@551656AS34@ Group 1 267|RDP-Z400-SI 212 25|1110943_IRES_NLS_2xYPet_pMK-RQ 278|GenBank 27|0 222|3 33|4034 236|586960999 237|586960999 26|2086 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|RDP-Z400-SI 50 256 224|UNKNOWN 50 1001|0 1006|19-MAY-2008 45 51|33 52|Col E1 origin 53|1 55|2294 56|2961 57|0 281|1 282|1 283|1 284|1 285|complement(2294..2961) 50 45 51|4 52|KanR 53|1 55|3109 56|3906 57|0 281|1 282|1 283|1 284|1 285|complement(3109..3906) 50 45 51|4 52|no_name 53|0 55|370 56|2125 57|0 281|1 282|1 283|1 284|1 285|370..2125 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ctaaattgtaagcgttaatattttgttaaaattcgcgttaaatttttgttaaatcagctcattttttaaccaataggccgaaatcggcaaaatcccttataaatcaaaagaatagaccgagatagggttgagtggccgctacagggcgctcccattcgccattcaggctgcgcaactgttgggaagggcgtttcggtgcgggcctcttcgctattacgccagctggcgaaagggggatgtgctgcaaggcgattaagttgggtaacgccagggttttcccagtcacgacgttgtaaaacgacggccagtgagcgcgacgtaatacgactcactatagggcgaattgaaggaaggccgtcaaggccgcatgagctcggggacagctttcttgtacaaagtggccgaattcgtcgattcggttgcagcatttaaagcggttgacaactttaaaagaaggaaaaagaaggttgaagaaaagggtgtagtaagtaagtataagtacagaccggagaagtacgccggtcctgattcgtttaatttgaaagaagaaaccatgggcaaaggcgaagagctgttcaccggcgtggtgcccatcctggtggagctggacggcgacgtgaacggccacaagttcagcgtgagcggcgagggcgagggcgacgccacctacggcaagctgaccctgaagctgctgtgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgggctacggcgtgcagtgcttcgcccggtaccccgaccacatgaagcagcacgacttcttcaagagcgccatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacaacagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccggcacaacatcgaggacggcggcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagagcgccctgttcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcctgaccgccgccggcatcaccgagggcatgaacgagctgtataatctactagaccctttgcctgctgccgctgccgctgtgagcaaaggcgaagagctgttcaccggcgtggtgcccatcctggtggagctggacggcgacgtgaacggccacaagttcagcgtgagcggcgagggcg 209|agggcgacgccacctacggcaagctgaccctgaagctgctgtgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgggctacggcgtgcagtgcttcgcccggtaccccgaccacatgaagcagcacgacttcttcaagagcgccatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacaacagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccggcacaacatcgaggacggcggcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagagcgccctgttcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcctgaccgccgccggcatcaccgagggcatgaacgagctgtataatctactagacgctgctgctaagagacctgctgctactaagaaggctggacaagctaagaagaagaagttggattaataagcaactttattatacaaagttgtccccaagcttctgggcctcatgggccttcctttcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaacatggtcatagctgtttccttgcgtattgggcgctctccgcttcctcgctcactgactcgctgcgctcggtcgttcgggtaaagcctggggtgcctaatgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtattt 209|ggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttattagaaaaattcatccagcagacgataaaacgcaatacgctggctatccggtgccgcaatgccatacagcaccagaaaacgatccgcccattcgccgcccagttcttccgcaatatcacgggtggccagcgcaatatcctgataacgatccgccacgcccagacggccgcaatcaataaagccgctaaaacggccattttccaccataatgttcggcaggcacgcatcaccatgggtcaccaccagatcttcgccatccggcatgctcgctttcagacgcgcaaacagctctgccggtgccaggccctgatgttcttcatccagatcatcctgatccaccaggcccgcttccatacgggtacgcgcacgttcaatacgatgtttcgcctgatgatcaaacggacaggtcgccgggtccagggtatgcagacgacgcatggcatccgccataatgctcactttttctgccggcgccagatggctagacagcagatcctgacccggcacttcgcccagcagcagccaatcacggcccgcttcggtcaccacatccagcaccgccgcacacggaacaccggtggtggccagccagctcagacgcgccgcttcatcctgcagctcgttcagcgcaccgctcagatcggttttcacaaacagcaccggacgaccctgcgcgctcagacgaaacaccgccgcatcagagcagccaatggtctgctgcgcccaatcatagccaaacagacgttccacccacgctgccgggctacccgcatgcaggccatcctgttcaatcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccac 210 212 25|2xCHERRY-4xMyc/pDONRP2R-P3 (p3') 27|0 222|3 33|4352 236|583746335 237|728650182 26|1983 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL3-attR2 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 50 1001|0 45 51|21 52|attL3 53|1 55|4345 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|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|2xmCHERRY-4xMyc 53|0 55|2634 56|4340 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|0 55|4308 56|4337 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|0 55|4269 56|4298 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|0 55|4230 56|4259 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|Myc 53|0 55|4191 56|4220 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|0 55|4167 56|4181 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|0 55|3438 56|3452 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|0 55|2700 56|2714 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|4xMyc 53|0 55|4182 56|4340 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|0 55|3423 56|4159 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|mCHERRY 53|0 55|2685 56|3421 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|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctatgctcgaggaattctgtaccccgggttcgaaatcgataagcttggatcctctagaccctttgcctgctgccgctgccgctgtgagcaagggcgaggaggataacatggccatcatcaaggagttcatgcgcttcaaggtgcacatggagggctccgtgaacggcca 