IBMc191 1 pSB3T5(BsaI-)-PBAD(SapI-)-B33-ECF20(1-101)-15aa-acVHH-T-cat-PBAD(SapI-)-RiboJ-BCD22-acVHH-15aa-ECF20(102-193)-T ds-DNA pSB3T5(BsaI-)-PBAD(SapI-)-B33-ECF20(1-101)-15aa-acVHH-T-cat-PBAD(SapI-)-RiboJ-BCD22-acVHH-15aa-ECF20(102-193)-T Expression plasmid for ECF20 spit at 101/102 fused with acVHH via 15 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 Trevor Y. H. Ho Baojun Wang annotation7 1 3x(GGGGS) linker #b1ff67 #b1ff67 3x(GGGGS) linker range 1 3034 3078 annotation6 1 tetR promoter #f58a5e #f58a5e tetR promoter range 1 9 97 annotation4 1 ECK120029600 #f58a5e #f58a5e ECK120029600 range 1 3447 3536 annotation16 1 p15A origin of replication #b4abac #b4abac p15A origin of replication range 1 6247 7071 annotation12 1 BCD22 #faac61 #faac61 BCD22 range 1 4836 4923 annotation20 1 acVHH #d59687 #d59687 acVHH range 1 3079 3435 annotation10 1 RBS for CDS 1 #ffef86 #ffef86 RBS for CDS 1 range 1 4855 4862 annotation26 1 araC #75c6a9 #75c6a9 araC range 1 1516 2394 annotation8 1 RBS for 2nd CDS #85dae9 #85dae9 RBS for 2nd CDS range 1 4906 4914 annotation13 1 RFC10 Prefix (full) #d6b295 #d6b295 RFC10 Prefix (full) range 1 1494 1515 annotation21 1 L3S2P55 #ff0000 #ff0000 L3S2P55 range 1 4396 4452 annotation1 1 ECK120033737 #d59687 #d59687 ECK120033737 range 1 3545 3601 annotation14 1 PBAD(SapI-) #faac61 #faac61 PBAD(SapI-) range 1 2422 2705 annotation9 1 ECF20 1-101 #ffef86 #ffef86 ECF20 1-101 range 1 2731 3033 annotation15 1 RiboJ #b1ff67 #b1ff67 RiboJ insulator range 1 4761 4835 annotation5 1 3x(GGGGS) linker #ff9ccd #ff9ccd 3x(GGGGS) linker range 1 5281 5325 annotation25 1 cat (BsmBI-) #ff9ccd #ff9ccd cat (BsmBI-) range 1 3630 4289 annotation11 1 BBa_B0033 #ff9ccd #ff9ccd BBa_B0033 range 1 2714 2724 annotation0 1 ECK120033736 #d59687 #d59687 ECK120033736 range 1 5613 5665 annotation19 1 PBAD(SapI-) #faac61 #faac61 PBAD(SapI-) range 1 4472 4755 annotation23 1 cat promoter #c6c9d1 #c6c9d1 cat promoter range 1 4290 4387 annotation22 1 VR #b4abac #b4abac VR range 1 5781 5800 annotation18 1 tetR #9eafd2 #9eafd2 tetR range 1 98 1291 annotation24 1 RFC10 Suffix #9eafd2 #9eafd2 RFC10 Suffix range 1 5666 5686 annotation2 1 VF2 #75c6a9 #75c6a9 VF2 range 1 1371 1390 annotation3 1 ECF20 102-193 #ffef86 #ffef86 ECF20 102-193 range 1 5326 5601 annotation17 1 acVHH #d59687 #d59687 acVHH range 1 4924 5280 IBMc191_seq 1 tactagaggagattctcatgtttgacagcttatcatcgataagctttaatgcggtagtttatcacagttaaattgctaacgcagtcaggcaccgtgtatgaaatctaacaatgcgctcatcgtcattctcggcaccgtcaccctggacgctgtaggcataggcttggttatgccggtactgccgggcctcttgcgggatatcgtccattccgacagtattgccagtcactatggcgtgctgcttgcgctctatgcgttgatgcaatttctttgcgcacccgttctcggagccctgtccgaccgctttggccgccgtccagtcctgctcgcttcgctccttggagccactatcgactacgcgatcatggcgaccacacccgtcctgtggattctctacgccggacgcatcgtggcgggcatcacgggtgccacaggtgcggttgctggtgcctatatcgccgacatcaccgatggggaagatcgggctcgccacttcgggctcatgagcgcttgtttcggcgtgggtatggtggcaggccccgtggccgggggactgttgggtgccatctccttgcatgcaccattccttgcggcggcggtgctcaacggcctcaacctcctcctgggctgcttccttatgcaggaatcgcataagggagagcgccgtccgatgcccttgcgtgccttcaatccagtcagctccttccggtgggcgcggggcatgactatcgtcgccgcacttatgactgttttctttatcatgcaactcgtaggacaggttccggcagcgctctgggtcattttcggcgaggaccgctttcgctggagcgcgacgatgatcggcctgtcgcttgcggtattcggaatcttgcacgccctcgctcaagccttcgtcacgggccccgccaccaaacgtttcggcgagaagcaggccattatcgcgggcatggcggccgacgcgctgggctacgtcttgctggcgttcgcgacgcgcggctggatggccttccccattatgattcttctcgcttccggcggcatcggtatgcccgcgttgcaggccatgctgtcccgccaagtagatgacgaccatcagggacagcttcaagggtcgctcgcggctcttaccagcctcacttcgatcattggaccgctgatcgtcacggcgatttatgccgcctcggcgagcacatggaacgggttggcatggattgtaggtgccgccctttaccttgtctgcctccccgcgttgcgtcgcggtgcatggagccgggccacctcgacctaataatactagctccggcaaaaaaacgggcaaggtgtcaccaccctgccctttttctttaaaaccgaaaagattacttcgcgtttgccacctgacgtctaagaaaaggaatattcagcaatttgcccgtgccgaagaaaggcccacccgtgaaggtgagccagtgagttgattgctacgtaattagttagttagcccttagtgactcgaattcgcggccgcttctagagttatgacaacttgacggctacatcattcactttttcttcacaaccggcacggaactcgctcgggctggccccggtgcattttttaaatacccgcgagaaatagagttgatcgtcaaaaccaacattgcgaccgacggtggcgataggcatccgggtggtgctcaaaagcagcttcgcctggctgatacgttggtcctcgcgccagcttaagacgctaatccctaactgctggcggaaaagatgtgacagacgcgacggcgacaagcaaacatgctgtgcgacgctggcgatatcaaaattgctgtctgccaggtgatcgctgatgtactgacaagcctcgcgtacccgattatccatcggtggatggagcgactcgttaatcgcttccatgcgccgcagtaacaattgctcaagcagatttatcgccagcagctccgaatagcgcccttccccttgcccggcgttaatgatttgcccaaacaggtcgctgaaatgcggctggtgcgcttcatccgggcgaaagaaccccgtattggcaaatattgacggccagttaagccattcatgccagtaggcgcgcggacgaaagtaaacccactggtgataccattcgcgagcctccggatgacgaccgtagtgatgaatctctcctggcgggaacagcaaaatatcacccggtcggcaaacaaattctcgtccctgatttttcaccaccccctgaccgcgaatggtgagattgagaatataacctttcattcccagcggtcggtcgataaaaaaatcgagataaccgttggcctcaatcggcgttaaacccgccaccagatgggcattaaacgagtatcccggcagcaggggatcattttgcgcttcagccatacttttcatactcccgccattcagagaagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggcActtctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccattactagagtcacacaggactactagatgaatgaaaccgatcctgatctggaactgctgaaacgtattggtaataatgatgcacaggccgttaaagaaatggttacccgtaaactgcctcgtctgctggcactggcaagtcgcctgctgggtgatgcagatgaagcacgtgatattgcacaagaaagttttctgcgcatttggaaacaggcagcaagctggcgtagcgaacaggcacgttttgatacctggctgcatcgtgttgcactgaatctgtgttatgatcgtctgcgtcgtcgtaaagaacatgtgccggttgatagcgaacatGGAGGTGGCGGTAGCGGAGGCGGTGGTAGCGGTGGTGGCGGTAGCGAAGTTCAGCTGCAAGCAAGCGGTGGTGGTCTGGTTCAGGCAGGCGGTAGCCTGCGTCTGAGCTGTACCGCAAGCGGTCGTACCGGCACCATTTATAGCATGGCATGGTTTCGTCAGGCACCGGGTAAAGAACGTGAATTTCTGGCAACCGTTGGTTGGAGCAGCGGTATTACCTATTATATGGATAGCGTGAAAGGTCGCTTTACCATTAGCCGTGATAATGCAAAAAATAGCGCCTACCTGCAAATGAATAGCCTGAAACCGGAAGATACCGCAGTGTATTATTGTACAGCAACCCGTGCATATAGCGTGGGTTATGATTATTGGGGTCAGGGCACCCAGGTTACCGTTAGCCATtaaTACTAGAGttgagaagagaaaagaaaaccgccgatcctgtccaccgcattactgcaaggtagtggacaagaccggcggtcttaagttttttggctgaaTACTAGAGcctttggtcgAAAAAAAAAGCCCGCACTGTCAGGTGCGGGCTTTTTTctgtgtttccTACTAGAGCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTGTTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATCAGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGTTTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAAACGAAAAACATATTCTCAATAAACCCTTTAGGGAAATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTATTCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCACCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGCCGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAGGTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATCCAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGATCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCTACTAGAGCTCGGTACCAAAGACGAACAATAAGACGCTGAAAAGCGTCTTTTTTCGTTTTGGTCCgcttaacgatcgttggctgagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggcActtctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccatacccgagctgtcaccggatgtgctttccggtctgatgagtccgtgaggacgaaacagcctctacaaataattttgtttaaGGGCCCAAGTTCACTTAAAAAGGAGATCAACAATGAAAGCAATTTTCGTACTGAAACATCTTAATCATGCCTAGGAAGTTTTCTAATGGAAGTTCAGCTGCAAGCAAGCGGTGGTGGTCTGGTTCAGGCAGGCGGTAGCCTGCGTCTGAGCTGTACCGCAAGCGGTCGTACCGGCACCATTTATAGCATGGCATGGTTTCGTCAGGCACCGGGTAAAGAACGTGAATTTCTGGCAACCGTTGGTTGGAGCAGCGGTATTACCTATTATATGGATAGCGTGAAAGGTCGCTTTACCATTAGCCGTGATAATGCAAAAAATAGCGCCTACCTGCAAATGAATAGCCTGAAACCGGAAGATACCGCAGTGTATTATTGTACAGCAACCCGTGCATATAGCGTGGGTTATGATTATTGGGGTCAGGGCACCCAGGTTACCGTTAGCCATGGTGGAGGTGGTTCAGGTGGCGGAGGTTCTGGTGGAGGTGGTTCAgcctgtgaagcactggatacccgtccggcaccggatgaacagctggaagcaagcgcacagagccgtcgtatggcacaggcactggatcagctgccggatcgtcagcgtgaagcaattgttctgcagtattatcaagaactgagcaataccgaagcagcagcactgatgcaaattagcgttgaagccctggaaagcctgctgagccgtgcacgtcgtaatctgcgtagccatctggccgaagcaccgggtgcagatctgagcggtcgtcgcaaaccgtaatactagagaacgcatgagAAAGCCCCCGGAAGATCACCTTCCGGGGGCTTTtttattgcgctactagtagcggccgctgcaggagtcactaagggttagttagttagattagcagaaagtcaaaagcctccgaccggaggcttttgactaaaacttcccttggggttatcattggggctcactcaaaggcggtaatcagataaaaaaaatccttagctttcgctaaggatgatttctgctagagatggaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtggatctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccttaataagatgatcttcttgagatcgttttggtctgcgcgtaatctcttgctctgaaaacgaaaaaaccgccttgcagggcggtttttcgaaggttctctgagctaccaactctttgaaccgaggtaactggcttggaggagcgcagtcaccaaaacttgtcctttcagtttagccttaaccggcgcatgacttcaagactaactcctctaaatcaattaccagtggctgctgccagtggtgcttttgcatgtctttccgggttggactcaagacgatagttaccggataaggcgcagcggtcggactgaacggggggttcgtgcatacagtccagcttggagcgaactgcctacccggaactgagtgtcaggcgtggaatgagacaaacgcggccataacagcggaatgacaccggtaaaccgaaaggcaggaacaggagagcgcacgagggagccgccaggggaaacgcctggtatctttatagtcctgtcgggtttcgccaccactgatttgagcgtcagatttcgtgatgcttgtcaggggggcggagcctatggaaaaacggctttgccgcggccctctcacttccctgttaagtatcttcctggcatcttccaggaaatctccgccccgttcgtaagccatttccgctcgccgcagtcgaacgaccgagcgtagcgagtcagtgagcgaggaagcggaatatatcctgtatcacatattctgctgacgcaccggtgcagccttttttctcctgccacatgaagcacttcactgacaccctcatcagtgccaacatagtaagccagtatacactccgctagcgctgaggtctgcctcgtgaagaaggtgttgctgactcataccaggcctgaatcgccccatcatccagccagaaagtgagggagccacggttgatgagagctttgttgtaggtggaccagttggtgattttgaacttttgctttgccacggaacggtctgcgttgtcgggaagatgcgtgatctgatccttcaactcagcaaaagttcgatttattcaacaaagccacgttgtgtctcaaaatctctgatgttacattgcacaagataaaaatatatcatcatgaacaataaaactgtctgcttacataaacagtaatacaaggggtgtt