IBMc335 1 pSB3T5(BsaI-)-PBAD(SapI-)-B30-mCherry*(1-192)-GAEY-e-NpuNcage-acVHH-T-cat-PBAD(SapI-)-RiboJ-BCD1-acVHH-NpuCcage-CFNS-mCherry*(193-236)-H6-Ter ds-DNA pSB3T5(BsaI-)-PBAD(SapI-)-B30-mCherry*(1-192)-GAEY-e-NpuNcage-acVHH-T-cat-PBAD(SapI-)-RiboJ-BCD1-acVHH-NpuCcage-CFNS-mCherry*(193-236)-H6-Ter Expression plasmid for mCherry split at 192/193 inserted with Npu DnaE, caged intein fused with acVHH Creator: Trevor Y. H. Ho Principal Investigator: Baojun Wang BioSafety Level: Level 1 Backbone: pSB3T5 Origin of replication: p15A Selection Markers: Tetracycline/ Chloramphenicol 18-NOV-2020 Trevor Y. H. Ho Baojun Wang annotation10 1 BCD1 #008000 #008000 BCD1 range 1 5641 5725 annotation5 1 Caspase3 cut site #b7e6d7 #b7e6d7 Caspase3 cut site range 1 6272 6283 annotation22 1 PBAD(SapI-) #faac61 #faac61 PBAD(SapI-) range 1 5277 5560 annotation20 1 VR #b4abac #b4abac VR range 1 6745 6764 annotation1 1 ECK120033737 #d59687 #d59687 ECK120033737 range 1 4350 4406 annotation27 1 NpuN(E52K,E54K,D85K,E89R,E91K) #ffef86 #ffef86 NpuN(E52K,E54K,D85K,E89R,E91K) range 1 3323 3628 annotation2 1 araC #75c6a9 #75c6a9 araC range 1 1516 2394 annotation17 1 PBAD(SapI-) #faac61 #faac61 PBAD(SapI-) range 1 2422 2705 annotation8 1 Caspase3 cut site #85dae9 #85dae9 Caspase3 cut site range 1 3644 3655 annotation26 1 ECK120033736 #d59687 #d59687 ECK120033736 range 1 6577 6629 annotation28 1 cat promoter #c6c9d1 #c6c9d1 cat promoter range 1 5095 5192 annotation21 1 RFC10 Prefix (full) #d6b295 #d6b295 RFC10 Prefix (full) range 1 1494 1515 annotation14 1 mCherry*(1-192) #faac61 #faac61 mCherry*(1-192) range 1 2735 3309 annotation9 1 NpuN(51-102) #f58a5e #f58a5e NpuN(51-102) range 1 6101 6256 annotation19 1 RiboJ #b1ff67 #b1ff67 RiboJ insulator range 1 5566 5640 annotation31 1 BBa_B0030 #c7b0e3 #c7b0e3 BBa_B0030 range 1 2714 2728 annotation30 1 cat (BsmBI-) #ff9ccd #ff9ccd cat (BsmBI-) range 1 4435 5094 annotation15 1 acVHH #d59687 #d59687 acVHH range 1 3884 4240 annotation0 1 mCherry*(193-236) #faac61 #faac61 mCherry*(193-236) range 1 6416 6547 annotation24 1 L3S2P55 #ff0000 #ff0000 L3S2P55 range 1 5201 5257 annotation12 1 acVHH #d59687 #d59687 acVHH range 1 5729 6085 annotation18 1 p15A origin of replication #b4abac #b4abac p15A origin of replication range 1 7211 8035 annotation25 1 tetR #9eafd2 #9eafd2 tetR range 1 98 1291 annotation29 1 H6 #c6c9d1 #c6c9d1 H6 range 1 6548 6565 annotation16 1 RBS for CDS 1 #ffef86 #ffef86 RBS for CDS 1 range 1 5660 5667 annotation6 1 VF2 #75c6a9 #75c6a9 VF2 range 1 