209|cgagttcgagatcgagggcgagggcgagggccgcccctacgagggcacccagaccgccaagctgaaggtgaccaagggtggccccctgcccttcgcctgggacatcctgtcccctcagttcatgtacggctccaaggcctacgtgaagcaccccgccgacatccccgactacttgaagctgtccttccccgagggcttcaagtgggagcgcgtgatgaacttcgaggacggcggcgtggtgaccgtgacccaggactcctccctgcaggacggcgagttcatctacaaggtgaagctgcgcggcaccaacttcccctccgacggccccgtaatgcagaagaagaccatgggctgggaggcctcctccgagcggatgtaccccgaggacggcgccctgaagggcgagatcaagcagaggctgaagctgaaggacggcggccactacgacgctgaggtcaagaccacctacaaggccaagaagcccgtgcagctgcccggcgcctacaacgtcaacatcaagttggacatcacctcccacaacgaggactacaccatcgtggaacagtacgaacgcgccgagggccgccactccaccggcggcatggacgagctgtacaagggactagaccctttgcctgctgccgctgccgctgtgagcaagggcgaggaggataacatggccatcatcaaggagttcatgcgcttcaaggtgcacatggagggctccgtgaacggccacgagttcgagatcgagggcgagggcgagggccgcccctacgagggcacccagaccgccaagctgaaggtgaccaagggtggccccctgcccttcgcctgggacatcctgtcccctcagttcatgtacggctccaaggcctacgtgaagcaccccgccgacatccccgactacttgaagctgtccttccccgagggcttcaagtgggagcgcgtgatgaacttcgaggacggcggcgtggtgaccgtgacccaggactcctccctgcaggacggcgagttcatctacaaggtgaagctgcgcggcaccaacttcccctccgacggccccgtaatgcagaagaagaccatgggctgggaggcctcctccgagcggatgtaccccgaggacggcgccctgaagggcgagatcaagcagaggctgaagctgaaggacggcggccactacgacgctgaggtcaagaccacctacaaggccaagaagcccgtgcagctgcccggcgcctacaacgtcaacatcaagttggacatcacctcccacaacgaggactacaccatcgtggaacagtacgaacgcgccgagggccgccactccaccggcggcatggacgagctgtacaagggactagaagctgctgctgctgctaaagctatggagcaaaagc 209|tcatttctgaagaggacttgaatgaaatggagcaaaagctcatttctgaagaggacttgaatgaaatggagcaaaagctcatttctgaagaggacttgaatgaaatggagcaaaagctcatttctgaagaggacttgaattgagcaacttta 210 212 25|3xYPET-3xHA/pDONRP2R-P3 (p3')swelline 27|0 222|3 33|5036 236|738090536 237|738090536 26|3909 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL3-attR2 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 50 1001|0 45 51|21 52|attL3 53|1 55|5029 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|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|polyA linker 53|0 55|2691 56|2705 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|0 55|3436 56|3449 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|0 55|4179 56|4193 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|polyA linker 53|0 55|4923 56|4937 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|YPET 53|0 55|4173 56|4910 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|YPET 53|0 55|3429 56|4172 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|YPET 53|0 55|2685 56|3428 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|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctcgtctcgaggaattcggtaccccgggttcgaaatcgataagcttggatcctttgcctgctgctgctgccgctgtgagcaaaggcgaagagctgttcaccggcgtggtgcccatcctggtggagctggacggcgacgtgaacggccacaagttcagcgtgagcggcga 209|gggcgagggcgacgccacctacggcaagctgaccctgaagctgctgtgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgggctacggcgtgcagtgcttcgcccggtaccccgaccacatgaagcagcacgacttcttcaagagcgccatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacaacagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccggcacaacatcgaggacggcggcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagagcgccctgttcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcctgaccgccgccggcatcaccgagggcatgaacgagctgtataatctagatcctttgcctgctgctgctgccgctgtgagcaaaggcgaagagctgttcaccggcgtggtgcccatcctggtggagctggacggcgacgtgaacggccacaagttcagcgtgagcggcgagggcgagggcgacgccacctacggcaagctgaccctgaagctgctgtgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgggctacggcgtgcagtgcttcgcccggtaccccgaccacatgaagcagcacgacttcttcaagagcgccatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacaacagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccggcacaacatcgaggacggcggcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagagcgccctgttcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcctgaccgccgccggcatcaccgagggcatgaacgagctgtataatctagatcctttgcctgctgctgctgccgctgtgagca 209|aaggcgaagagctgttcaccggcgtggtgcccatcctggtggagctggacggcgacgtgaacggccacaagttcagcgtgagcggcgagggcgagggcgacgccacctacggcaagctgaccctgaagctgctgtgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgggctacggcgtgcagtgcttcgcccggtaccccgaccacatgaagcagcacgacttcttcaagagcgccatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacaacagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccggcacaacatcgaggacggcggcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagagcgccctgttcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcctgaccgccgccggcatcaccgagggcatgaacgagctctataagggtctagacttgcctgctgctgctgctgcttacccatacgatgttcctgactatgcgggctatccctatgacgtcccggactatgcaggattgtatccatatgacgttccagattactaagcaacttta 210 212 25|GVG::tE9::6xUASmini35S::mCITRINE/pDONR221 27|0 222|3 33|5138 236|680374471 237|680374471 26|3142 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL1-attL2 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 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|5131 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|0 281|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|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|109 56|125 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|106 56|125 57|0 281|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|0 281|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|0 281|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|0 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|259 56|1068 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|4 52|GVG 53|0 54|GAL4-VP16-GR : himeric transcription factor; contains Saccharomyces cerevisiae GAL4BD, Herpes simplex