BBa_K1962002 1 BBa_K1962002 Truncated Colicin Ia Lacking Bacteriocin Active Domain 2016-10-10T11:00:00Z 2016-10-11T03:26:24Z We used the Chimera molecular modelling software to determine the sequence of the cytotoxic domain of Colicin iA, we then designed a truncated Colicin Ia, without this domain. The sequence was then codon optimised for E. coli K-12 and synthesised by IDT as a gBlock gene fragment. Using primers we amplified the truncated Colicin Ia with a multiple cloning site containing the following restriction sites: BamHI, KpnI, SalI, BglII and NheI located at the 3' end. The presence of this multiple cloning site will allow for different toxic domains to be fused at the C-terminal end of the truncated colicin and thereby generating synthetic colicins. Colicin Ia is a bacteriocin secreted by E. coli. Colicins typically have 3 domains, a translocation domain, a receptor binding domain, and a cytotoxic domain. If you are looking for a full-length Colicin Ia BioBrick it is located here <partinfo>BBa_K1962000</partinfo>. This part encodes a truncated version of Colicin Ia that lacks the C-terminal bacteriocin domain. The part has an engineered multiple cloning site at the 3' end, preceding the RFC[10] suffix and regulation double stop codons, containing the following in-frame restriction sites: BamHI, KpnI, SalI, BglII and NheI. The presence of this multiple cloning site will allow for different toxic domains to be fused at the C-terminus of the truncated colicin and thereby generating a synthetic colicin in a rapid and facile manner. false false _2429_ 8083 8083 9 false REC[10] compliant false Frank Sargent annotation2491853 1 MCS range2491853 1 1351 1380 annotation2491852 1 Truncated Colicin Ia range2491852 1 1 1350 BBa_K1962002_sequence 1 atgatgtcagacccagtccgcattacgaatcccggcgctgagagcttaggatatgattcggatgggcacgagattatggcagtggacatttatgttaaccccccgcgcgtggacgtttttcacggcacccccccagcttggagcagttttggcaataagacaatctggggtgggaacgaatgggtagacgactcgccaacacgctccgatattgagaaacgtgataaagagattacagcatataagaatacgttatctgcccagcagaaggagaacgagaacaaacgcactgaagcaggcaaacgcctgagtgctgcgatcgcggcacgtgaaaaagacgagaatacgttaaaaactttgcgcgccgggaatgccgatgcggctgacattacgcgtcaagagtttcgtttacttcaagccgaattacgtgaatacggttttcgcacggagatcgcaggatatgatgcattacgcctgcacacagagtcacgcatgttgttcgcggatgctgactcgttacgcatttctccacgtgaagcgcgtagcttaatcgagcaagcggagaagcgtcaaaaagacgctcagaacgctgataagaaagccgcggatatgctggccgaatatgagcgccgtaagggcatccttgacactcgtctttctgaattagaaaaaaacggaggagcagcattagctgttcttgatgctcaacaagctcgtctgttaggacaacagacacgtaacgaccgtgccatctcggaggcgcgtaacaagttgagctcagtaactgagtcgttgaatactgctcgcaacgctttgacccgtgccgagcagcagttgacgcagcaaaagaacactcctgacgggaaaactatcgtttctccggaaaagtttccagggcgcagctcgactaatcactccattgtagtctccggggacccgcgtttcgccgggacgatcaaaatcacaacgagtgccgtcatcgacaaccgtgcaaatctgaactatttactttcccactctggccttgattacaagcgcaacattttaaatgaccgtaaccccgttgttacagaagatgtagaaggtgataagaaaatttacaatgccgaggttgcggagtgggacaagttgcgtcaacgcctgttggatgcacgtaacaaaattacaagcgcagagagtgcagtcaatagtgcccgcaataatttgtccgcacgtacaaatgaacaaaagcacgctaacgatgcattaaacgctctgctgaaggaaaaggaaaacattcgcaaccagctgtcgggcatcaatcaaaagatcgctgaggaaaaacgtaaacaggatgagttaaaggctaccggatccggtaccgtcgacagatctgctagc igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z