BBa_K1962006 1 BBa_K1962006 Truncated Colicin E9 Lacking Bacteriocin Active Domain 2016-10-10T11:00:00Z 2016-10-11T07:02:58Z We used the Chimera molecular modelling software to determine the position of the DNase domain of Colicin E9, we then designed a truncated Colicin E9, without this domain. The protein sequence was then back translated into DNA and codon optimised for E. coli K-12 before being synthesised by IDT as a gBlock gene fragment. Using oligonucleotide primers we then amplified the truncated colicin E9 with a multiple cloning site containing the following restriction sites: BamHI, KpnI, SalI, BglII and NheI at the 3' end. Colicins are anti-bacterial proteins produced by some strains of E. coli that typically have three domains: a translocation domain; a receptor binding domain; and a cytotoxic domain. This biobrick encodes a truncated version of Colicin E9 which lacks the C-terminal bacterocin domain, which is this case is a DNase. 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, or other polypeptides, to be fused at the C-terminus of the truncated colicin and thereby generating a synthetic colicin or novel fusion protein in a rapid and facile manner. false false _2429_ 8083 8083 9 false Compliant with RFC[10]. false Frank Sargent annotation2491957 1 Truncated E9 range2491957 1 1 1356 annotation2491958 1 MCS range2491958 1 1357 1386 BBa_K1962006_sequence 1 atgtctggcggcgacggacgcggccataacactggagcgcatagcacttccgggaacatcaatggggggcccactgggattggcgtctctggcggggcgagtgacggcagtgggtggagttcagaaaacaatccttggggcggcggatcgggatcggggattcattgggggggcggttctggtcgcggtaatggtggaggaaacggtaactccggcggtgggagtggcacagggggtaacttgtcggcagttgcggctcccgtggcgtttggttttcctgctttatcgacaccaggagcaggcgggttggcagttagtatcagcgcctcggaactttctgccgcgattgcaggcattatcgctaagctgaaaaaggttaatttaaagtttacgccgtttggagtggtattatccagcttgatccccagtgagatcgcaaaggatgacccgaatatgatgagcaagatcgtcacttcccttccggccgacgatatcacggaaagccctgtttctagtcttcccttggataaggccaccgtgaatgtcaatgttcgtgttgtagatgacgtcaaagacgaacgccaaaacattagtgtagtcagcggagtccctatgagcgtgcctgttgtagacgccaagcccaccgagcgtccgggtgtctttaccgcgtctattcccggagcacctgtgcttaacattagtgtgaacgactcgaccccagccgtccaaacactttctccgggcgtgacaaataacaccgataaagatgttcgtccagctggttttacccagggcggtaatacccgtgatgctgttattcgttttcccaaagattccggtcacaacgccgtttacgtctcggtgtcagatgtcctttccccagatcaagtcaagcaacgtcaagatgaggaaaaccgccgccaacaagaatgggatgctacacacccggtggaggcagcagaacgtaattacgaacgtgcacgcgccgagttaaaccaggccaatgaggacgtggcgcgcaaccaagaacgccaagccaaggcggtacaggtctacaactcgcgtaagagtgagcttgatgcagcaaacaaaacacttgcagacgccattgcggaaattaaacagtttaatcgcttcgctcatgatccaatggctggtggccatcgtatgtggcagatggctggattgaaagcgcagcgcgctcagactgacgtaaataacaagcaagcggcctttgatgctgccgcgaaagaaaaaagtgacgccgacgcagctctgtcagcggcacaagagcgccgcaagcagaaggagaacaaagaaaaagatgccaaggataaacttgataaggagagtaaaggatccggtaccgtcgacagatctgctagc 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