VP16AD and Rattus norvegicus GR domain 55|2539 56|3837 57|0 281|1 282|1 283|1 284|1 50 45 51|43 52|Pisum sativum RbcS-E9 polyA signal 53|0 54|terminator of pea Rbsc E9 55|3841 56|4163 57|0 281|1 282|1 283|1 284|1 50 45 51|29 52|6xUAS mini35Sprom 53|0 55|4223 56|4391 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|UAS 53|0 55|4223 56|4239 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|UAS 53|0 55|4242 56|4258 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|UAS 53|0 55|4262 56|4278 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|UAS 53|0 55|4283 56|4299 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|UAS 53|0 55|4302 56|4318 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|UAS 53|0 55|4321 56|4337 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|GAL4-DB 53|0 54|GAL4 DNA binding domain (AA1-74 of S. cerevisiae) 55|2539 56|2760 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|VP16 53|0 54|Activation domain of VP16 (Herpes simplex virus virion protein 16 C terminal) 55|2767 56|3000 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|GR 53|0 54|Glucocorticoid receptor from Rattus Norvegicus, Ctemrinal half, AA518-794 55|3007 56|3837 57|0 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|0 55|4413 56|5129 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ttgtacaaagttggcattataagaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttgccatccagctgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaataatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaa 209|ctctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccccaaataatgattttattttgactgatagtgacctgttcgttgcaacacattgatgagcaatgcttttttataatgccaactttgtacaaaaaagcaggcttaaccatgaagctactgtcttctatcgaacaagcatgcgatatttgccgacttaaaaagctcaagtgctccaaagaaaaaccgaagtgcgccaagtgtctgaagaacaactgggagtgtcgctactctcccaaaaccaaaaggtctccgctgactagggcacatctgacagaagtggaatcaaggctagaaagactggaacagctatttctactgatttttcctaggtcgagcgcccccccgaccgatgtcagcctgggggacgagc 209|tccacttagacggcgaggacgtggcgatggcgcatgccgacgcgctagacgatttcgatctggacatgttgggggacggggattccccgggtccgggatttaccccccacgactccgccccctacggcgctctggatatggccgacttcgagtttgagcagatgtttaccgatgcccttggaattgacgagtacggtggggatccaattcagcaagccactgcaggagtctcacaagacacttcggaaaatcctaacaaaacaatagttcctgctgcattaccacagctcacccctaccttggtgtcactgctggaggtgattgaacccgaggtgttgtatgcaggatatgatagctctgttccagattcagcatggagaattatgaccacactcaacatgttaggtgggcgtcaagtgattgcagcagtgaaatgggcaaaggcgataccaggcttcagaaacttacacctggatgaccaaatgaccctgctacagtactcatggatgtttctcatggcatttgccctgggttggagatcatacagacaatcaagtggaaacctgctctgctttgctcctgatctgattattaatgagcagagaatgtctctaccctgcatgtatgaccaatgtaaacacatgctgtttgtctcctctgaattacaaagattgcaggtatcctatgaagagtatctctgtatgaaaaccttactgcttctctcctcagttcctaaggaaggtctgaagagccaagagttatttgatgagattcgaatgacttatatcaaagagctaggaaaagccatcgtcaaaagggaagggaactccagtcagaactggcaacggttttaccaactgacaaagcttctggactccatgcatgaggtggttgagaatctccttacctactgcttccagacatttttggataagaccatgagtattgaattcccagagatgttagctgaaatcatcactaatcagataccaaaatattcaaatggaaatatcaaaaagcttctgtttcatcaaaaatgactcgacctaactgagtaagctagcttgttcgagtattatggcattgggaaaactgtttttcttgtaccatttgttgtgcttgtaatttactgtgttttttattcggttttcgctatcgaactgtgaaatggaaatggatggagaagagttaatgaatgatatggtccttttgttcattctcaaattaatattatttgttttttctcttatttgttgtgtgttgaatttgaaattataagagatatgcaaacattttgttttgagtaaaaatgtgtcaaatcgtggcctctaatgaccgaagttaatatgaggagtaaaacactagatccccaaacaagcttgaaactgaaggcgggaaac 209|gacaatctgatctctagcccatcgggtgacagccctccgagcgggtgacagccctccgaagcgggtgacagccctccgacgtcgggtgacagccctccgagcgggtgacagccctccgagcgggtgacagccctccgctgcagcaagacccttcctctatataaggaagttcatttcatttggagaggacagcccaagcttcgactctaaccatggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccttcggctacggcctgatgtgcttcgcccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccaagctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtcagctttc 210 212 25|H2B/pDONR207 27|0 222|3 33|3827 236|483701071 237|728650519 26|752 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL1-attL2 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 50 1001|0 45 51|21 52|attL1 53|0 55|3261 56|3356 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|attL2 53|1 55|3820 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 pDONR207 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 gentamicin plates. Pick colonies and grow in selection media. 50 45 51|27 52|pDONRT207 reverse primer 53|1 55|113 56|136 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1994000" 50 45 51|27 52|pDONRT207 forward primer 53|0 55|3229 56|3253 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1993000" 50 45 51|43 52|rrnB T2 transcription terminator 53|0 55|3002 56|3029 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|3161 56|3204 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|29 52|Pc promoter 53|1 55|1594 56|1621 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2250000" 50 45 51|4 52|Gm(R) 53|1 55|872 56|1405 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1150000" 50 45 51|4 52|H2B 53|0 55|3363 56|3797 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|linker 53|0 55|3798 56|3815 