IBMc190 1 pSB3T5(BsaI-)-PBAD(SapI-)-B33-ECF20(1-101)-10aa-acVHH-T-cat-PBAD(SapI-)-RiboJ-BCD22-acVHH-10aa-ECF20(102-193)-T ds-DNA pSB3T5(BsaI-)-PBAD(SapI-)-B33-ECF20(1-101)-10aa-acVHH-T-cat-PBAD(SapI-)-RiboJ-BCD22-acVHH-10aa-ECF20(102-193)-T Expression plasmid for ECF20 spit at 101/102 fused with acVHH via 10 aa linker 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 Baojun Wang Trevor Y. H. Ho annotation4 1 tetR #9eafd2 #9eafd2 tetR range 1 98 1291 annotation0 1 RiboJ #b1ff67 #b1ff67 RiboJ insulator range 1 4746 4820 annotation20 1 RBS for 2nd CDS #85dae9 #85dae9 RBS for 2nd CDS range 1 4891 4899 annotation23 1 BBa_B0033 #ff9ccd #ff9ccd BBa_B0033 range 1 2714 2724 annotation5 1 acVHH #d59687 #d59687 acVHH range 1 3064 3420 annotation10 1 cat (BsmBI-) #ff9ccd #ff9ccd cat (BsmBI-) range 1 3615 4274 annotation11 1 araC #75c6a9 #75c6a9 araC range 1 1516 2394 annotation7 1 VR #b4abac #b4abac VR range 1 5751 5770 annotation13 1 ECK120033737 #d59687 #d59687 ECK120033737 range 1 3530 3586 annotation21 1 ECF20 1-101 #ffef86 #ffef86 ECF20 1-101 range 1 2731 3033 annotation6 1 L3S2P55 #ff0000 #ff0000 L3S2P55 range 1 4381 4437 annotation18 1 2x(GGGGS) linker #b1ff67 #b1ff67 2x(GGGGS) linker range 1 3034 3063 annotation1 1 p15A origin of replication #b4abac #b4abac p15A origin of replication range 1 6217 7041 annotation17 1 2x(GGGGS) linker #ff9ccd #ff9ccd 2x(GGGGS) linker range 1 5266 5295 annotation2 1 acVHH #d59687 #d59687 acVHH range 1 4909 5265 annotation24 1 BCD22 #faac61 #faac61 BCD22 range 1 4821 4908 annotation25 1 PBAD(SapI-) #faac61 #faac61 PBAD(SapI-) range 1 2422 2705 annotation22 1 RBS for CDS 1 #ffef86 #ffef86 RBS for CDS 1 range 1 4840 4847 annotation15 1 VF2 #75c6a9 #75c6a9 VF2 range 1 1371 1390 annotation19 1 tetR promoter #f58a5e #f58a5e tetR promoter range 1 9 97 annotation9 1 RFC10 Suffix #9eafd2 #9eafd2 RFC10 Suffix range 1 5636 5656 annotation26 1 RFC10 Prefix (full) #d6b295 #d6b295 RFC10 Prefix (full) range 1 1494 1515 annotation3 1 PBAD(SapI-) #faac61 #faac61 PBAD(SapI-) range 1 4457 4740 annotation14 1 ECF20 102-193 #ffef86 #ffef86 ECF20 102-193 range 1 5296 5571 annotation16 1 ECK120029600 #f58a5e #f58a5e ECK120029600 range 1 3432 3521 annotation12 1 ECK120033736 #d59687 #d59687 ECK120033736 range 1 5583 5635 annotation8 1 cat promoter #c6c9d1 #c6c9d1 cat promoter range 1 4275 4372 IBMc190_seq 1 tactagaggagattctcatgtttgacagcttatcatcgataagctttaatgcggtagtttatcacagttaaattgctaacgcagtcaggcaccgtgtatgaaatctaacaatgcgctcatcgtcattctcggcaccgtcaccctggacgctgtaggcataggcttggttatgccggtactgccgggcctcttgcgggatatcgtccattccgacagtattgccagtcactatggcgtgctgcttgcgctctatgcgttgatgcaatttctttgcgcacccgttctcggagccctgtccgaccgctttggccgccgtccagtcctgctcgcttcgctccttggagccactatcgactacgcgatcatggcgaccacacccgtcctgtggattctctacgccggacgcatcgtggcgggcatcacgggtgccacaggtgcggttgctggtgcctatatcgccgacatcaccgatggggaagatcgggctcgccacttcgggctcatgagcgcttgtttcggcgtgggtatggtggcaggccccgtggccgggggactgttgggtgccatctccttgcatgcaccattccttgcggcggcggtgctcaacggcctcaacctcctcctgggctgcttccttatgcaggaatcgcataagggagagcgccgtccgatgcccttgcgtgccttcaatccagtcagctccttccggtgggcgcggggcatgactatcgtcgccgcacttatgactgttttctttatcatgcaactcgtaggacaggttccggcagcgctctgggtcattttcggcgaggaccgctttcgctggagcgcgacgatgatcggcctgtcgcttgcggtattcggaatcttgcacgccctcgctcaagccttcgtcacgggccccgccaccaaacgtttcggcgagaagcaggccattatcgcgggcatggcggccgacgcgctgggctacgtcttgctggcgttcgcgacgcgcggctggatggccttccccattatgattcttctcgcttccggcggcatcggtatgcccgcgttgcaggccatgctgtcccgccaagtagatgacgaccatcagggacagcttcaagggtcgctcgcggctcttaccagcctcacttcgatcattggaccgctgatcgtcacggcgatttatgccgcctcggcgagcacatggaacgggttggcatggattgtaggtgccgccctttaccttgtctgcctccccgcgttgcgtcgcggtgcatggagccgggccacctcgacctaataatactagctccggcaaaaaaacgggcaaggtgtcaccaccctgccctttttctttaaaaccgaaaagattacttcgcgtttgccacctgacgtctaagaaaaggaatattcagcaatttgcccgtgccgaagaaaggcccacccgtgaaggtgagccagtgagttgattgctacgtaattagttagttagcccttagtgactcgaattcgcggccgcttctagagttatgacaacttgacggctacatcattcactttttcttcacaaccggcacggaactcgctcgggctggccccggtgcattttttaaatacccgcgagaaatagagttgatcgtcaaaaccaacattgcgaccgacggtggcgataggcatccgggtggtgctcaaaagcagcttcgcctggctgatacgttggtcctcgcgccagcttaagacgctaatccctaactgctggcggaaaagatgtgacagacgcgacggcgacaagcaaacatgctgtgcgacgctggcgatatcaaaattgctgtctgccaggtgatcgctgatgtactgacaagcctcgcgtacccgattatccatcggtggatggagcgactcgttaatcgcttccatgcgccgcagtaacaattgctcaagcagatttatcgccagcagctccgaatagcgcccttccccttgcccggcgttaatgatttgcccaaacaggtcgctgaaatgcggctggtgcgcttcatccgggcgaaagaaccccgtattggcaaatattgacggccagttaagccattcatgccagtaggcgcgcggacgaaagtaaacccactggtgataccattcgcgagcctccggatgacgaccgtagtgatgaatctctcctggcgggaacagcaaaatatcacccggtcggcaaacaaattctcgtccctgatttttcaccaccccctgaccgcgaatggtgagattgagaatataacctttcattcccagcggtcggtcgataaaaaaatcgagataaccgttggcctcaatcggcgttaaacccgccaccagatgggcattaaacgagtatcccggcagcaggggatcattttgcgcttcagccatacttttcatactcccgccattcagagaagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggcActtctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccattactagagtcacacaggactactagatgaatgaaaccgatcctgatctggaactgctgaaacgtattggtaataatgatgcacaggccgttaaagaaatggttacccgtaaactgcctcgtctgctggcactggcaagtcgcctgctgggtgatgcagatgaagcacgtgatattgcacaagaaagttttctgcgcatttggaaacaggcagcaagctggcgtagcgaacaggcacgttttgatacctggctgcatcgtgttgcactgaatctgtgttatgatcgtctgcgtcgtcgtaaagaacatgtgccggttgatagcgaacatGGAGGCGGTGGTAGCGGTGGTGGCGGTAGCGAAGTTCAGCTGCAAGCAAGCGGTGGTGGTCTGGTTCAGGCAGGCGGTAGCCTGCGTCTGAGCTGTACCGCAAGCGGTCGTACCGGCACCATTTATAGCATGGCATGGTTTCGTCAGGCACCGGGTAAAGAACGTGAATTTCTGGCAACCGTTGGTTGGAGCAGCGGTATTACCTATTATATGGATAGCGTGAAAGGTCGCTTTACCATTAGCCGTGATAATGCAAAAAATAGCGCCTACCTGCAAATGAATAGCCTGAAACCGGAAGATACCGCAGTGTATTATTGTACAGCAACCCGTGCATATAGCGTGGGTTATGATTATTGGGGTCAGGGCACCCAGGTTACCGTTAGCCATtaaTACTAGAGttgagaagagaaaagaaaaccgccgatcctgtccaccgcattactgcaaggtagtggacaagaccggcggtcttaagttttttggctgaaTACTAGAGcctttggtcgAAAAAAAAAGCCCGCACTGTCAGGTGCGGGCTTTTTTctgtgtttccTACTAGAGCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAAACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCTACTAGAGCTCGGTACCAAAGACGAACAATAAGACGCTGAAAAGCGTCTTTTTTCGTTTTGGTCCgcttaacgatcgttggctgagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggcActtctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccatacccgagctgtcaccggatgtgctttccggtctgatgagtccgtgaggacgaaacagcctctacaaataattttgtttaaGGGCCCAAGTTCACTTAAAAAGGAGATCAACAATGAAAGCAATTTTCGTACTGAAACATCTTAATCATGCCTAGGAAGTTTTCTAATGGAAGTTCAGCTGCAAGCAAGCGGTGGTGGTCTGGTTCAGGCAGGCGGTAGCCTGCGTCTGAGCTGTACCGCAAGCGGTCGTACCGGCACCATTTATAGCATGGCATGGTTTCGTCAGGCACCGGGTAAAGAACGTGAATTTCTGGCAACCGTTGGTTGGAGCAGCGGTATTACCTATTATATGGATAGCGTGAAAGGTCGCTTTACCATTAGCCGTGATAATGCAAAAAATAGCGCCTACCTGCAAATGAATAGCCTGAAACCGGAAGATACCGCAGTGTATTATTGTACAGCAACCCGTGCATATAGCGTGGGTTATGATTATTGGGGTCAGGGCACCCAGGTTACCGTTAGCCATGGTGGAGGTGGTTCAGGTGGCGGAGGTTCAgcctgtgaagcactggatacccgtccggcaccggatgaacagctggaagcaagcgcacagagccgtcgtatggcacaggcactggatcagctgccggatcgtcagcgtgaagcaattgttctgcagtattatcaagaactgagcaataccgaagcagcagcactgatgcaaattagcgttgaagccctggaaagcctgctgagccgtgcacgtcgtaatctgcgtagccatctggccgaagcaccgggtgcagatctgagcggtcgtcgcaaaccgtaatactagagaacgcatgagAAAGCCCCCGGAAGATCACCTTCCGGGGGCTTTtttattgcgctactagtagcggccgctgcaggagtcactaagggttagttagttagattagcagaaagtcaaaagcctccgaccggaggcttttgactaaaacttcccttggggttatcattggggctcactcaaaggcggtaatcagataaaaaaaatccttagctttcgctaaggatgatttctgctagagatggaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtggatctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccttaataagatgatcttcttgagatcgttttggtctgcgcgtaatctcttgctctgaaaacgaaaaaaccgccttgcagggcggtttttcgaaggttctctgagctaccaactctttgaaccgaggtaactggcttggaggagcgcagtcaccaaaacttgtcctttcagtttagccttaaccggcgcatgacttcaagactaactcctctaaatcaattaccagtggctgctgccagtggtgcttttgcatgtctttccgggttggactcaagacgatagttaccggataaggcgcagcggtcggactgaacggggggttcgtgcatacagtccagcttggagcgaactgcctacccggaactgagtgtcaggcgtggaatgagacaaacgcggccataacagcggaatgacaccggtaaaccgaaaggcaggaacaggagagcgcacgagggagccgccaggggaaacgcctggtatctttatagtcctgtcgggtttcgccaccactgatttgagcgtcagatttcgtgatgcttgtcaggggggcggagcctatggaaaaacggctttgccgcggccctctcacttccctgttaagtatcttcctggcatcttccaggaaatctccgccccgttcgtaagccatttccgctcgccgcagtcgaacgaccgagcgtagcgagtcagtgagcgaggaagcggaatatatcctgtatcacatattctgctgacgcaccggtgcagccttttttctcctgccacatgaagcacttcactgacaccctcatcagtgccaacatagtaagccagtatacactccgctagcgctgaggtctgcctcgtgaagaaggtgttgctgactcataccaggcctgaatcgccccatcatccagccagaaagtgagggagccacggttgatgagagctttgttgtaggtggaccagttggtgattttgaacttttgctttgccacggaacggtctgcgttgtcgggaagatgcgtgatctgatccttcaactcagcaaaagttcgatttattcaacaaagccacgttgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgtt