IBMc396 1 pSEVA321-PBAD(SapI-)-B30-His-T7RNAP(29-1)-15aa-acVHH-T ds-DNA pSEVA321-PBAD(SapI-)-B30-His-T7RNAP(29-1)-15aa-acVHH-T Expression plasmid for split, evolved T7 RNA polymerase N-lobe fused to acVHH 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 3x(GGGGS) linker #b1ff67 #b1ff67 3x(GGGGS) linker range 1 1852 1896 annotation5 1 PBAD(SapI-) #faac61 #faac61 PBAD(SapI-) range 1 984 1267 annotation6 1 cat #ff9ccd #ff9ccd cat range 1 2638 3297 annotation10 1 RFC10 Suffix #9eafd2 #9eafd2 RFC10 Suffix range 1 2318 2338 annotation2 1 acVHH #d59687 #d59687 acVHH range 1 1897 2253 annotation3 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 annotation7 1 RFC10 Prefix (full) #d6b295 #d6b295 RFC10 Prefix (full) range 1 56 77 annotation13 1 H6 #b7e6d7 #b7e6d7 H6 range 1 1300 1317 annotation0 1 trfA CDS (replication protein) #b4abac #b4abac trfA CDS (replication protein) range 1 3615 4763 annotation9 1 araC #75c6a9 #75c6a9 araC range 1 78 956 annotation1 1 ECK120033736 #d59687 #d59687 ECK120033736 range 1 2265 2317 annotation4 1 pSEVA PS2 Rev #85dae9 #85dae9 pSEVA PS2 Rev range 1 2447 2463 annotation8 1 pSEVA PS1 Fwd #85dae9 #85dae9 pSEVA PS1 Fwd range 1 5804 5821 annotation12 1 BBa_B0030 #c7b0e3 #c7b0e3 BBa_B0030 range 1 1276 1290 IBMc396_seq 1 ttaattaaagcggataacaatttcacacaggaggccgcctaggccgcggccgcgcgaattcgcggccgcttctagagttatgacaacttgacggctacatcattcactttttcttcacaaccggcacggaactcgctcgggctggccccggtgcattttttaaatacccgcgagaaatagagttgatcgtcaaaaccaacattgcgaccgacggtggcgataggcatccgggtggtgctcaaaagcagcttcgcctggctgatacgttggtcctcgcgccagcttaagacgctaatccctaactgctggcggaaaagatgtgacagacgcgacggcgacaagcaaacatgctgtgcgacgctggcgatatcaaaattgctgtctgccaggtgatcgctgatgtactgacaagcctcgcgtacccgattatccatcggtggatggagcgactcgttaatcgcttccatgcgccgcagtaacaattgctcaagcagatttatcgccagcagctccgaatagcgcccttccccttgcccggcgttaatgatttgcccaaacaggtcgctgaaatgcggctggtgcgcttcatccgggcgaaagaaccccgtattggcaaatattgacggccagttaagccattcatgccagtaggcgcgcggacgaaagtaaacccactggtgataccattcgcgagcctccggatgacgaccgtagtgatgaatctctcctggcgggaacagcaaaatatcacccggtcggcaaacaaattctcgtccctgatttttcaccaccccctgaccgcgaatggtgagattgagaatataacctttcattcccagcggtcggtcgataaaaaaatcgagataaccgttggcctcaatcggcgttaaacccgccaccagatgggcattaaacgagtatcccggcagcaggggatcattttgcgcttcagccatacttttcatactcccgccattcagagaagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggcActtctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccattactagagattaaagaggagaaaTACTAGATGcatcatcaccatcaccacaacacgattaacatcgctaagaacgacttctctgacatcgaactggctgctatcccgttcaacactctggctgaccattacggtgagcgtTCAgctcgcGGAcagttggcccttgagcatgagtcttacgagatgggtgaagcacgcttccgcaagatgtttgagcgtcaacttaaagctggtgaggttgcggataacgctgccgccaagcctctcatcactaccctactccctaagatgattGCCcgcatcaacgactggtttgaggaagtgaaagctaagcgcggcAGGcgcccgacagccttccagttcCTAAAAgaaatcaagccggaagccgtagcgtacatcaccattaagaccTCTctggcttgcctaaccagtgctgacaatacaaccgttcaggctgtagcaagcgcaatcggtcggACCattgaggacgaggctcgcttcggtcgtatccgtgaccttgaagctaagcacttcaagaaaaacgttgaggaacaactcaacaagcgcgtagggcacgtctacaagGGTGGTGGCGGTAGCGGAGGCGGTGGTAGCGGTGGTGGCGGTAGCGAAGTTCAGCTGCAAGCAAGCGGTGGTGGTCTGGTTCAGGCAGGCGGTAGCCTGCGTCTGAGCTGTACCGCAAGCGGTCGTACCGGCACCATTTATAGCATGGCATGGTTTCGTCAGGCACCGGGTAAAGAACGTGAATTTCTGGCAACCGTTGGTTGGAGCAGCGGTATTACCTATTATATGGATAGCGTGAAAGGTCGCTTTACCATTAGCCGTGATAATGCAAAAAATAGCGCCTACCTGCAAATGAATAGCCTGAAACCGGAAGATACCGCAGTGTATTATTGTACAGCAACCCGTGCATATAGCGTGGGTTATGATTATTGGGGTCAGGGCACCCAGGTTACCGTTAGCCATtaatactagagaacgcatgagAAAGCCCCCGGAAGATCACCTTCCGGGGGCTTTtttattgcgctactagtagcggccgctgcaggcatgcaagcttgcggccgcgtcgtgactgggaaaaccctggcgactagtcttggactcctgttgatagatccagtaatgacctcagaactccatctggatttgttcagaacgctcggttgccgccgggcgttttttattggtgagaatccaggggtccccaataattacgatttaaattggcgaaaatgagacgttgatcggcacgtaagaggttccaactttcaccataatgaaataagatcactaccgggcgtattttttgagttatcgagattttcaggagctaaggaagctaaaatggagaaaaaaatcactggatataccaccgttgatatatcccaatggcatcgtaaagaacattttgaggcatttcagtcagttgctcaatgtacctataaccagaccgttcagctggatattacggcctttttaaagaccgtaaagaaaaataagcacaagttttatccggcctttattcacattcttgcccgcctgatgaatgctcatccggaatttcgtatggcaatgaaagacggtgagctggtgatatgggatagtgttcacccttgttacaccgttttccatgagcaaactgaaacgttttcatcgctctggagtgaataccacgacgatttccggcagtttctacacatatattcgcaagatgtggcgtgttacggtgaaaacctggcctatttccctaaagggtttattgagaatatgtttttcgtctcagccaatccctgggtgagtttcaccagttttgatttaaacgtggccaatatggacaacttcttcgcccccgttttcaccatgggcaaatattatacgcaaggcgacaaggtgctgatgccgctggcgattcaggttcatcatgccgtttgtgatggcttccatgtcggcagaatgcttaatgaattacaacagtactgcgatgagtggcagggcggggcgtaatttgacttttgtccttttccgctgcataaccctgcttcggggtcattatagcgattttttcggtatatccatcctttttcgcacgatatacaggattttgccaaagggttcgtgtagactttccttggtgtatccaacggcgtcagccgggcaggataggtgaagtaggcccacccgcgagcgggtgttccttcttcactgtcccttattcgcacctggcggtgctcaacgggaatcctgctctgcgaggctggccgtaggccggccgcgatgcaggtggctgctgaacccccagccggaactgaccccacaaggccctagcgtttgcaatgcaccaggtcatcattgacccaggcgtgttccaccaggccgctgcctcgcaactcttcgcaggcttcgccgacctgctcgcgccacttcttcacgcgggtggaatccgatccgcacatgaggcggaaggtttccagcttgagcgggtacggctcccggtgcgagctgaaatagtcgaacatccgtcgggccgtcggcgacagcttgcggtacttctcccatatgaatttcgtgtagtggtcgccagcaaacagcacgacgatttcctcgtcgatcaggacctggcaacgggacgttttcttgccacggtccaggacgcggaagcggtgcagcagcgacaccgattccaggtgcccaacgcggtcggacgtgaagcccatcgccgtcgcctgtaggcgcgacaggcattcctcggccttcgtgtaataccggccattgatcgaccagcccaggtcctggcaaagctcgtagaacgtgaaggtgatcggctcgccgataggggtgcgcttcgcgtactccaacacctgctgccacaccagttcgtcatcgtcggcccgcagctcgacgccggtgtaggtgatcttcacgtccttgttgacgtggaaaatgaccttgttttgcagcgcctcgcgcgggattttcttgttgcgcgtggtgaacagggcagagcgggccgtgtcgtttggcatcgctcgcatcgtgtccggccacggcgcaatatcgaacaaggaaagctgcatttccttgatctgctgcttcgtgtgtttcagcaacgcggcctgcttggcttcgctgacctgttttgccaggtcctcgccggcggtttttcgcttcttggtcgtcatagttcctcgcgtgtcgatggtcatcgacttcgccaaacctgccgcctcctgttcgagacgacgcgaacgctccacggcggccgatggcgcgggcagggcagggggagccagttgcacgctgtcgcgctcgatcttggccgtagcttgctggactatcgagccgacggactggaaggtttcgcggggcgcacgcatgacggtgcggcttgcgatggtttcggcatcctcggcggaaaaccccgcgtcgatcagttcttgcctgtatgccttccggtcaaacgtccgattcattcaccctccttgcgggattgccccggaattaattccccggatcgatccgtcgatcttgatcccctgcgccatcagatccttggcggcaagaaagccatccagtttactttgcagggcttcccaaccttaccagagggcgccccagctggcaattccggttcgcttgctgtccataaaaccgcccagtctagctatcgccatgtaagcccactgcaagctacctgctttctctttgcgcttgcgttttcccttgtccagatagcccagtagctgacattcatccggggtcagcaccgtttctgcggactggctttctacgtggctgccatttttggggtgaggccgttcgcggccgaggggcgcagcccctggggggatgggaggcccgcgttagcgggccgggagggttcgagaagggggggcaccccccttcggcgtgcgcggtcacgcgcacagggcgcagccctggttaaaaacaaggtttataaatattggtttaaaagcaggttaaaagacaggttagcggtggccgaaaaacgggcggaaacccttgcaaatgctggattttctgcctgtggacagcccctcaaatgtcaataggtgcgcccctcatctgtcagcactctgcccctcaagtgtcaaggatcgcgcccctcatctgtcagtagtcgcgcccctcaagtgtcaataccgcagggcacttatccccaggcttgtccacatcatctgtgggaaactcgcgtaaaatcaggcgttttcgccgatttgcgaggctggccagctccacgtcgccggccgaaatcgagcctgcccctcatctgtcaacgccgcgccgggtgagtcggcccctcaagtgtcaacgtccgcccctcatctgtcagtgagggccaagttttccgcgaggtatccacaacgccggcggccctacatggctctgctgtagtgagtgggttgcgctccggcagcggtcctgatcccccgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggcgcgcccagctgtctagggcggcggatttgtcctactcaggagagcgttcaccgacaaacaacagataaaacgaaaggcccagtctttcgactgagcctttcgttttatttgatgcct