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ttgtacaaagtgggcattataagaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttgccatccagctgcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaggccgggaagccgatctcggcttgaacgaattgttaggtggcggtacttgggtcgatatcaaagtgcatcacttcttcccgtatgcccaactttgtatagagagccactgcgggatcgtcaccgtaatctgcttgcacgtagatcacataagcaccaagcgcgttggcctcatgcttgaggagattgatgagcgcggtggcaatgccctgcctccggtgctcgccggagactgcgagatcatagatatagatctcactacgcggctgctcaaacctgggcagaacgtaagccgcgagagcgccaacaaccgcttcttggtcgaaggcagcaagcgcgatgaatgtcttactacggagcaagttcccgaggtaatcggagtccggctgatgttgggagtaggtggctacgtctccgaactcacgaccgaaaagatcaagagcagcccgcatggatttgacttggtcagggccgagcctacatgtgcgaatgatgcccatacttgagccacctaactttgttttagggcgactgccctgctgcgtaacatcgttgctgctgcgt 209|aacatcgttgctgctccataacatcaaacatcgacccacggcgtaacgcgcttgctgcttggatgcccgaggcatagactgtacaaaaaaacagtcataacaagccatgaaaaccgccactgcgccgttaccaccgctgcgttcggtcaaggttctggaccagttgcgtgagcgcatacgctacttgcattacagtttacgaaccgaacaggcttatgtcaactgggttcgtgccttcatccgtttccacggtgtgcgtcacccggcaaccttgggcagcagcgaagtcgaggcatttctgtcctggctggcgaacgagcgcaaggtttcggtctccacgcatcgtcaggcattggcggccttgctgttcttctacggcaaggtgctgtgcacggatctgccctggcttcaggagatcggaagacctcggccgtcgcggcgcttgccggtggtgctgaccccggatgaagtggttcgcatcctcggttttctggaaggcgagcatcgtttgttcgcccaggactctagctatagttctagtggttggctactaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgg 209|gtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatctcgggccccaaataatgattttattttgactgatagtgacctgttcgttgcaacaaattgatgagcaatgcttttttataatgccaagtttgtacaaaaaagcaggctaccatggcgaaggcagataagaaacctgcggagaagaaaccggcagagaaaactccggcagtcgaaccagcagcagcggcagagaagaaaccaaaagccggaaagaaactccccaaggaaccagccgtcgccggagacaagaagaagaagagatcaaagaagaacgttgagacatacaagatctacatcttcaaggtgttgaagcaagttcatccagacatcggaatctcaagcaaagccatgggaatcatgaacagtttcatcaacgatatctttgagaagcttgctggtgagtcttcgaagcttgcgaggtacaacaagaagccgacgattacttccagggagattcagactgcggttagacttgtgttgcccggtgagttggcgaaacatgctgtgtctgaagggactaaggcggttacgaagtttacgagttctgctggagctggggatccgtacccagctttc 210 212 25|IRES-2xYPET-NLS 278|GenBank 27|0 222|3 33|1744 236|582042534 237|588016525 26|2176 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 50 1001|0 1006|16-FEB-2011 45 51|87 52|IRES 53|0 55|29 56|178 57|0 281|1 282|1 283|1 284|1 285|29..178 50 45 51|4 52|YPet 53|0 55|929 56|1648 57|0 281|1 282|1 283|1 284|1 285|929..1648 50 45 51|4 52|YPet 53|0 55|179 56|901 57|0 281|1 282|1 283|1 284|1 285|179..901 50 45 51|4 52|NLS bipartite Nuceloplasmin 53|0 55|1658 56|1711 57|0 281|1 282|1 283|1 284|1 285|1658..1711 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ggggacagctttcttgtacaaagtggccgaattcgtcgattcggttgcagcatttaaagcggttgacaactttaaaagaaggaaaaagaaggttgaagaaaagggtgtagtaagtaagtataagtacagaccggagaagtacgccggtcctgattcgtttaatttgaaagaagaaaccatgggcaaaggcgaagagctgttcaccggcgtggtgcccatcctggtggagctggacggcgacgtgaacggccacaagttcagcgtgagcggcgagggcgagggcgacgccacctacggcaagctgaccctgaagctgctgtgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgggctacggcgtgcagtgcttcgcccggtaccccgaccacatgaagcagcacgacttcttcaagagcgccatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacaacagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccggcacaacatcgaggacggcggcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagagcgccctgttcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcctgaccgccgccggcatcaccgagggcatgaacgagctgtataatctactagaccctttgcctgctgccgctgccgctgtgagcaaaggcgaagagctgttcaccggcgtggtgcccatcctggtggagctggacggcgacgtgaacggccacaagttcagcgtgagcggcgagggcgagggcgacgccacctacggcaagctgaccctgaagctgctgtgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgggctacggcgtgcagtgcttcgcccggtaccccgaccacatgaagcagcacgacttcttcaagagcgccatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacaacagccacaacgtgtacatcaccgccgacaagcagaaga 209|acggcatcaaggccaacttcaagatccggcacaacatcgaggacggcggcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagagcgccctgttcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcctgaccgccgccggcatcaccgagggcatgaacgagctgtataatctactagacgctgctgctaagagacctgctgctactaagaaggctggacaagctaagaagaagaagttggattaataagcaactttattatacaaagttgtcccc 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|RDP-Z400-SI 50 256 224|RDP-Z400-SI 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|myristoylated_mCITRINE noSTOP/pDONR221 corrected 27|0 222|3 33|3288 236|499463396 237|526490541 26|1125 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|attL1-attL2 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 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|3281 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|0 281|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|0 281|1 282|1 283|1 284|1 286|/invitrogen="2580000" 50 45 51|30 52|T7 promoter 53|1 55|109 56|125 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="2370000" 50 45 51|27 52|T7 primer 53|1 55|106 56|125 57|0 281|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|0 281|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|0 281|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|0 281|1 282|1 283|1 284|1 286|/invitrogen="1980000" 50 45 51|4 52|Kan(R) 53|0 55|259 56|1068 57|0 281|1 282|1 283|1 284|1 286|/invitrogen="1210000" 50 45 51|4 52|mCITRINE 53|0 55|2560 56|3276 57|0 281|1 282|1 283|1 284|1 50 45 51|4 52|Myristoylation site 53|0 55|2539 56|2559 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ttgtacaaagttggcattataagaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttgccatccagctgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaataatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaa 209|ctctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccccaaataatgattttattttgactgatagtgacctgttcgttgcaacacattgatgagcaatgcttttttataatgccaactttgtacaaaaaagcaggcttaaccatgggaatttgtatgtctagaatggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccttcggctacggcctgatgtgcttcgcccgctaccccgaccacatgaagc 209|agcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccaagctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtacccagctttc 210 212 25|NTF/pDONR221 278|GenBank 27|0 222|3 33|3696 236|621429650 237|622381014 26|2511 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 34|. 