1371 1390 annotation23 1 RFC10 Suffix #9eafd2 #9eafd2 RFC10 Suffix range 1 6630 6650 annotation4 1 ECK120029600 #f58a5e #f58a5e ECK120029600 range 1 4252 4341 annotation11 1 tetR promoter #f58a5e #f58a5e tetR promoter range 1 9 97 annotation7 1 (NpuC1-30(K2E,R6E,K11E))x2 #d6b295 #d6b295 (NpuC1-30(K2E,R6E,K11E))x2 range 1 3680 3862 annotation13 1 RBS for CDS2 #b4abac #b4abac RBS for CDS2 range 1 5711 5719 annotation3 1 NpuC #f8d3a9 #f8d3a9 NpuC range 1 6299 6412 IBMc335_seq 1 tactagaggagattctcatgtttgacagcttatcatcgataagctttaatgcggtagtttatcacagttaaattgctaacgcagtcaggcaccgtgtatgaaatctaacaatgcgctcatcgtcattctcggcaccgtcaccctggacgctgtaggcataggcttggttatgccggtactgccgggcctcttgcgggatatcgtccattccgacagtattgccagtcactatggcgtgctgcttgcgctctatgcgttgatgcaatttctttgcgcacccgttctcggagccctgtccgaccgctttggccgccgtccagtcctgctcgcttcgctccttggagccactatcgactacgcgatcatggcgaccacacccgtcctgtggattctctacgccggacgcatcgtggcgggcatcacgggtgccacaggtgcggttgctggtgcctatatcgccgacatcaccgatggggaagatcgggctcgccacttcgggctcatgagcgcttgtttcggcgtgggtatggtggcaggccccgtggccgggggactgttgggtgccatctccttgcatgcaccattccttgcggcggcggtgctcaacggcctcaacctcctcctgggctgcttccttatgcaggaatcgcataagggagagcgccgtccgatgcccttgcgtgccttcaatccagtcagctccttccggtgggcgcggggcatgactatcgtcgccgcacttatgactgttttctttatcatgcaactcgtaggacaggttccggcagcgctctgggtcattttcggcgaggaccgctttcgctggagcgcgacgatgatcggcctgtcgcttgcggtattcggaatcttgcacgccctcgctcaagccttcgtcacgggccccgccaccaaacgtttcggcgagaagcaggccattatcgcgggcatggcggccgacgcgctgggctacgtcttgctggcgttcgcgacgcgcggctggatggccttccccattatgattcttctcgcttccggcggcatcggtatgcccgcgttgcaggccatgctgtcccgccaagtagatgacgaccatcagggacagcttcaagggtcgctcgcggctcttaccagcctcacttcgatcattggaccgctgatcgtcacggcgatttatgccgcctcggcgagcacatggaacgggttggcatggattgtaggtgccgccctttaccttgtctgcctccccgcgttgcgtcgcggtgcatggagccgggccacctcgacctaataatactagctccggcaaaaaaacgggcaaggtgtcaccaccctgccctttttctttaaaaccgaaaagattacttcgcgtttgccacctgacgtctaagaaaaggaatattcagcaatttgcccgtgccgaagaaaggcccacccgtgaaggtgagccagtgagttgattgctacgtaattagttagttagcccttagtgactcgaattcgcggccgcttctagagttatgacaacttgacggctacatcattcactttttcttcacaaccggcacggaactcgctcgggctggccccggtgcattttttaaatacccgcgagaaatagagttgatcgtcaaaaccaacattgcgaccgacggtggcgataggcatccgggtggtgctcaaaagcagcttcgcctggctgatacgttggtcctcgcgccagcttaagacgctaatccctaactgctggcggaaaagatgtgacagacgcgacggcgacaagcaaacatgctgtgcgacgctggcgatatcaaaattgctgtctgccaggtgatcgctgatgtactgacaagcctcgcgtacccgattatccatcggtggatggagcgactcgttaatcgcttccatgcgccgcagtaacaattgctcaagcagatttatcgccagcagctccgaatagcgcccttccccttgcccggcgttaatgatttgcccaaacaggtcgctgaaatgcggctggtgcgcttcatccgggcgaaagaaccccgtattggcaaatattgacggccagttaagccattcatgccagtaggcgcgcggacgaaagtaaacccactggtgataccattcgcgagcctccggatgacgaccgtagtgatgaatctctcctggcgggaacagcaaaatatcacccggtcggcaaacaaattctcgtccctgatttttcaccaccccctgaccgcgaatggtgagattgagaatataacctttcattcccagcggtcggtcgataaaaaaatcgagataaccgttggcctcaatcggcgttaaacccgccaccagatgggcattaaacgagtatcccggcagcaggggatcattttgcgcttcagccatacttttcatactcccgccattcagagaagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggcActtctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccattactagagattaaagaggagaaatactagatggtgagcaagggcgaagaaGATaacatggccatcatcaaggagttcatgcgcttcaaggtgcacatggagggctccgtgaacggccacgagttcgagatcgagggcgagggcgagggccgcccctacgagggcacccagaccgccaagctgaaggtgaccaagggtggccccctgcccttcgcctgggacatcctgtcccctcagttcatgtacggctccaaggcctacgtgaagcaccccgccgacatccccgactacttgaagctgtccttccccgagggcttcaagtgggagcgcgtgatgaacttcgaggacggcggcgtggtgaccgtgacccaggactcctccttgcaggacggcgagttcatctacaaggtgaagctgcgcggcaccaacttcccctccgacggccccgtaatgcagaagAAAaccatgggctgggaggcctcctccgagcggatgtaccccgaggacggcgccctgaagggcgagatcaagcagaggctgaagctgaaggacggcggccactacgacgctgaggtcaagaccacctacaaggccaagaagcccGTTGGAGCAGAATATTGCCTGAGCTATGAAACCGAAATTCTGACCGTTGAATATGGTCTGCTGCCGATTGGTAAAATTGTGGAAAAACGTATTGAGTGCACCGTGTATAGCGTGGATAATAACGGTAACATTTATACCCAGCCGGTTGCACAGTGGCATGATCGTGGTAAACAGAAAGTTTTTGAGTACTGTCTGGAAGATGGTAGCCTGATTCGTGCAACCAAAGATCACAAATTTATGACCGTGGATGGTCAGATGCTGCCCATTAAAGAAATTTTTCGTCGTAAACTGGATCTGATGCGTGTTGATAATCTGCCGAATGAAAATCTGTATGGTGATGAAGTGGATGGCTATTTTCAGGGTATTGAAATTGCCACCGAAAAATATCTGGGTGAGCAGAACGTTTATGATATTGGTGTTGAACGCGATCATAACTTCGCACTGAAAAATGGTGGTTATTTCCAGGGCATCGAAATCGCAACAGAGAAATACCTGGGCGAACAGAATGTGTATGACATCGGCGTGGAACGTGATCACAATTTTGCCCTGAAGAATGGCGGTAGCGGTAGCGCAAGCGAAGTTCAGCTGCAAGCAAGCGGTGGTGGTCTGGTTCAGGCAGGCGGTAGCCTGCGTCTGAGCTGTACCGCAAGCGGTCGTACCGGCACCATTTATAGCATGGCATGGTTTCGTCAGGCACCGGGTAAAGAACGTGAATTTCTGGCAACCGTTGGTTGGAGCAGCGGTATTACCTATTATATGGATAGCGTGAAAGGTCGCTTTACCATTAGCCGTGATAATGCAAAAAATAGCGCCTACCTGCAAATGAATAGCCTGAAACCGGAAGATACCGCAGTGTATTATTGTACAGCAACCCGTGCATATAGCGTGGGTTATGATTATTGGGGTCAGGGCACCCAGGTTACCGTTAGCCATtaaTACTAGAGttgagaagagaaaagaaaaccgccgatcctgtccaccgcattactgcaaggtagtggacaagaccggcggtcttaagttttttggctgaaTACTAGAGcctttggtcgAAAAAAAAAGCCCGCACTGTCAGGTGCGGGCTTTTTTctgtgtttccTACTAGAGCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAAACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCTACTAGAGCTCGGTACCAAAGACGAACAATAAGACGCTGAAAAGCGTCTTTTTTCGTTTTGGTCCgcttaacgatcgttggctgagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggcActtctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccatacccgagctgtcaccggatgtgctttccggtctgatgagtccgtgaggacgaaacagcctctacaaataattttgtttaaGGGCCCAAGTTCACTTAAAAAGGAGATCAACAATGAAAGCAATTTTCGTACTGAAACATCTTAATCATGCACAGGAGACTTTCTAatgGAAGTTCAGCTGCAAGCAAGCGGTGGTGGTCTGGTTCAGGCAGGCGGTAGCCTGCGTCTGAGCTGTACCGCAAGCGGTCGTACCGGCACCATTTATAGCATGGCATGGTTTCGTCAGGCACCGGGTAAAGAACGTGAATTTCTGGCAACCGTTGGTTGGAGCAGCGGTATTACCTATTATATGGATAGCGTGAAAGGTCGCTTTACCATTAGCCGTGATAATGCAAAAAATAGCGCCTACCTGCAAATGAATAGCCTGAAACCGGAAGATACCGCAGTGTATTATTGTACAGCAACCCGTGCATATAGCGTGGGTTATGATTATTGGGGTCAGGGCACCCAGGTTACCGTTAGCCATGGTAATGGTAATGGTGGTGAACAAGAAGTGTTTGAGTATTGTCTGGAAGATGGTAGCCTGATTCGTGCAACCAAAGATCACAAATTTATGACCGTTGATGGTCAGATGCTGCCGATTGATGAAATTTTTGAACGTGAACTGGATCTGATGCGTGTTGATAATCTGCCGAATGAAAATCTGTATGGTGATGAAGTGGATGGCTATTTTCAGGGTATCAAAATTGCGACCCGTAAATATCTGGGTAAACAGAACGTTTATGATATTGGTGTGGAACGCGATCATAACTTTGCACTGAAAAATGGTTTTATCGCCAGCAACTGCTTCAATTCAcagctgcccggcgcctacaacgtcaacatcaagttggacatcacctcccacaacgaggactacaccatcgtggaacagtacgaacgcgccgagggccgccactccaccggcggcatggacgagctgtacaagcatcatcaccatcaccactaatactagagaacgcatgagAAAGCCCCCGGAAGATCACCTTCCGGGGGCTTTtttattgcgctactagtagcggccgctgcaggagtcactaagggttagttagttagattagcagaaagtcaaaagcctccgaccggaggcttttgactaaaacttcccttggggttatcattggggctcactcaaaggcggtaatcagataaaaaaaatccttagctttcgctaaggatgatttctgctagagatggaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtggatctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccttaataagatgatcttcttgagatcgttttggtctgcgcgtaatctcttgctctgaaaacgaaaaaaccgccttgcagggcggtttttcgaaggttctctgagctaccaactctttgaaccgaggtaactggcttggaggagcgcagtcaccaaaacttgtcctttcagtttagccttaaccggcgcatgacttcaagactaactcctctaaatcaattaccagtggctgctgccagtggtgcttttgcatgtctttccgggttggactcaagacgatagttaccggataaggcgcagcggtcggactgaacggggggttcgtgcatacagtccagcttggagcgaactgcctacccggaactgagtgtcaggcgtggaatgagacaaacgcggccataacagcggaatgacaccggtaaaccgaaaggcaggaacaggagagcgcacgagggagccgccaggggaaacgcctggtatctttatagtcctgtcgggtttcgccaccactgatttgagcgtcagatttcgtgatgcttgtcaggggggcggagcctatggaaaaacggctttgccgcggccctctcacttccctgttaagtatcttcctggcatcttccaggaaatctccgccccgttcgtaagccatttccgctcgccgcagtcgaacgaccgagcgtagcgagtcagtgagcgaggaagcggaatatatcctgtatcacatattctgctgacgcaccggtgcagccttttttctcctgccacatgaagcacttcactgacaccctcatcagtgccaacatagtaagccagtatacactccgctagcgctgaggtctgcctcgtgaagaaggtgttgctgactcataccaggcctgaatcgccccatcatccagccagaaagtgagggagccacggttgatgagagctttgttgtaggtggaccagttggtgattttgaacttttgctttgccacggaacggtctgcgttgtcgggaagatgcgtgatctgatccttcaactcagcaaaagttcgatttattcaacaaagccacgttgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgtt