcttagggtgggtgcttcgcgatcatacagggaaggtgttatggttaggtgcaagagctgtggtaaaagtaagaagcgtgctggaa 209|gtagaagtggaggctcttagatgggctgtgctgtcattatcccgattcaattataggaagatcatttttgaggtggattctcagcaacttgtgtctttggttacatgaaagttatgcttgtcaagtctcaatccaattatccaagacataaagtatctacttagcaagtttgaggattttatgcttgtgcatacaagccgagaaggaaatggagtggcagatagaatagctaaggaatctctttcttttgagaattatgatctaaagttgtattctattgtaccaatttgggttaaaagctctgttgagctagactagcaactttt 210 212 25|pB7m34GW,0 278|GenBank 27|0 222|3 33|9945 236|342098876 237|473164835 26|450 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 255 224|Demo User 227|manka@psb.ugent.be 50 256 224|Mansour Karimi 225|+3193313850 227|mansour.karimi@psb.ugent.be 50 1001|0 1006|19-SEP-2007 45 51|27 52|M13 R 53|1 55|2051 56|2067 57|0 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|27 52|M13F 53|1 55|304 56|319 57|0 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|86 52|attR4 53|1 55|1907 56|2031 57|0 281|1 282|1 283|1 284|1 285|complement(1907..2031) 50 45 51|21 52|ccdB 53|1 55|453 56|1906 57|0 281|1 282|1 283|1 284|1 285|complement(453..1906) 50 45 51|86 52|attR3 53|1 55|328 56|452 57|0 281|1 282|1 283|1 284|1 285|complement(328..452) 50 45 51|43 52|T35S 53|0 55|21 56|246 57|0 281|1 282|1 283|1 284|1 285|21..246 50 45 51|87 52|RB 53|0 55|2095 56|2294 57|0 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|22 52|Sm/SpR 53|1 55|7198 56|8447 57|0 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|LB 53|0 55|8453 56|8785 57|0 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Bar 53|1 54|Pnos-Bar-Tnos, Plant selectable marker 55|8789 56|9936 57|0 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pKm34GW7,1 72|0 73|0 74|1 75|2092 76|1 77|2092 78|1 79|2092 80|0 84 91|1 92|0 93|0 94|SacI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|1 92|1 93|0 94|ApaI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|pB+,1 72|0 73|0 74|1 75|7853 76|2093 77|9945 78|1152 79|1124 80|0 84 91|1 92|0 93|0 94|ApaI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|1 92|1 93|0 94|SacI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 250 248|http://www.informaxinc.com/ 50 1024|-1 24 209|ctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgaacgagaaacgtaaaatgatataaatatcaatatattaaattagattttgcataaaaaacagactacataatactgtaaaacacaacatatccagtcactatggcggccgcattaggcacccccaggctttacactttatgcttccggctcgtataatgtgtggattttgagttaggatccgtcgagattttcaggagctaaggaagctaaaatggagaaaaaaatcactggatataccaccgttgatatatcccaatggcatcgtaaagaacattttgaggcatttcagtcagttgctcaatgtacctataaccagaccgttcagctggatattacggcctttttaaagaccgtaaagaaaaataagcacaagttttatccggcctttattcacattcttgcccgcctgatgaatgctcatccggaattccgtatggcaatgaaagacggtgagctggtgatatgggatagtgttcacccttgttacaccgttttccatgagcaaactgaaacgttttcatcgctctggagtgaataccacgacgatttccggcagtttctacacatatattcgcaagatgtggcgtgttacggtgaaaacctggcctatttccctaaagggtttattgagaatatgtttttcgtctcagccaatccctgggtgagtttcaccagttttgatttaaacgtggccaatatggacaacttcttcgcccccgttttcaccatgggcaaatattatacgcaaggcgacaaggtgctgatgccgctggcgattcaggttcatcatgccgtttgtgatggcttccatgtcggcagaatgcttaatgaattacaacagtactgcgatgagtggcagggcggggcgtaatctagaggatccggcttactaaaagccagataacagtatgcgtatttgcgcgctgatttttgcggtataagaatatatactgatatgtatacccgaagtatgtcaaaaagaggtatgctatgaagcagcgtattacagtgacagttgacagcgacagctatcagttgctcaaggcatata 