255 224|Demo User 225|dzde 226|edzed 228|ezdezd 227|edded 229|dzed 230|dzedezd 50 256 224|UNKNOWN 50 1001|0 1006|24-MAR-2012 1007|. 1015|. 45 51|21 52|NTF 53|0 55|20 56|1168 57|0 281|1 282|1 283|1 284|1 285|20..1168 286|/ApEinfo_fwdcolor="cyan"|/ApEinfo_revcolor="green" 50 45 51|86 52|attP1 53|0 54|attP1 55|3615 56|3696 57|0 281|1 282|1 283|1 284|1 285|3615..3696 286|/ApEinfo_fwdcolor="cyan"|/ApEinfo_revcolor="green" 50 45 51|28 52|M13F primer 53|0 54|M13F primer 55|3582 56|3597 57|0 281|1 282|1 283|1 284|1 285|3582..3597 286|/ApEinfo_fwdcolor="pink"|/ApEinfo_revcolor="pink" 50 45 51|86 52|attP2 53|1 54|attP2 55|1180 56|1268 57|0 281|1 282|1 283|1 284|1 285|complement(1180..1268) 286|/ApEinfo_fwdcolor="cyan"|/ApEinfo_revcolor="green" 50 45 51|28 52|M13R primer 53|1 54|M13R primer 55|1310 56|1326 57|0 281|1 282|1 283|1 284|1 285|complement(1310..1326) 286|/ApEinfo_fwdcolor="pink"|/ApEinfo_revcolor="pink" 50 45 51|60 52|Gene_1 53|0 54|Kanamycin 55|1439 56|2248 57|0 281|1 282|0 283|1 284|1 285|1439..2248 286|/ApEinfo_fwdcolor="#ff0000"|/ApEinfo_revcolor="green" 50 45 51|98 52|Source_1 53|0 55|1 56|3696 57|0 281|1 282|0 283|1 284|1 285|1..3696 286|/dnas_title="pDONR221+NTF" 50 205 37|0 38|0 39|0 40|0 250 248|Insert from New DNA 13 to 1191 248|Backbone from 248|/Users/ullaspedmale/Desktop/VectorNTI/Vectors/pDONR221.gbk 2896 to 248|651 248| 248|ApEinfo:methylated:1 50 1024|-1 24 209|gtacaaaaaagcaggctccatgaatcattcagcgaaaaccacacagaaccgtgttttgtcagtgaagatgtggccaccgagtaagagtacccgtctcatgcttgttgagcggatgaccaagaacattaccaccccttccatcttctccaggaagtacggtcttttgtctgttgaagaggctgagcaagacgccaagcgcatcgaagatttggcctttgctactgccaacaaacacttccagaacgagcctgatggtgatggcacttctgctgttcacgtctatgctaaagaatccagcaagctcatgcttgatgtcatcaaacgtggtccacaggaagaatccgaggttgaggcggccgctgtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgacctacggcgtgcagtgcttcagccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagcacccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtccggagctgcggccgctgccatggctggtggacttaacgatatcttcgaagctcagaagattgaatggcatgaggatactggtggatcttgagacccagctttcttgtacaaagttggcattataagaaagcattgcttatcaatttgttgcaacgaacaggtcactatcagtcaaaataaaatcattatttgccatccagctgatatcccctatagtgagtcgtattacatggtcatagctgtttcctggcagctctggcccgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataa 209|aactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaacgggaaacgtcgaggccgcgattaaattccaacatggatgctgatttatatgggtataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgcttgtatgggaagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgttacagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaagcattttatccgtactcctgatgatgcatggttactcaccactgcgatccccggaaaaacagcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggcagtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgcgtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgattttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaacttttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatttttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccgataccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaacggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttgatgctcgatgagtttttctaatcagaattggttaattggttgtaacactggcagagcattacgctgacttgacgggacggcgcaagctcatgaccaaaatcccttaacgtgagttacgcgtcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgcc 209|acgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccccaaataatgattttattttgactgatagtgacctgttcgttgcaacaaattgatgagcaatgcttttttataatgccaacttt 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|SV40 NLSe-mCITRINE 27|1 222|3 33|753 236|473180321 237|554057014 26|1475 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|0 32|1 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 50 1001|0 45 51|4 52|mCITRINE 53|0 55|34 56|750 57|0 281|1 282|1 283|1 284|1 50 45 51|21 52|SV40 NLS Eirini 53|0 55|1 56|33 57|0 281|1 282|1 283|1 284|1 50 45 51|4 52|SV40e-CITRINE 53|0 55|1 56|753 57|0 281|1 282|1 283|1 284|1 50 205 37|0 38|0 39|0 40|0 1024|-1 24 209|ttgcaacctaagaagaagagaaaggttggaggaatggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccttcggctacggcctgatgtgcttcgcccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccaagctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtaa 210 212 25|IRES_NLS_2xYPet/pDONRP2R-P3 (p3') 27|0 222|3 33|4331 236|587469055 237|587469055 26|2125 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 50 1001|0 45 51|21 52|attL3 53|1 55|4324 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=If necessary, linearize attB sequence 1110943_IRES_NLS_2xYPet_pMK-RQ at a unique REN site outside the attB region. Ethanol precipitate and resuspend. Mix linearized attB sequence 1110943_IRES_NLS_2xYPet_pMK-RQ 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 E. coli or any non-F' episome bearing strain. Select Entry Clones on agar plates with antibiotic kanamycin. 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" 50 205 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|3 218|1 60|0 61|0 63|0 70|1110943_IRES_NLS_2xYPet_pMK-RQ 72|0 73|0 74|1 75|1713 76|2619 77|4331 78|389 79|2101 80|0 84 91|2 92|0 93|389 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|2101 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|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggccgaattcgtcgattcggttgcagcatttaaagcggttgacaactttaaaagaaggaaaaagaaggttgaagaaaagggtgtagtaagtaagtataagtacagaccggagaagtacgccggtcctgattcgtttaatttgaaagaagaaaccatgggcaaaggcgaaga 209|gctgttcaccggcgtggtgcccatcctggtggagctggacggcgacgtgaacggccacaagttcagcgtgagcggcgagggcgagggcgacgccacctacggcaagctgaccctgaagctgctgtgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgggctacggcgtgcagtgcttcgcccggtaccccgaccacatgaagcagcacgacttcttcaagagcgccatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacaacagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccggcacaacatcgaggacggcggcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagagcgccctgttcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcctgaccgccgccggcatcaccgagggcatgaacgagctgtataatctactagaccctttgcctgctgccgctgccgctgtgagcaaaggcgaagagctgttcaccggcgtggtgcccatcctggtggagctggacggcgacgtgaacggccacaagttcagcgtgagcggcgagggcgagggcgacgccacctacggcaagctgaccctgaagctgctgtgcaccaccggcaagctgcccgtgccctggcccaccctggtgaccaccctgggctacggcgtgcagtgcttcgcccggtaccccgaccacatgaagcagcacgacttcttcaagagcgccatgcccgagggctacgtgcaggagcggaccatcttcttcaaggacgacggcaactacaagacccgggccgaggtgaagttcgagggcgacaccctggtgaaccggatcgagctgaagggcatcgacttcaaggaggacggcaacatcctgggccacaagctggagtacaactacaacagccacaacgtgtacatcaccgccgacaagcagaagaacggcatcaaggccaacttcaagatccggcacaacatcgaggacggcggcgtgcagctggccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagagcgccctgttcaaggaccccaacgagaagcgggaccacatggtgctgctggagttcc 209|tgaccgccgccggcatcaccgagggcatgaacgagctgtataatctactagacgctgctgctaagagacctgctgctactaagaaggctggacaagctaagaagaagaagttggattaataagcaacttta 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|RDP-Z400-SI 50 256 224|RDP-Z400-SI 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|mCITRINEwSTOP/pDONRP2R-P3 27|0 222|3 33|3362 236|473187444 237|473187444 26|464 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 50 1001|0 45 51|21 52|attL3 53|1 55|3355 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|4 52|mCITRINE 53|0 55|2634 56|3350 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|mCITRINE 72|0 73|0 74|1 75|720 76|2634 77|3353 78|1 79|720 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|720 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|3354 77|3362 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|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctatggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgt 209|gccctggcccaccctcgtgaccaccttcggctacggcctgatgtgcttcgcccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccaagctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtaagcaacttta 210 212 25|SV40 NLSe-mCITRINE/pDONRP2R-P3 (p3') 27|0 222|3 33|3395 236|554058991 237|554058991 26|1478 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|attL3-attR2 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 50 1001|0 45 51|21 52|attL3 53|1 55|3388 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|SV40 NLS Eirini 53|0 55|2634 56|2666 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|mCITRINE 53|0 55|2667 56|3383 57|1 281|1 282|1 283|1 284|1 50 45 51|4 52|SV40e-CITRINE 53|0 55|2634 56|3386 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|SV40 NLSe-mCITRINE 72|0 73|0 74|1 75|753 76|2634 77|3386 78|1 79|753 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|753 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|3387 77|3395 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|taggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagcattgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatacgcgtaccgctagccaggaagagtttgtagaaacgcaaaaaggccatccgtcaggatggccttctgcttagtttgatgcctggcagtttatggcgggcgtcctgcccgccaccctccgggccgttgcttcacaacgttcaaatccgctcccggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtcttccgactgagcctttcgttttatttgatgcctggcagttccctactctcgcgttaacgctagcatggatgttttcccagtcacgacgttgtaaaacgacggccagtcttaagctcgggccctgcagctctagagctcgaattctacaggtcactaataccatctaagtagttgattcatagtgactgcatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttcttgtacaaagtggctttgcaacctaagaagaagagaaaggttggaggaatggtgagcaagggcgaggagctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccct 209|gaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccttcggctacggcctgatgtgcttcgcccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccaagctgagcaaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtaagcaacttta 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|pB7m34GW,0 rc 278|GenBank 27|0 222|3 33|9945 236|655900045 237|655900046 26|2778 28|0 219|0 220|1 221|1 29|0 30|0 217|0 31|1 32|1 34|Complementary copy of pB7m34GW,0 255 224|RDP-Z400-SI 50 256 224|RDP-Z400-SI 50 1001|0 45 51|22 52|Bar 53|0 54|Pnos-Bar-Tnos, Plant selectable