BBa_K1185005 1 Switch E. coli L-form Switch 2013-10-25T11:00:00Z 2015-05-08T01:09:37Z All the sequences come from the Escherichia coli K12 genomic sequence. The ftsI section in the BioBrick was taken from end of the ftsI gene 1268-1767bp (500bp). Sequence acquired through EcoCyc (EG10341). The murE section was taken from the start of the murE gene 1-500bp. Sequence from EcoCyc (EG10621). The Pspac promoter were taken from the registry designed by the 2007 Cambridge iGEM team (BBa_1746666). Ampicilin resistance marker gene was taken from the expression vector pSTAR (gb|AF047654.1|AF047654). This is and engineered BioBrick derived from the BBa_K1185000 (L-form switch for ''Bacillus subtilis'')which can be used as a switch to enable the model Gram-negative bacteria ''Escherichia coli'' to switch between cell-walled rod form and wall-less L-form. The BioBrick contains homologous regions to the ftsI (homolog to pbpb in Bacillus subtilis)and murE genes to allow integration into the chromosome of E. coli via homologous recombination. Within the BioBrick there is a IPTG inducible promoter Pspac. This promoter is upstream of the murE coding sequence when integrated into the chromosome of E. coli, placing murE operon expression under the control of IPTG. The murE operon is responsible for the synthesis of enzymes involved in production of peptidoglycan synthesis. Disruption of this pathway can be utilised to down-regulate production of the cell wall. After integration of the BioBrick, the expression of murE operon is controlled by the presence, or absence, of IPTG(via the Pspacpromoter). When IPTG is present, murE operon genes are expressed and functional cell wall is produced. When IPTG is not present the cell wall is no longer produced and cells can transition to the cell wall-less L-form phenotype. The BioBrick also contains a cat gene, conferring Ampicilin resistance. This can be used to select for transformants in rod form. This BioBrick only functions in enabling switching between walled cells and wall-deficient L-forms in E. coli. We are giving opportunities for future iGEM team to work with cell wall-less E. coli. This BioBrick should work in a similar manner as the BBa_K1185000 (B. subtilis Switch BioBrick). The characterization of this BioBrick is something that future iGEM can do. false false _1498_ 0 16794 9 Not in stock false The BioBrick does not contain any illegal restriction sites. All of the sequences have been taken directly from E. coli genome so there is no need for codon optimization. false Vincent Leonardo annotation2370730 1 murE Homologous region range2370730 1 1894 2393 annotation2370728 1 BBa_1746665 range2370728 1 1819 1876 annotation2370726 1 ftsI Homologous region range2370726 1 1 500 annotation2370729 1 murE endogenous range2370729 1 1877 1893 annotation2370727 1 ampR range2370727 1 501 1818 BBa_K1185005_sequence 1 taacaccattacagttagcgcgagtctacgcaactatcggcagctacggcatttatcgcccactgtcgattaccaaagttgaccccccggttcccggtgaacgtgtcttcccggaatccattgtccgcactgtggtgcatatgatggaaagcgtggcgctaccaggcggcggcggcgtgaaggcggcgattaaaggctatcgtatcgccattaaaaccggtaccgcgaaaaaggtcgggccggacggtcgctacatcaataaatatattgcttataccgcaggcgttgcgcctgcgagtcagccgcgcttcgcgctggttgttgttatcaacgatccgcaggcgggtaaatactacggcggcgccgtttccgcgccggtctttggtgccatcatgggcggcgtattgcgtaccatgaacatcgagccggatgcgctgacaacgggcgataaaaatgaatttgtgattaatcaaggcgaggggacaggtggcagatcgtaagacatcgatgacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataataatggtttcttagacgtcaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcgcccttattcccttttttgcggcattttgccttcctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgagtgggttacatcgaactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcacttttaaagttctgctatgtggcgcggtattatcccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgacttggttgagtactcaccagtcacagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgagtgataacactgcggccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgtaactcgccttgatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagcaatggcaacaacgttgcgcaaactattaactggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggctcgagtaagttgactttatctacaaggtgtggcataatgtgtgtaattgtgagcggataacaattaataaggggtgaggctggcgtggcagatcgtaatttgcgcgaccttcttgctccgtgggtgccagacgcaccttcgcgagcactgcgagagatgacactcgacagccgtgtggctgcggcgggcgatctctttgtagctgtagtaggtcatcaggcggacgggcgtcgatatatcccgcaggcgatagcgcaaggtgtggctgccattattgcagaggcgaaagatgaggcgaccgatggtgaaatccgtgaaatgcacggcgtaccggtcatctatctcagccagctcaacgagcgtttatctgcactggcgggccgcttttaccatgaaccctctgacaatttacgtctcgtgggcgtaacgggcaccaacggcaaaaccacgactacccagctgttggcgcagtggagccaactgcttggcgaaatcagcgcggtaatgggcaccgttggtaacggcctgctggggaaagtgatcccgacagaaaatacaaccggttcggcagtcgatgttcagca 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