209|tgatgtcaatatctccggttcggtaagcacaaccatgcagaatgaagcccgtcgtctgcgtgccgaacgctggaaagcggaaaatcaggaagggatggctgaggtcgcccggtttattgaaatgaacggctcttttgccgacgagaacaggggctggtgaaatgcagtttaaggtttacacctataaaagagagagccgttatcgtctgtttgtggatgtacagagtgatattattgacacgcccgggcgacggatggtgatccccctggccagtgcacgtctgctgtcagataaagtcccccgtgaactttacccggtggtgcatatcggggatgaaagctggcgcatgatgaccaccgatatggccagtgtgccggtctccgttatcggggaagaagtggctgatctcagccaccgcgaaaatgacatcaaaaacgccattaacctgatgttctggggaatataaatgtcaggctcccttatacacagccagtctgcaggtcgaccatagtgactggatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaacttttctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggcc 209|gacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgcgtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggc 209|cagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagcatcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgc 209|cggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcagccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcg 209|ctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctcatcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgca 209|ttacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacaggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctaggggtcatcagatttcggtgacgggcaggaccggacggggcggcaccggcaggctgaagtccagctgccagaaacccacgtcatgccagttcccgtgcttgaagccggccgcccgcagcatgccgcggggggcatatccgagcgcctcgtgcatgcgcacgctcgggtcgttgggcagcccgatgacagcgaccacgctcttgaagccctgtgcctccagggacttcagcaggtgggtgtagagcgtggagcccagtcccgtccgctggtggcggggggagacgtacacggtcgactcggccgtccagtcgtaggcgttgcgtgccttccagggacccgcgtaggcgatgccggcgacctcgccgtccacctcggcgacgagccagggatagcgctcccgcagacggacgaggtcgtccgtccactcctgcggttcctgcggctcggtacggaagttgaccgtgcttgtctcgatgtagtggttgacgatggtgcagaccgccggcatgtccgcctcggtggcacggcggatgtcggccgggcgtcgttctgggctcatggtagatcccctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgctta 209|gctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagct 210 212 25|pH7m34GW,0 278|GenBank 27|0 222|3 33|10585 236|342095904 237|473164802 26|449 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 255 224|Demo User 227|manka@psb.ugent.be 50 256 224|Mansour Karimi 225|+3193313850 227|mansour.karimi@psb.ugent.be 50 1001|0 1006|19-SEP-2007 45 51|27 52|M13 R 53|1 55|2051 56|2067 57|0 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|27 52|M13F 53|1 55|304 56|319 57|0 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|86 52|attR4 53|1 55|1907 56|2031 57|0 281|1 282|1 283|1 284|1 285|complement(1907..2031) 50 45 51|21 52|ccdB 53|1 55|453 56|1906 57|0 281|1 282|1 283|1 284|1 285|complement(453..1906) 50 45 51|86 52|attR3 53|1 55|328 56|452 57|0 281|1 282|1 283|1 284|1 285|complement(328..452) 50 45 51|43 52|T35S 53|0 55|21 56|246 57|0 281|1 282|1 283|1 284|1 285|21..246 50 45 51|87 52|RB 53|0 55|2095 56|2294 57|0 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|22 52|Sm/SpR 53|1 55|7198 56|8447 57|0 