BBa_K1378009 1 Binding INPNC-Linker (rTEV)-MVN & GFP 2014-10-05T11:00:00Z 2015-05-08T01:10:12Z This is a composite part so we constructed it using various different smaller parts. INPNC comes from the existing part BBa_K523013. The sequence of MVN can be found in the genomic sequence of Microcystis aeruginosa PCC7806 and the gene was synthesized by a company (sequence can be found in the part BBa_K1378003). GFP is common and comes from the existing part BBa_E0040. The amino acid sequence of the linker can be easily found in papers. This plasmid is an additional testing plasmid for our E. coli- M. aeruginosa binding assay. It differs with the part BBa_K1378007 in that we added a special linker between INPNC and MVN. We inserted two transcription units into one plasmid and they are expressed under the promoters J23105 and J23117 respectively. After this plasmid is transformed into E. coli, the mannan binding protein MVN (Microvirin) will be displayed on the cell surface by INPNC (Ice Nucleation Protein N and C terminal) and the E. coli cell will express GFP as well. There is a linker between INPNC and MVN that can be specifically digested by rTEV protease so we can add rTEV protease to control the binding activity of our E. coli cells. The cell culture with rTEV protease added can serve as the control group for the binding assay. false false _1755_ 0 20995 9 It's complicated false We optimized the sequence of MVN beforehand and chose two different promoters to avoid possible homologous recombination. To ensure the normal functioning of INPNC and MVN we also added a 10aa flexible protein domain linker in between. false Zhang Zijian & Meng Liuyi annotation2421147 1 GFP range2421147 1 1586 2305 annotation2421153 1 MVN range2421153 1 1049 1378 annotation2421148 1 B0015 range2421148 1 2314 2442 annotation2421152 1 B0015 range2421152 1 1385 1513 annotation2421146 1 B0034 range2421146 1 44 55 annotation2421149 1 Linker3 range2421149 1 986 1018 annotation2421150 1 B0030 range2421150 1 1565 1579 annotation2421145 1 Linker2 range2421145 1 1019 1048 annotation2421154 1 INPNC range2421154 1 62 985 annotation2396883 1 J23105 range2396883 1 1 35 annotation2421151 1 J23117 range2421151 1 1522 1556 BBa_K1378009_sequence 1 tttacggctagctcagtcctaggtactatgctagctactagagaaagaggagaaatactagatgaccctggataaagcgctggtgctgcgcacttgtgccaataatatggccgaccattgtggcctgatctggcctgcttctggtacagttgaaagccgctattggcagtctacacgtcgtcacgaaaacgggctggttgggctgctgtggggtgccggaacatccgcctttctgagcgttcatgccgatgctcgttggattgtttgtgaggttgctgttgccgacatcattagcctggaagaaccgggtatggtgaaatttcctcgtgctgaagtggtccatgttggtgatcgtattagcgcctcccatttcatttcagctcgtcaggctgatccagcaagcacatcaacaagcacctctaccagtacactgacaccaatgccaaccgctatccctactcctatgccggctgtagcctctgttactctgcctgtggccgaacaagctcgccacgaagtctttgacgttgcctctgtgtctgctgctgctgctcctgttaacactctgcctgttaccacgcctcaaaatctgcaaacggcgacctatggctcaacactgagtggtgataaccattctcgtctgattgccgggtatggctctaacgaaaccgccggaaaccattctgatctgatcggtggtcacgattgtacgctgatggcaggtgatcaatctcgtctgaccgctggcaaaaattccgttctgaccgctggggcacgttctaaactgatcgggagcgaaggctctactctgtccgccggagaagattctaccctgatctttcgtctgtgggatggtaaacgttatcgtcagctggtcgctcgtacaggtgaaaatggtgtggaagccgacattccgtattatgtgaatgaggacgatgatattgtggacaaaccggacgaagatgatgattggatcgaagtgaaaggtggttctggtgaaaacctttatttccaaggtggtgaaaatttgtattttcaatctggtggtatgccgaacttctctcacacctgctcttctatcaactacgacccgaactctaccatcctgtctgctgaatgccaggctcgtgacggtgaatggatgccgaccgaactgcgtctgtctgaccacatcggtaacatcgacggtgaactccagttcggtgaccagaacttccaggaaacctgccaggactgccacctggagttcggtgacggtgaccagtctgtttggctggtttgcacctgccagaccatggacggtgaatggaaaccgacccagatcctgctggactctcagatcgacaacaacgactctcagctggaaatcggttagtagactagaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagttgacagctagctcagtcctagggattgtgctagctactagagattaaagaggagaaatactagatgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 James Alastair McLaughlin Chris J. Myers 2017-03-06T15:00:00.000Z