marker 55|10 56|1157 57|1 281|1 282|1 283|1 284|1 285|complement(8789..9936) 50 45 51|87 52|LB 53|1 55|1161 56|1493 57|1 281|1 282|1 283|1 284|1 285|8453..8785 50 45 51|22 52|Sm/SpR 53|0 55|1499 56|2748 57|1 281|1 282|1 283|1 284|1 285|complement(7198..8447) 50 45 51|87 52|RB 53|1 55|7652 56|7851 57|1 281|1 282|1 283|1 284|1 285|2095..2294 50 45 51|43 52|T35S 53|1 55|9700 56|9925 57|1 281|1 282|1 283|1 284|1 285|21..246 50 45 51|86 52|attR3 53|0 55|9494 56|9618 57|1 281|1 282|1 283|1 284|1 285|complement(328..452) 50 45 51|21 52|ccdB 53|0 55|8040 56|9493 57|1 281|1 282|1 283|1 284|1 285|complement(453..1906) 50 45 51|86 52|attR4 53|0 55|7915 56|8039 57|1 281|1 282|1 283|1 284|1 285|complement(1907..2031) 50 45 51|27 52|M13F 53|0 55|9627 56|9642 57|1 281|1 282|1 283|1 284|1 285|complement(304..319) 50 45 51|27 52|M13 R 53|0 55|7879 56|7895 57|1 281|1 282|1 283|1 284|1 285|complement(2051..2067) 50 205 47 59|3 218|1 60|1 61|0 63|0 70|pB7m34GW,0 72|0 73|0 74|1 75|9945 76|1 77|9945 78|1 79|9945 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|9945 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|0 250 248|http://www.informaxinc.com/ 50 1024|-1 24 209|agctcgaattatcatacatgagaattaagggagtcacgttatgacccccgccgatgacgcgggacaagccgttttacgtttggaactgacagaaccgcaacgttgaaggagccactgagccgcgggtttctggagtttaatgagctaagcacatacgtcagaaaccattattgcgcgttcaaaagtcgcctaaggtcactatcagctagcaaatatttcttgtcaaaaatgctccactgacgttccataaattcccctcggtatccaattagagtctcatattcactctcaactcgatcgaggggatctaccatgagcccagaacgacgcccggccgacatccgccgtgccaccgaggcggacatgccggcggtctgcaccatcgtcaaccactacatcgagacaagcacggtcaacttccgtaccgagccgcaggaaccgcaggagtggacggacgacctcgtccgtctgcgggagcgctatccctggctcgtcgccgaggtggacggcgaggtcgccggcatcgcctacgcgggtccctggaaggcacgcaacgcctacgactggacggccgagtcgaccgtgtacgtctccccccgccaccagcggacgggactgggctccacgctctacacccacctgctgaagtccctggaggcacagggcttcaagagcgtggtcgctgtcatcgggctgcccaacgacccgagcgtgcgcatgcacgaggcgctcggatatgccccccgcggcatgctgcgggcggccggcttcaagcacgggaactggcatgacgtgggtttctggcagctggacttcagcctgccggtgccgccccgtccggtcctgcccgtcaccgaaatctgatgacccctagagtcaagcagatcgttcaaacatttggcaataaagtttcttaagattgaatcctgttgccggtcttgcgatgattatcatataatttctgttgaattacgttaagcatgtaataattaacatgtaatgcatgacgttatttatgagatgggtttttatgattagagtcccgcaattatacatttaatacgcgatagaaaacaaaatatagcgcgcaaactaggataaattatcgcgcgcggtgtcatctatatgttactagatcgaccggcatgcaagctgataattcaattcggcgttaattcagtacattaaaaacgtccgcaatgtgttattaagttgtctaagcgtcaatttgtttacaccacaatatatcctgccaccagccagccaacagctccccgaccggcagctcggcacaaaatcaccactcgatacaggcagcccatcagtccgggacggcgtcagcgggagagccgttgtaaggcggcagactttgctcatgttaccgatgctattcggaagaa 209|cggcaactaagctgccgggtttgaaacacggatgatctcgcggagggtagcatgttgattgtaacgatgacagagcgttgctgcctgtgatcaattcgggcacgaacccagtggacataagcctcgttcggttcgtaagctgtaatgcaagtagcgtaactgccgtcacgcaactggtccagaaccttgaccgaacgcagcggtggtaacggcgcagtggcggttttcatggcttcttgttatgacatgtttttttggggtacagtctatgcctcgggcatccaagcagcaagcgcgttacgccgtgggtcgatgtttgatgttatggagcagcaacgatgttacgcagcagggcagtcgccctaaaacaaagttaaacatcatgggggaagcggtgatcgccgaagtatcgactcaactatcagaggtagttggcgtcatcgagcgccatctcgaaccgacgttgctggccgtacatttgtacggctccgcagtggatggcggcctgaagccacacagtgatattgatttgctggttacggtgaccgtaaggcttgatgaaacaacgcggcgagctttgatcaacgaccttttggaaacttcggcttcccctggagagagcgagattctccgcgctgtagaagtcaccattgttgtgcacgacgacatcattccgtggcgttatccagctaagcgcgaactgcaatttggagaatggcagcgcaatgacattcttgcaggtatcttcgagccagccacgatcgacattgatctggctatcttgctgacaaaagcaagagaacatagcgttgccttggtaggtccagcggcggaggaactctttgatccggttcctgaacaggatctatttgaggcgctaaatgaaaccttaacgctatggaactcgccgcccgactgggctggcgatgagcgaaatgtagtgcttacgttgtcccgcatttggtacagcgcagtaaccggcaaaatcgcgccgaaggatgtcgctgccgactgggcaatggagcgcctgccggcccagtatcagcccgtcatacttgaagctagacaggcttatcttggacaagaagaagatcgcttggcctcgcgcgcagatcagttggaagaatttgtccactacgtgaaaggcgagatcaccaaggtagtcggcaaataatgtctagctagaaattcgttcaagccgacgccgcttcgccggcgttaactcaagcgattagatgcactaagcacataattgctcacagccaaactatcaggtcaagtctgcttttattatttttaagcgtgcataataagccctacacaaattgggagatatatcatgcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaa 209|aagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagcggaagagcgcctgatgcggtattttctccttacgcatctgtgcggtatttcacaccgcatatggtgcactctcagtacaatctgctctgatgccgcatagttaagccagtatacactccgctatcgctacgtgactgggtcatggctgcgccccgacacccgccaacacccgctgacgcgccctgacgggcttgtctgctcccggcatccgcttacagacaagctgtgaccgtctccgggagctgcatgtgtcagaggttttcaccgtcatcaccgaaacgcgcgaggcagggtgccttgatgtgggcgccggcggtcgagtggcgacggcgcggcttgtccgcgccctggtagattgcctggccgtaggccagccatttttgagcggccagcggccgcgataggccgacgcgaagcggcggggcgtagggagcgcagcgaccgaagggtaggcgctttttgcagctcttcggctgtgcgctggccagacagttatgcacaggccaggcgggttttaagagttttaataagttttaaagagttttaggcggaaaaatcgccttttttctcttttatatcagtcacttacatgtgtgaccggttcccaatgtacggctt 209|tgggttcccaatgtacgggttccggttcccaatgtacggctttgggttcccaatgtacgtgctatccacaggaaagagaccttttcgacctttttcccctgctagggcaatttgccctagcatctgctccgtacattaggaaccggcggatgcttcgccctcgatcaggttgcggtagcgcatgactaggatcgggccagcctgccccgcctcctccttcaaatcgtactccggcaggtcatttgacccgatcagcttgcgcacggtgaaacagaacttcttgaactctccggcgctgccactgcgttcgtagatcgtcttgaacaaccatctggcttctgccttgcctgcggcgcggcgtgccaggcggtagagaaaacggccgatgccgggatcgatcaaaaagtaatcggggtgaaccgtcagcacgtccgggttcttgccttctgtgatctcgcggtacatccaatcagctagctcgatctcgatgtactccggccgcccggtttcgctctttacgatcttgtagcggctaatcaaggcttcaccctcggataccgtcaccaggcggccgttcttggccttcttcgtacgctgcatggcaacgtgcgtggtgtttaaccgaatgcaggtttctaccaggtcgtctttctgctttccgccatcggctcgccggcagaacttgagtacgtccgcaacgtgtggacggaacacgcggccgggcttgtctcccttcccttcccggtatcggttcatggattcggttagatgggaaaccgccatcagtaccaggtcgtaatcccacacactggccatgccggccggccctgcggaaacctctacgtgcccgtctggaagctcgtagcggatcacctcgccagctcgtcggtcacgcttcgacagacggaaaacggccacgtccatgatgctgcgactatcgcgggtgcccacgtcatagagcatcggaacgaaaaaatctggttgctcgtcgcccttgggcggcttcctaatcgacggcgcaccggctgccggcggttgccgggattctttgcggattcgatcagcggccgcttgccacgattcaccggggcgtgcttctgcctcgatgcgttgccgctgggcggcctgcgcggccttcaacttctccaccaggtcatcacccagcgccgcgccgatttgtaccgggccggatggtttgcgaccgtcacgccgattcctcgggcttgggggttccagtgccattgcagggccggcagacaacccagccgcttacgcctggccaaccgcccgttcctccacacatggggcattccacggcgtcggtgcctggttgttcttgattttccatgccgcctcctttagccgctaaaattcatctactcatttattcatttgctcat 209|ttactctggtagctgcgcgatgtattcagatagcagctcggtaatggtcttgccttggcgtaccgcgtacatcttcagcttggtgtgatcctccgccggcaactgaaagttgacccgcttcatggctggcgtgtctgccaggctggccaacgttgcagccttgctgctgcgtgcgctcggacggccggcacttagcgtgtttgtgcttttgctcattttctctttacctcattaactcaaatgagttttgatttaatttcagcggccagcgcctggacctcgcgggcagcgtcgccctcgggttctgattcaagaacggttgtgccggcggcggcagtgcctgggtagctcacgcgctgcgtgatacgggactcaagaatgggcagctcgtacccggccagcgcctcggcaacctcaccgccgatgcgcgtgcctttgatcgcccgcgacacgacaaaggccgcttgtagccttccatccgtgacctcaatgcgctgcttaaccagctccaccaggtcggcggtggcccatatgtcgtaagggcttggctgcaccggaatcagcacgaagtcggctgccttgatcgcggacacagccaagtccgccgcctggggcgctccgtcgatcactacgaagtcgcgccggccgatggccttcacgtcgcggtcaatcgtcgggcggtcgatgccgacaacggttagcggttgatcttcccgcacggccgcccaatcgcgggcactgccctggggatcggaatcgactaacagaacatcggccccggcgagttgcagggcgcgggctagatgggttgcgatggtcgtcttgcctgacccgcctttctggttaagtacagcgataaccttcatgcgttccccttgcgtatttgtttatttactcatcgcatcatatacgcagcgaccgcatgacgcaagctgttttactcaaatacacatcacctttttagacggcggcgctcggtttcttcagcggccaagctggccggccaggccgccagcttggcatcagacaaaccggccaggatttcatgcagccgcacggttgagacgtgcgcgggcggctcgaacacgtacccggccgcgatcatctccgcctcgatctcttcggtaatgaaaaacggttcgtcctggccgtcctggtgcggtttcatgcttgttcctcttggcgttcattctcggcggccgccagggcgtcggcctcggtcaatgcgtcctcacggaaggcaccgcgccgcctggcctcggtgggcgtcacttcctcgctgcgctcaagtgcgcggtacagggtcgagcgatgcacgccaagcagtgcagccgcctctttcacggtgcggccttcctggtcgatcagctcgcgggcgtgcgcgatctgtgccggggtga 209|gggtagggcgggggccaaacttcacgcctcgggccttggcggcctcgcgcccgctccgggtgcggtcgatgattagggaacgctcgaactcggcaatgccggcgaacacggtcaacaccatgcggccggccggcgtggtggtgtcggcccacggctctgccaggctacgcaggcccgcgccggcctcctggatgcgctcggcaatgtccagtaggtcgcgggtgctgcgggccaggcggtctagcctggtcactgtcacaacgtcgccagggcgtaggtggtcaagcatcctggccagctccgggcggtcgcgcctggtgccggtgatcttctcggaaaacagcttggtgcagccggccgcgtgcagttcggcccgttggttggtcaagtcctggtcgtcggtgctgacgcgggcatagcccagcaggccagcggcggcgctcttgttcatggcgtaatgtctccggttctagtcgcaagtattctactttatgcgactaaaacacgcgacaagaaaacgccaggaaaagggcagggcggcagcctgtcgcgtaacttaggacttgtgcgacatgtcgttttcagaagacggctgcactgaacgtcagaagccgactgcactatagcagcggaggggttggatcaaagtactttgatcccgaggggaaccctgtggttggcatgcacatacaaatggacgaacggataaaccttttcacgcccttttaaatatccgttattctaataaacgctcttttctcttaggtttacccgccaatatatcctgtcaaacactgatagtttaaactgaaggcgggaaacgacaatctgatccaagctcaagctaagcttgggcccgagcggataacaatttcacacaggaaacagctatgaccatgattacgccaagctatcaactttgtatagaaaagttgaacgagaaacgtaaaatgatataaatatcaatatattaaattagattttgcataaaaaacagactacataatactgtaaaacacaacatatccagtcactatggtcgacctgcagactggctgtgtataagggagcctgacatttatattccccagaacatcaggttaatggcgtttttgatgtcattttcgcggtggctgagatcagccacttcttccccgataacggagaccggcacactggccatatcggtggtcatcatgcgccagctttcatccccgatatgcaccaccgggtaaagttcacgggggactttatctgacagcagacgtgcactggccagggggatcaccatccgtcgcccgggcgtgtcaataatatcactctgtacatccacaaacagacgataacggctctctcttttataggtgtaaaccttaaactgcatttcaccagcccctgtt 209|ctcgtcggcaaaagagccgttcatttcaataaaccgggcgacctcagccatcccttcctgattttccgctttccagcgttcggcacgcagacgacgggcttcattctgcatggttgtgcttaccgaaccggagatattgacatcatatatgccttgagcaactgatagctgtcgctgtcaactgtcactgtaatacgctgcttcatagcatacctctttttgacatacttcgggtatacatatcagtatatattcttataccgcaaaaatcagcgcgcaaatacgcatactgttatctggcttttagtaagccggatcctctagattacgccccgccctgccactcatcgcagtactgttgtaattcattaagcattctgccgacatggaagccatcacaaacggcatgatgaacctgaatcgccagcggcatcagcaccttgtcgccttgcgtataatatttgcccatggtgaaaacgggggcgaagaagttgtccatattggccacgtttaaatcaaaactggtgaaactcacccagggattggctgagacgaaaaacatattctcaataaaccctttagggaaataggccaggttttcaccgtaacacgccacatcttgcgaatatatgtgtagaaactgccggaaatcgtcgtggtattcactccagagcgatgaaaacgtttcagtttgctcatggaaaacggtgtaacaagggtgaacactatcccatatcaccagctcaccgtctttcattgccatacggaattccggatgagcattcatcaggcgggcaagaatgtgaataaaggccggataaaacttgtgcttatttttctttacggtctttaaaaaggccgtaatatccagctgaacggtctggttataggtacattgagcaactgactgaaatgcctcaaaatgttctttacgatgccattgggatatatcaacggtggtatatccagtgatttttttctccattttagcttccttagctcctgaaaatctcgacggatcctaactcaaaatccacacattatacgagccggaagcataaagtgtaaagcctgggggtgcctaatgcggccgccatagtgactggatatgttgtgttttacagtattatgtagtctgttttttatgcaaaatctaatttaatatattgatatttatatcattttacgtttctcgttcaactttattatacatagttgataattcactggccgtcgttttacaacgtcgtgactgggaaaacgatctaactgactagccgcggccatggcggccgggagcggccatgctagagtccgcaaaaatcaccagtctctctctacaaatctatctctctctatttttctccagaataatgtgtgagtagttcccagataaggga 209|attagggttcttatagggtttcgctcatgtgttgagcatataagaaacccttagtatgtatttgtatttgtaaaatacttctatcaataaaatttctaattcctaaaaccaaaatccagtgacctgcaggtcgaccatatgggag 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 210 207