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|LB 53|0 55|8453 56|8785 57|0 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Hyg 53|1 55|8789 56|10575 57|0 281|1 282|1 283|1 284|1 285|complement(8789..10575) 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pKm34GW7,1 72|0 73|0 74|1 75|2092 76|1 77|2092 78|1 79|2092 80|0 84 91|1 92|0 93|0 94|SacI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|1 92|1 93|0 94|ApaI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|pH+,1 72|0 73|0 74|1 75|8493 76|2093 77|10585 78|11 79|8503 80|0 84 91|1 92|0 93|0 94|ApaI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|1 92|1 93|0 94|SacI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 250 248|http://www.informaxinc.com/ 50 1024|-1 24 209|ctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgaacgagaaacgtaaaatgatataaatatcaatatattaaattagattttgcataaaaaacagactacataatactgtaaaacacaacatatccagtcactatggcggccgcattaggcacccccaggctttacactttatgcttccggctcgtataatgtgtggattttgagttaggatccgtcgagattttcaggagctaaggaagctaaaatggagaaaaaaatcactggatataccaccgttgatatatcccaatggcatcgtaaagaacattttgaggcatttcagtcagttgctcaatgtacctataaccagaccgttcagctggatattacggcctttttaaagaccgtaaagaaaaataagcacaagttttatccggcctttattcacattcttgcccgcctgatgaatgctcatccggaattccgtatggcaatgaaagacggtgagctggtgatatgggatagtgttcacccttgttacaccgttttccatgagcaaactgaaacgttttcatcgctctggagtgaataccacgacgatttccggcagtttctacacatatattcgcaagatgtggcgtgttacggtgaaaacctggcctatttccctaaagggtttattgagaatatgtttttcgtctcagccaatccctgggtgagtttcaccagttttgatttaaacgtggccaatatggacaacttcttcgcccccgttttcaccatgggcaaatattatacgcaaggcgacaaggtgctgatgccgctggcgattcaggttcatcatgccgtttgtgatggcttccatgtcggcagaatgcttaatgaattacaacagtactgcgatgagtggcagggcggggcgtaatctagaggatccggcttactaaaagccagataacagtatgcgtatttgcgcgctgatttttgcggtataagaatatatactgatatgtatacccgaagtatgtcaaaaagaggtatgctatgaagcagcgtattacagtgacagttgacagcgacagctatcagttgctcaaggcatata 209|tgatgtcaatatctccggttcggtaagcacaaccatgcagaatgaagcccgtcgtctgcgtgccgaacgctggaaagcggaaaatcaggaagggatggctgaggtcgcccggtttattgaaatgaacggctcttttgccgacgagaacaggggctggtgaaatgcagtttaaggtttacacctataaaagagagagccgttatcgtctgtttgtggatgtacagagtgatattattgacacgcccgggcgacggatggtgatccccctggccagtgcacgtctgctgtcagataaagtcccccgtgaactttacccggtggtgcatatcggggatgaaagctggcgcatgatgaccaccgatatggccagtgtgccggtctccgttatcggggaagaagtggctgatctcagccaccgcgaaaatgacatcaaaaacgccattaacctgatgttctggggaatataaatgtcaggctcccttatacacagccagtctgcaggtcgaccatagtgactggatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaacttttctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggcc 209|gacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgcgtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggc 209|cagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagcatcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgc 209|cggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcagccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcg 209|ctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctcatcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgca 209|ttacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacaggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagggaattaattcctgaatcactgcgaccggccctcccgcgacccagccgagcgagcttagcgaactgtggacgagaactgtgccaccaagcgtaaggccgttctctcgcatttgccttgctaggctcgcgcgagttgctggctgaggcgttctcgaaatcagctcttgttcggtcggcatctactctattcctttgccctcggacgagtgctggggcgtcggtttccactatcggcgagtacttctacacagccatcggtccagacggccgcgcttctgcgggcgatttgtgtacgcccgacagtcccggctccggatcggacgattgcgtcgcatcgaccctgcgcccaagctgcatcatcgaaattgccgtcaaccaagctctgatagagttggtcaagaccaatgcggagcatatacgcccggagccttggcgatcctgcaagctccggatgcctccgctcgaagtagcgcgtctgctgctccatacaagccaaccacggcctccagaagaagatgttggcgacctcgtattgggaatccccgaacatcgcctcgctccagtcaatgaccgctgttatgcggccattgtccgtcaggacattgttggagccgaatccgcgtgcacgagtgccggacttcggggcagtcctcgcccaaagcatcagctcatcgagagcctgcgcgacggacgcactgacggtgtcgtccatcacagtttgccagtg 209|atacacatggggatcagcaatcgcgcatatgaaatcacgccatgtagtgtattgaccgattccttgcggtccgaatgggccgaacccgctcgtctggctaagatcggccgcagcgatagcatccatagcctccgcgaccggctgaagaacagcgggcagttcggtttcaggcaggtcttgcaacgtgacaccctgtgcacggcgggagatgcaataggtcaggctctcgctgaactccccaatgtcaagcacttccggaatcgggagcgcggccgatgcaaagtgccgataaacataacgatctttgtagaaaccatcggcgcagctatttacccgcaggacatatccacgccctcctacatcgaagctgaaagcacgagattcttcgccctccgagagctgcatcaggtcggagacgctgtcgaacttttcgatcagaaacttctcgacagacgtcgcggtgagttcaggctttttcataggggggattcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctcagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcggggtcataacgtgactcccttaattctcatgtatgataattcgagct 210 212 25|pK7m34GW,0 278|GenBank 27|0 222|3 33|10198 236|342098615 237|473164771 26|448 28|0 219|0 220|1 221|1 29|0 30|1 217|0 31|1 32|1 255 224|Demo User 227|manka@psb.ugent.be 50 256 224|Mansour Karimi 225|+3193313850 227|mansour.karimi@psb.ugent.be 50 1001|0 1006|19-SEP-2007 45 51|27 52|M13 R 53|1 55|2051 56|2067 57|0 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 45 51|27 52|M13F 53|1 55|304 56|319 57|0 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|86 52|attR4 53|1 55|1907 56|2031 57|0 281|1 282|1 283|1 284|1 285|complement(1907..2031) 50 45 51|21 52|ccdB 53|1 55|453 56|1906 57|0 281|1 282|1 283|1 284|1 285|complement(453..1906) 50 45 51|86 52|attR3 53|1 55|328 56|452 57|0 281|1 282|1 283|1 284|1 285|complement(328..452) 50 45 51|43 52|T35S 53|0 55|21 56|246 57|0 281|1 282|1 283|1 284|1 285|21..246 50 45 51|87 52|RB 53|0 55|2095 56|2294 57|0 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|22 52|Sm/SpR 53|1 55|7198 56|8447 57|0 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|LB 53|0 55|8453 56|8785 57|0 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Kan 53|1 55|8789 56|10185 57|0 281|1 282|1 283|1 284|1 285|complement(8789..10185) 50 205 47 59|3 218|1 60|0 61|0 63|0 70|pKm34GW7,1 72|0 73|0 74|1 75|2092 76|1 77|2092 78|1 79|2092 80|0 84 91|1 92|0 93|0 94|SacI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|1 92|1 93|0 94|ApaI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 47 59|3 218|1 60|0 61|0 63|0 70|pK+,1 72|0 73|0 74|1 75|8106 76|2093 77|10198 78|1425 79|1397 80|0 84 91|1 92|0 93|0 94|ApaI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 85 91|1 92|1 93|0 94|SacI 96|1 97|0 98|0 99|0 100|0 101|0 102|0 103|0 104|0 