IBMc188 1 pSB3T5(BsaI-)-PBAD(SapI-)-B33-ECF20(1-101)-2aa-acVHH-T-cat-PBAD(SapI-)-RiboJ-BCD22-acVHH-2aa-ECF20(102-193)-T Creator: Trevor Y. H. Ho Principal Investigator: Baojun Wang BioSafety Level: Level 1 Backbone: pSB3T5 Origin of replication: p15A Selection Markers: Tetracycline/ Chloramphenicol pSB3T5(BsaI-)-PBAD(SapI-)-B33-ECF20(1-101)-2aa-acVHH-T-cat-PBAD(SapI-)-RiboJ-BCD22-acVHH-2aa-ECF20(102-193)-T Expression plasmid for ECF20 spit at 101/102 fused with acVHH via 2 aa linker Trevor Y. H. Ho Baojun Wang 18-NOV-2020 ds-DNA annotation4 1 cat (BsmBI-) #ff9ccd #ff9ccd cat (BsmBI-) range 1 3591 4250 annotation3 1 RFC10 Suffix #9eafd2 #9eafd2 RFC10 Suffix range 1 5588 5608 annotation0 1 VR #b4abac #b4abac VR range 1 5703 5722 annotation1 1 cat promoter #c6c9d1 #c6c9d1 cat promoter range 1 4251 4348 annotation15 1 RBS for CDS 1 #ffef86 #ffef86 RBS for CDS 1 range 1 4816 4823 annotation7 1 ECK120033737 #d59687 #d59687 ECK120033737 range 1 3506 3562 annotation23 1 acVHH #d59687 #d59687 acVHH range 1 4885 5241 annotation13 1 RBS for 2nd CDS #85dae9 #85dae9 RBS for 2nd CDS range 1 4867 4875 annotation17 1 RFC10 Prefix (full) #d6b295 #d6b295 RFC10 Prefix (full) range 1 1494 1515 annotation5 1 ECK120033736 #d59687 #d59687 ECK120033736 range 1 5535 5587 annotation9 1 tetR promoter #f58a5e #f58a5e tetR promoter range 1 9 97 annotation26 1 L3S2P55 #ff0000 #ff0000 L3S2P55 range 1 4357 4413 annotation8 1 ECF20 102-193 #ffef86 #ffef86 ECF20 102-193 range 1 5248 5523 annotation16 1 BBa_B0033 #ff9ccd #ff9ccd BBa_B0033 range 1 2714 2724 annotation6 1 VF2 #75c6a9 #75c6a9 VF2 range 1 1371 1390 annotation24 1 PBAD(SapI-) #faac61 #faac61 PBAD(SapI-) range 1 4433 4716 annotation25 1 acVHH #d59687 #d59687 acVHH range 1 3040 3396 annotation22 1 p15A origin of replication #b4abac #b4abac p15A origin of replication range 1 6169 6993 annotation19 1 PBAD(SapI-) #faac61 #faac61 PBAD(SapI-) range 1 2422 2705 annotation12 1 GG linker #b1ff67 #b1ff67 GG linker range 1 3034 3039 annotation18 1 BCD22 #faac61 #faac61 BCD22 range 1 4797 4884 annotation2 1 araC #75c6a9 #75c6a9 araC range 1 1516 2394 annotation10 1 ECK120029600 #f58a5e #f58a5e ECK120029600 range 1 3408 3497 annotation21 1 tetR #9eafd2 #9eafd2 tetR range 1 98 1291 annotation14 1 ECF20 1-101 #ffef86 #ffef86 ECF20 1-101 range 1 2731 3033 annotation20 1 RiboJ #b1ff67 #b1ff67 RiboJ insulator range 1 4722 4796 annotation11 1 GG linker #ff9ccd #ff9ccd GG linker range 1 5242 5247 IBMc188_seq 1 