IBMc394 1 pSEVA321-PBAD(SapI-)-B30-His-T7RNAP(29-1)-14aa-FRB-T ds-DNA pSEVA321-PBAD(SapI-)-B30-His-T7RNAP(29-1)-14aa-FRB-T Expression plasmid for split, evolved T7 RNA polymerase N-lobe fused to FRB Creator: Trevor Y. H. Ho Principal Investigator: Baojun Wang BioSafety Level: Level 1 Backbone: pSEVA321 Origin of replication: RK2 Selection Markers: Chloramphenicol 18-NOV-2020 Baojun Wang Trevor Y. H. Ho annotation11 1 BBa_B0030 #c7b0e3 #c7b0e3 BBa_B0030 range 1 1276 1290 annotation5 1 araC #75c6a9 #75c6a9 araC range 1 78 956 annotation15 1 PBAD(SapI-) #faac61 #faac61 PBAD(SapI-) range 1 984 1267 annotation9 1 RFC10 Prefix (full) #d6b295 #d6b295 RFC10 Prefix (full) range 1 56 77 annotation7 1 RFC10 Suffix #9eafd2 #9eafd2 RFC10 Suffix range 1 2237 2257 annotation6 1 cat #ff9ccd #ff9ccd cat range 1 2557 3216 annotation0 1 lambda t0 terminator #c6c9d1 #c6c9d1 lambda t0 terminator range 1 2312 2406 annotation10 1 pSEVA PS2 Rev #85dae9 #85dae9 pSEVA PS2 Rev range 1 2366 2382 annotation12 1 ECK120033736 #d59687 #d59687 ECK120033736 range 1 2184 2236 annotation4 1 cat promoter #c6c9d1 #c6c9d1 cat promoter range 1 2459 2556 annotation13 1 29-1 PACE T7 RNAP 1-179 variant: L32S, E35G, A83A, K98R, L106L, Q107K, T122S, A144T #faac61 #faac61 29-1 PACE T7 RNAP 1-179 variant: L32S, E35G, A83A, K98R, L106L, Q107K, T122S, A144T range 1 1318 1851 annotation1 1 H6 #b7e6d7 #b7e6d7 H6 range 1 1300 1317 annotation8 1 pSEVA PS1 Fwd #85dae9 #85dae9 pSEVA PS1 Fwd range 1 5723 5740 annotation2 1 FRB E coli optimized #b7e6d7 #b7e6d7 FRB E coli optimized range 1 1894 2172 annotation14 1 trfA CDS (replication protein) #b4abac #b4abac trfA CDS (replication protein) range 1 3534 4682 annotation3 1 Long Linker #c6c9d1 #c6c9d1 Long Linker range 1 1852 1893 IBMc394_seq 1 ttaattaaagcggataacaatttcacacaggaggccgcctaggccgcggccgcgcgaattcgcggccgcttctagagttatgacaacttgacggctacatcattcactttttcttcacaaccggcacggaactcgctcgggctggccccggtgcattttttaaatacccgcgagaaatagagttgatcgtcaaaaccaacattgcgaccgacggtggcgataggcatccgggtggtgctcaaaagcagcttcgcctggctgatacgttggtcctcgcgccagcttaagacgctaatccctaactgctggcggaaaagatgtgacagacgcgacggcgacaagcaaacatgctgtgcgacgctggcgatatcaaaattgctgtctgccaggtgatcgctgatgtactgacaagcctcgcgtacccgattatccatcggtggatggagcgactcgttaatcgcttccatgcgccgcagtaacaattgctcaagcagatttatcgccagcagctccgaatagcgcccttccccttgcccggcgttaatgatttgcccaaacaggtcgctgaaatgcggctggtgcgcttcatccgggcgaaagaaccccgtattggcaaatattgacggccagttaagccattcatgccagtaggcgcgcggacgaaagtaaacccactggtgataccattcgcgagcctccggatgacgaccgtagtgatgaatctctcctggcgggaacagcaaaatatcacccggtcggcaaacaaattctcgtccctgatttttcaccaccccctgaccgcgaatggtgagattgagaatataacctttcattcccagcggtcggtcgataaaaaaatcgagataaccgttggcctcaatcggcgttaaacccgccaccagatgggcattaaacgagtatcccggcagcaggggatcattttgcgcttcagccatacttttcatactcccgccattcagagaagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggcActtctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccattactagagattaaagaggagaaaTACTAGATGcatcatcaccatcaccacaacacgattaacatcgctaagaacgacttctctgacatcgaactggctgctatcccgttcaacactctggctgaccattacggtgagcgtTCAgctcgcGGAcagttggcccttgagcatgagtcttacgagatgggtgaagcacgcttccgcaagatgtttgagcgtcaacttaaagctggtgaggttgcggataacgctgccgccaagcctctcatcactaccctactccctaagatgattGCCcgcatcaacgactggtttgaggaagtgaaagctaagcgcggcAGGcgcccgacagccttccagttcCTAAAAgaaatcaagccggaagccgtagcgtacatcaccattaagaccTCTctggcttgcctaaccagtgctgacaatacaaccgttcaggctgtagcaagcgcaatcggtcggACCattgaggacgaggctcgcttcggtcgtatccgtgaccttgaagctaagcacttcaagaaaaacgttgaggaacaactcaacaagcgcgtagggcacgtctacaagGGTGGCTCTGGCTCGAGCGGTGGCTCTGGCTCTGGCTCGAGCGCCGTGTTATGGCACGAACAGTGGTATGAGGGTTTGGATGACGCATCGCGCCAGTTCTTCGGAGAACATAACACCGAAAAAATGTTTGCGGCACTGGAACCTCTTTATGAGATGCTTAAACGTGGGCCTGAAACGTTGCGTGAGATTTCTTTTCAGAACTCATTTGGGCGTGATTTGAACGATGCCTACGAATGGCTGATGAATTACAAAAAGTCGAAAGATGTATCTAACCTGAATCAAGCATGGGATATTTACTATAATGTCTTTCGTAAAATCGGCtaatactagagaacgcatgagAAAGCCCCCGGAAGATCACCTTCCGGGGGCTTTtttattgcgctactagtagcggccgctgcaggcatgcaagcttgcggccgcgtcgtgactgggaaaaccctggcgactagtcttggactcctgttgatagatccagtaatgacctcagaactccatctggatttgttcagaacgctcggttgccgccgggcgttttttattggtgagaatccaggggtccccaataattacgatttaaattggcgaaaatgagacgttgatcggcacgtaagaggttccaactttcaccataatgaaataagatcactaccgggcgtattttttgagttatcgagattttcaggagctaaggaagctaaaatggagaaaaaaatcactggatataccaccgttgatatatcccaatggcatcgtaaagaacattttgaggcatttcagtcagttgctcaatgtacctataaccagaccgttcagctggatattacggcctttttaaagaccgtaaagaaaaataagcacaagttttatccggcctttattcacattcttgcccgcctgatgaatgctcatccggaatttcgtatggcaatgaaagacggtgagctggtgatatgggatagtgttcacccttgttacaccgttttccatgagcaaactgaaacgttttcatcgctctggagtgaataccacgacgatttccggcagtttctacacatatattcgcaagatgtggcgtgttacggtgaaaacctggcctatttccctaaagggtttattgagaatatgtttttcgtctcagccaatccctgggtgagtttcaccagttttgatttaaacgtggccaatatggacaacttcttcgcccccgttttcaccatgggcaaatattatacgcaaggcgacaaggtgctgatgccgctggcgattcaggttcatcatgccgtttgtgatggcttccatgtcggcagaatgcttaatgaattacaacagtactgcgatgagtggcagggcggggcgtaatttgacttttgtccttttccgctgcataaccctgcttcggggtcattatagcgattttttcggtatatccatcctttttcgcacgatatacaggattttgccaaagggttcgtgtagactttccttggtgtatccaacggcgtcagccgggcaggataggtgaagtaggcccacccgcgagcgggtgttccttcttcactgtcccttattcgcacctggcggtgctcaacgggaatcctgctctgcgaggctggccgtaggccggccgcgatgcaggtggctgctgaacccccagccggaactgaccccacaaggccctagcgtttgcaatgcaccaggtcatcattgacccaggcgtgttccaccaggccgctgcctcgcaactcttcgcaggcttcgccgacctgctcgcgccacttcttcacgcgggtggaatccgatccgcacatgaggcggaaggtttccagcttgagcgggtacggctcccggtgcgagctgaaatagtcgaacatccgtcgggccgtcggcgacagcttgcggtacttctcccatatgaatttcgtgtagtggtcgccagcaaacagcacgacgatttcctcgtcgatcaggacctggcaacgggacgttttcttgccacggtccaggacgcggaagcggtgcagcagcgacaccgattccaggtgcccaacgcggtcggacgtgaagcccatcgccgtcgcctgtaggcgcgacaggcattcctcggccttcgtgtaataccggccattgatcgaccagcccaggtcctggcaaagctcgtagaacgtgaaggtgatcggctcgccgataggggtgcgcttcgcgtactccaacacctgctgccacaccagttcgtcatcgtcggcccgcagctcgacgccggtgtaggtgatcttcacgtccttgttgacgtggaaaatgaccttgttttgcagcgcctcgcgcgggattttcttgttgcgcgtggtgaacagggcagagcgggccgtgtcgtttggcatcgctcgcatcgtgtccggccacggcgcaatatcgaacaaggaaagctgcatttccttgatctgctgcttcgtgtgtttcagcaacgcggcctgcttggcttcgctgacctgttttgccaggtcctcgccggcggtttttcgcttcttggtcgtcatagttcctcgcgtgtcgatggtcatcgacttcgccaaacctgccgcctcctgttcgagacgacgcgaacgctccacggcggccgatggcgcgggcagggcagggggagccagttgcacgctgtcgcgctcgatcttggccgtagcttgctggactatcgagccgacggactggaaggtttcgcggggcgcacgcatgacggtgcggcttgcgatggtttcggcatcctcggcggaaaaccccgcgtcgatcagttcttgcctgtatgccttccggtcaaacgtccgattcattcaccctccttgcgggattgccccggaattaattccccggatcgatccgtcgatcttgatcccctgcgccatcagatccttggcggcaagaaagccatccagtttactttgcagggcttcccaaccttaccagagggcgccccagctggcaattccggttcgcttgctgtccataaaaccgcccagtctagctatcgccatgtaagcccactgcaagctacctgctttctctttgcgcttgcgttttcccttgtccagatagcccagtagctgacattcatccggggtcagcaccgtttctgcggactggctttctacgtggctgccatttttggggtgaggccgttcgcggccgaggggcgcagcccctggggggatgggaggcccgcgttagcgggccgggagggttcgagaagggggggcaccccccttcggcgtgcgcggtcacgcgcacagggcgcagccctggttaaaaacaaggtttataaatattggtttaaaagcaggttaaaagacaggttagcggtggccgaaaaacgggcggaaacccttgcaaatgctggattttctgcctgtggacagcccctcaaatgtcaataggtgcgcccctcatctgtcagcactctgcccctcaagtgtcaaggatcgcgcccctcatctgtcagtagtcgcgcccctcaagtgtcaataccgcagggcacttatccccaggcttgtccacatcatctgtgggaaactcgcgtaaaatcaggcgttttcgccgatttgcgaggctggccagctccacgtcgccggccgaaatcgagcctgcccctcatctgtcaacgccgcgccgggtgagtcggcccctcaagtgtcaacgtccgcccctcatctgtcagtgagggccaagttttccgcgaggtatccacaacgccggcggccctacatggctctgctgtagtgagtgggttgcgctccggcagcggtcctgatcccccgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggcgcgcccagctgtctagggcggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtctttcgactgagcctttcgttttatttgatgcct