105|0 50 50 37|0 38|0 39|0 40|0 250 248|http://www.informaxinc.com/ 50 1024|-1 24 209|ctcccatatggtcgacctgcaggtcactggattttggttttaggaattagaaattttattgatagaagtattttacaaatacaaatacatactaagggtttcttatatgctcaacacatgagcgaaaccctataagaaccctaattcccttatctgggaactactcacacattattctggagaaaaatagagagagatagatttgtagagagagactggtgatttttgcggactctagcatggccgctcccggccgccatggccgcggctagtcagttagatcgttttcccagtcacgacgttgtaaaacgacggccagtgaattatcaactatgtataataaagttgaacgagaaacgtaaaatgatataaatatcaatatattaaattagattttgcataaaaaacagactacataatactgtaaaacacaacatatccagtcactatggcggccgcattaggcacccccaggctttacactttatgcttccggctcgtataatgtgtggattttgagttaggatccgtcgagattttcaggagctaaggaagctaaaatggagaaaaaaatcactggatataccaccgttgatatatcccaatggcatcgtaaagaacattttgaggcatttcagtcagttgctcaatgtacctataaccagaccgttcagctggatattacggcctttttaaagaccgtaaagaaaaataagcacaagttttatccggcctttattcacattcttgcccgcctgatgaatgctcatccggaattccgtatggcaatgaaagacggtgagctggtgatatgggatagtgttcacccttgttacaccgttttccatgagcaaactgaaacgttttcatcgctctggagtgaataccacgacgatttccggcagtttctacacatatattcgcaagatgtggcgtgttacggtgaaaacctggcctatttccctaaagggtttattgagaatatgtttttcgtctcagccaatccctgggtgagtttcaccagttttgatttaaacgtggccaatatggacaacttcttcgcccccgttttcaccatgggcaaatattatacgcaaggcgacaaggtgctgatgccgctggcgattcaggttcatcatgccgtttgtgatggcttccatgtcggcagaatgcttaatgaattacaacagtactgcgatgagtggcagggcggggcgtaatctagaggatccggcttactaaaagccagataacagtatgcgtatttgcgcgctgatttttgcggtataagaatatatactgatatgtatacccgaagtatgtcaaaaagaggtatgctatgaagcagcgtattacagtgacagttgacagcgacagctatcagttgctcaaggcatata 209|tgatgtcaatatctccggttcggtaagcacaaccatgcagaatgaagcccgtcgtctgcgtgccgaacgctggaaagcggaaaatcaggaagggatggctgaggtcgcccggtttattgaaatgaacggctcttttgccgacgagaacaggggctggtgaaatgcagtttaaggtttacacctataaaagagagagccgttatcgtctgtttgtggatgtacagagtgatattattgacacgcccgggcgacggatggtgatccccctggccagtgcacgtctgctgtcagataaagtcccccgtgaactttacccggtggtgcatatcggggatgaaagctggcgcatgatgaccaccgatatggccagtgtgccggtctccgttatcggggaagaagtggctgatctcagccaccgcgaaaatgacatcaaaaacgccattaacctgatgttctggggaatataaatgtcaggctcccttatacacagccagtctgcaggtcgaccatagtgactggatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaacttttctatacaaagttgatagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcgggcccaagcttagcttgagcttggatcagattgtcgtttcccgccttcagtttaaactatcagtgtttgacaggatatattggcgggtaaacctaagagaaaagagcgtttattagaataacggatatttaaaagggcgtgaaaaggtttatccgttcgtccatttgtatgtgcatgccaaccacagggttcccctcgggatcaaagtactttgatccaacccctccgctgctatagtgcagtcggcttctgacgttcagtgcagccgtcttctgaaaacgacatgtcgcacaagtcctaagttacgcgacaggctgccgccctgcccttttcctggcgttttcttgtcgcgtgttttagtcgcataaagtagaatacttgcgactagaaccggagacattacgccatgaacaagagcgccgccgctggcctgctgggctatgcccgcgtcagcaccgacgaccaggacttgaccaaccaacgggccgaactgcacgcggccggctgcaccaagctgttttccgagaagatcaccggcaccaggcgcgaccgcccggagctggccaggatgcttgaccacctacgccctggcgacgttgtgacagtgaccaggctagaccgcctggcccgcagcacccgcgacctactggacattgccgagcgcatccaggaggccggcgcgggcctgcgtagcctggcagagccgtgggcc 209|gacaccaccacgccggccggccgcatggtgttgaccgtgttcgccggcattgccgagttcgagcgttccctaatcatcgaccgcacccggagcgggcgcgaggccgccaaggcccgaggcgtgaagtttggcccccgccctaccctcaccccggcacagatcgcgcacgcccgcgagctgatcgaccaggaaggccgcaccgtgaaagaggcggctgcactgcttggcgtgcatcgctcgaccctgtaccgcgcacttgagcgcagcgaggaagtgacgcccaccgaggccaggcggcgcggtgccttccgtgaggacgcattgaccgaggccgacgccctggcggccgccgagaatgaacgccaagaggaacaagcatgaaaccgcaccaggacggccaggacgaaccgtttttcattaccgaagagatcgaggcggagatgatcgcggccgggtacgtgttcgagccgcccgcgcacgtctcaaccgtgcggctgcatgaaatcctggccggtttgtctgatgccaagctggcggcctggccggccagcttggccgctgaagaaaccgagcgccgccgtctaaaaaggtgatgtgtatttgagtaaaacagcttgcgtcatgcggtcgctgcgtatatgatgcgatgagtaaataaacaaatacgcaaggggaacgcatgaaggttatcgctgtacttaaccagaaaggcgggtcaggcaagacgaccatcgcaacccatctagcccgcgccctgcaactcgccggggccgatgttctgttagtcgattccgatccccagggcagtgcccgcgattgggcggccgtgcgggaagatcaaccgctaaccgttgtcggcatcgaccgcccgacgattgaccgcgacgtgaaggccatcggccggcgcgacttcgtagtgatcgacggagcgccccaggcggcggacttggctgtgtccgcgatcaaggcagccgacttcgtgctgattccggtgcagccaagcccttacgacatatgggccaccgccgacctggtggagctggttaagcagcgcattgaggtcacggatggaaggctacaagcggcctttgtcgtgtcgcgggcgatcaaaggcacgcgcatcggcggtgaggttgccgaggcgctggccgggtacgagctgcccattcttgagtcccgtatcacgcagcgcgtgagctacccaggcactgccgccgccggcacaaccgttcttgaatcagaacccgagggcgacgctgcccgcgaggtccaggcgctggccgctgaaattaaatcaaaactcatttgagttaatgaggtaaagagaaaatgagcaaaagcacaaacacgctaagtgccggccgtccgagcgcacgcagcagcaaggctgcaacgttggc 209|cagcctggcagacacgccagccatgaagcgggtcaactttcagttgccggcggaggatcacaccaagctgaagatgtacgcggtacgccaaggcaagaccattaccgagctgctatctgaatacatcgcgcagctaccagagtaaatgagcaaatgaataaatgagtagatgaattttagcggctaaaggaggcggcatggaaaatcaagaacaaccaggcaccgacgccgtggaatgccccatgtgtggaggaacgggcggttggccaggcgtaagcggctgggttgtctgccggccctgcaatggcactggaacccccaagcccgaggaatcggcgtgacggtcgcaaaccatccggcccggtacaaatcggcgcggcgctgggtgatgacctggtggagaagttgaaggccgcgcaggccgcccagcggcaacgcatcgaggcagaagcacgccccggtgaatcgtggcaagcggccgctgatcgaatccgcaaagaatcccggcaaccgccggcagccggtgcgccgtcgattaggaagccgcccaagggcgacgagcaaccagattttttcgttccgatgctctatgacgtgggcacccgcgatagtcgcagcatcatggacgtggccgttttccgtctgtcgaagcgtgaccgacgagctggcgaggtgatccgctacgagcttccagacgggcacgtagaggtttccgcagggccggccggcatggccagtgtgtgggattacgacctggtactgatggcggtttcccatctaaccgaatccatgaaccgataccgggaagggaagggagacaagcccggccgcgtgttccgtccacacgttgcggacgtactcaagttctgccggcgagccgatggcggaaagcagaaagacgacctggtagaaacctgcattcggttaaacaccacgcacgttgccatgcagcgtacgaagaaggccaagaacggccgcctggtgacggtatccgagggtgaagccttgattagccgctacaagatcgtaaagagcgaaaccgggcggccggagtacatcgagatcgagctagctgattggatgtaccgcgagatcacagaaggcaagaacccggacgtgctgacggttcaccccgattactttttgatcgatcccggcatcggccgttttctctaccgcctggcacgccgcgccgcaggcaaggcagaagccagatggttgttcaagacgatctacgaacgcagtggcagcgccggagagttcaagaagttctgtttcaccgtgcgcaagctgatcgggtcaaatgacctgccggagtacgatttgaaggaggaggcggggcaggctggcccgatcctagtcatgcgctaccgcaacctgatcgagggcgaagcatccgc 209|cggttcctaatgtacggagcagatgctagggcaaattgccctagcaggggaaaaaggtcgaaaaggtctctttcctgtggatagcacgtacattgggaacccaaagccgtacattgggaaccggaacccgtacattgggaacccaaagccgtacattgggaaccggtcacacatgtaagtgactgatataaaagagaaaaaaggcgatttttccgcctaaaactctttaaaacttattaaaactcttaaaacccgcctggcctgtgcataactgtctggccagcgcacagccgaagagctgcaaaaagcgcctacccttcggtcgctgcgctccctacgccccgccgcttcgcgtcggcctatcgcggccgctggccgctcaaaaatggctggcctacggccaggcaatctaccagggcgcggacaagccgcgccgtcgccactcgaccgccggcgcccacatcaaggcaccctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcagccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcg 209|ctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgcatgatatatctcccaatttgtgtagggcttattatgcacgcttaaaaataataaaagcagacttgacctgatagtttggctgtgagcaattatgtgcttagtgcatctaatcgcttgagttaacgccggcgaagcggcgtcggcttgaacgaatttctagctagacattatttgccgactaccttggtgatctcgcctttcacgtagtggacaaattcttccaactgatctgcgcgcgaggccaagcgatcttcttcttgtccaagataagcctgtctagcttcaagtatgacgggctgatactgggccggcaggcgctccattgcccagtcggcagcgacatccttcggcgcgattttgccggttactgcgctgtaccaaatgcgggacaacgtaagcactacatttcgctcatcgccagcccagtcgggcggcgagttccatagcgttaaggtttcatttagcgcctcaaatagatcctgttcaggaaccggatcaaagagttcctccgccgctggacctaccaaggcaacgctatgttctcttgcttttgtcagcaagatagccagatcaatgtcgatcgtggctggctcgaagatacctgcaagaatgtcattgcgctgccattctccaaattgcagttcgcgcttagctggataacgccacggaatgatgtcgtcgtgcacaacaatggtgacttctacagcgcggagaatctcgctctctccaggggaagccgaagtttccaaaaggtcgttgatcaaagctcgccgcgttgtttcatcaagccttacggtcaccgtaaccagcaaatcaatatcactgtgtggcttcaggccgccatccactgcggagccgtacaaatgtacggccagcaacgtcggttcgagatggcgctcgatgacgccaactacctctgatagttgagtcgatacttcggcgatcaccgcttcccccatgatgtttaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtaccccaaaaaaacatgtcataacaagaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgacggcagttacgctacttgca 209|ttacagcttacgaaccgaacgaggcttatgtccactgggttcgtgcccgaattgatcacaggcagcaacgctctgtcatcgttacaatcaacatgctaccctccgcgagatcatccgtgtttcaaacccggcagcttagttgccgttcttccgaatagcatcggtaacatgagcaaagtctgccgccttacaacggctctcccgctgacgccgtcccggactgatgggctgcctgtatcgagtggtgattttgtgccgagctgccggtcggggagctgttggctggctggtggcaggatatattgtggtgtaaacaaattgacgcttagacaacttaataacacattgcggacgtttttaatgtactgaattaacgccgaattgaattatcagcttgcatgccggtcgatctagtaacatagatgacaccgcgcgcgataatttatcctagtttgcgcgctatattttgttttctatcgcgtattaaatgtataattgcgggactctaatcataaaaacccatctcataaataacgtcatgcattacatgttaattattacatgcttaacgtaattcaacagaaattatatgataatcatcgcaagaccggcaacaggattcaatcttaagaaactttattgccaaatgtttgaacgatctgcttgactctagctagagtccgaaccccagagtcccgctcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcgaatcgggagcggcgataccgtaaagcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagccaacgctatgtcctgatagcggtccgccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatattcggcaagcaggcatcgccgtgggtcacgacgagatcctcgccgtcgggcatccgcgccttgagcctggcgaacagttcggctggcgcgagcccctgatgctcttcgtccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcgatgtttcgcttggtggtcgaatgggcaggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctcggcaggagcaaggtgagatgacaggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaacgtcgagcacagctgcgcaaggaacgcccgtcgtggccagccacgatagccgcgctgcctcgtcttggagttcattcagggcaccggacaggtcggtcttgacaaaaagaaccgggcgcccctgcgctgacagccggaacacggcggcatcag 209|agcagccgattgtctgttgtgcccagtcatagccgaatagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgcctcgatcgagttgagagtgaatatgagactctaattggataccgaggggaatttatggaacgtcagtggagcatttttgacaagaaatatttgctagctgatagtgaccttaggcgacttttgaacgcgcaataatggtttctgacgtatgtgcttagctcattaaactccagaaacccgcggctgagtggctccttcaacgttgcggttctgtcagttccaaacgtaaaacggcttgtcccgcgtcatcggcgggggtcataacgtgactcccttaattctcatgtatgataattcgagct 210 207