tactagaggagattctcatgtttgacagcttatcatcgataagctttaatgcggtagtttatcacagttaaattgctaacgcagtcaggcaccgtgtatgaaatctaacaatgcgctcatcgtcattctcggcaccgtcaccctggacgctgtaggcataggcttggttatgccggtactgccgggcctcttgcgggatatcgtccattccgacagtattgccagtcactatggcgtgctgcttgcgctctatgcgttgatgcaatttctttgcgcacccgttctcggagccctgtccgaccgctttggccgccgtccagtcctgctcgcttcgctccttggagccactatcgactacgcgatcatggcgaccacacccgtcctgtggattctctacgccggacgcatcgtggcgggcatcacgggtgccacaggtgcggttgctggtgcctatatcgccgacatcaccgatggggaagatcgggctcgccacttcgggctcatgagcgcttgtttcggcgtgggtatggtggcaggccccgtggccgggggactgttgggtgccatctccttgcatgcaccattccttgcggcggcggtgctcaacggcctcaacctcctcctgggctgcttccttatgcaggaatcgcataagggagagcgccgtccgatgcccttgcgtgccttcaatccagtcagctccttccggtgggcgcggggcatgactatcgtcgccgcacttatgactgttttctttatcatgcaactcgtaggacaggttccggcagcgctctgggtcattttcggcgaggaccgctttcgctggagcgcgacgatgatcggcctgtcgcttgcggtattcggaatcttgcacgccctcgctcaagccttcgtcacgggccccgccaccaaacgtttcggcgagaagcaggccattatcgcgggcatggcggccgacgcgctgggctacgtcttgctggcgttcgcgacgcgcggctggatggccttccccattatgattcttctcgcttccggcggcatcggtatgcccgcgttgcaggccatgctgtcccgccaagtagatgacgaccatcagggacagcttcaagggtcgctcgcggctcttaccagcctcacttcgatcattggaccgctgatcgtcacggcgatttatgccgcctcggcgagcacatggaacgggttggcatggattgtaggtgccgccctttaccttgtctgcctccccgcgttgcgtcgcggtgcatggagccgggccacctcgacctaataatactagctccggcaaaaaaacgggcaaggtgtcaccaccctgccctttttctttaaaaccgaaaagattacttcgcgtttgccacctgacgtctaagaaaaggaatattcagcaatttgcccgtgccgaagaaaggcccacccgtgaaggtgagccagtgagttgattgctacgtaattagttagttagcccttagtgactcgaattcgcggccgcttctagagttatgacaacttgacggctacatcattcactttttcttcacaaccggcacggaactcgctcgggctggccccggtgcattttttaaatacccgcgagaaatagagttgatcgtcaaaaccaacattgcgaccgacggtggcgataggcatccgggtggtgctcaaaagcagcttcgcctggctgatacgttggtcctcgcgccagcttaagacgctaatccctaactgctggcggaaaagatgtgacagacgcgacggcgacaagcaaacatgctgtgcgacgctggcgatatcaaaattgctgtctgccaggtgatcgctgatgtactgacaagcctcgcgtacccgattatccatcggtggatggagcgactcgttaatcgcttccatgcgccgcagtaacaattgctcaagcagatttatcgccagcagctccgaatagcgcccttccccttgcccggcgttaatgatttgcccaaacaggtcgctgaaatgcggctggtgcgcttcatccgggcgaaagaaccccgtattggcaaatattgacggccagttaagccattcatgccagtaggcgcgcggacgaaagtaaacccactggtgataccattcgcgagcctccggatgacgaccgtagtgatgaatctctcctggcgggaacagcaaaatatcacccggtcggcaaacaaattctcgtccctgatttttcaccaccccctgaccgcgaatggtgagattgagaatataacctttcattcccagcggtcggtcgataaaaaaatcgagataaccgttggcctcaatcggcgttaaacccgccaccagatgggcattaaacgagtatcccggcagcaggggatcattttgcgcttcagccatacttttcatactcccgccattcagagaagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggcActtctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccattactagagtcacacaggactactagatgaatgaaaccgatcctgatctggaactgctgaaacgtattggtaataatgatgcacaggccgttaaagaaatggttacccgtaaactgcctcgtctgctggcactggcaagtcgcctgctgggtgatgcagatgaagcacgtgatattgcacaagaaagttttctgcgcatttggaaacaggcagcaagctggcgtagcgaacaggcacgttttgatacctggctgcatcgtgttgcactgaatctgtgttatgatcgtctgcgtcgtcgtaaagaacatgtgccggttgatagcgaacatGGAGGTGAAGTTCAGCTGCAAGCAAGCGGTGGTGGTCTGGTTCAGGCAGGCGGTAGCCTGCGTCTGAGCTGTACCGCAAGCGGTCGTACCGGCACCATTTATAGCATGGCATGGTTTCGTCAGGCACCGGGTAAAGAACGTGAATTTCTGGCAACCGTTGGTTGGAGCAGCGGTATTACCTATTATATGGATAGCGTGAAAGGTCGCTTTACCATTAGCCGTGATAATGCAAAAAATAGCGCCTACCTGCAAATGAATAGCCTGAAACCGGAAGATACCGCAGTGTATTATTGTACAGCAACCCGTGCATATAGCGTGGGTTATGATTATTGGGGTCAGGGCACCCAGGTTACCGTTAGCCATtaaTACTAGAGttgagaagagaaaagaaaaccgccgatcctgtccaccgcattactgcaaggtagtggacaagaccggcggtcttaagttttttggctgaaTACTAGAGcctttggtcgAAAAAAAAAGCCCGCACTGTCAGGTGCGGGCTTTTTTctgtgtttccTACTAGAGCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAAACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCTACTAGAGCTCGGTACCAAAGACGAACAATAAGACGCTGAAAAGCGTCTTTTTTCGTTTTGGTCCgcttaacgatcgttggctgagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggcActtctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccatacccgagctgtcaccggatgtgctttccggtctgatgagtccgtgaggacgaaacagcctctacaaataattttgtttaaGGGCCCAAGTTCACTTAAAAAGGAGATCAACAATGAAAGCAATTTTCGTACTGAAACATCTTAATCATGCCTAGGAAGTTTTCTAATGGAAGTTCAGCTGCAAGCAAGCGGTGGTGGTCTGGTTCAGGCAGGCGGTAGCCTGCGTCTGAGCTGTACCGCAAGCGGTCGTACCGGCACCATTTATAGCATGGCATGGTTTCGTCAGGCACCGGGTAAAGAACGTGAATTTCTGGCAACCGTTGGTTGGAGCAGCGGTATTACCTATTATATGGATAGCGTGAAAGGTCGCTTTACCATTAGCCGTGATAATGCAAAAAATAGCGCCTACCTGCAAATGAATAGCCTGAAACCGGAAGATACCGCAGTGTATTATTGTACAGCAACCCGTGCATATAGCGTGGGTTATGATTATTGGGGTCAGGGCACCCAGGTTACCGTTAGCCATGGTTCAgcctgtgaagcactggatacccgtccggcaccggatgaacagctggaagcaagcgcacagagccgtcgtatggcacaggcactggatcagctgccggatcgtcagcgtgaagcaattgttctgcagtattatcaagaactgagcaataccgaagcagcagcactgatgcaaattagcgttgaagccctggaaagcctgctgagccgtgcacgtcgtaatctgcgtagccatctggccgaagcaccgggtgcagatctgagcggtcgtcgcaaaccgtaatactagagaacgcatgagAAAGCCCCCGGAAGATCACCTTCCGGGGGCTTTtttattgcgctactagtagcggccgctgcaggagtcactaagggttagttagttagattagcagaaagtcaaaagcctccgaccggaggcttttgactaaaacttcccttggggttatcattggggctcactcaaaggcggtaatcagataaaaaaaatccttagctttcgctaaggatgatttctgctagagatggaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtggatctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccttaataagatgatcttcttgagatcgttttggtctgcgcgtaatctcttgctctgaaaacgaaaaaaccgccttgcagggcggtttttcgaaggttctctgagctaccaactctttgaaccgaggtaactggcttggaggagcgcagtcaccaaaacttgtcctttcagtttagccttaaccggcgcatgacttcaagactaactcctctaaatcaattaccagtggctgctgccagtggtgcttttgcatgtctttccgggttggactcaagacgatagttaccggataaggcgcagcggtcggactgaacggggggttcgtgcatacagtccagcttggagcgaactgcctacccggaactgagtgtcaggcgtggaatgagacaaacgcggccataacagcggaatgacaccggtaaaccgaaaggcaggaacaggagagcgcacgagggagccgccaggggaaacgcctggtatctttatagtcctgtcgggtttcgccaccactgatttgagcgtcagatttcgtgatgcttgtcaggggggcggagcctatggaaaaacggctttgccgcggccctctcacttccctgttaagtatcttcctggcatcttccaggaaatctccgccccgttcgtaagccatttccgctcgccgcagtcgaacgaccgagcgtagcgagtcagtgagcgaggaagcggaatatatcctgtatcacatattctgctgacgcaccggtgcagccttttttctcctgccacatgaagcacttcactgacaccctcatcagtgccaacatagtaagccagtatacactccgctagcgctgaggtctgcctcgtgaagaaggtgttgctgactcataccaggcctgaatcgccccatcatccagccagaaagtgagggagccacggttgatgagagctttgttgtaggtggaccagttggtgattttgaacttttgctttgccacggaacggtctgcgttgtcgggaagatgcgtgatctgatccttcaactcagcaaaagttcgatttattcaacaaagccacgttgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgtt