BBa_K1978000 1 BBa_K1978000 TorA-BtuF 2016-10-12T11:00:00Z 2016-10-17T02:45:40Z All DNA sequences were synthesized by IDT on the basis of the amino acid sequence of TorA in E.coli and BtuF from E. coli (K12). The TorA-BtuF Biobrick consists of a TorA signal linked to BtuF, which is a vitamin B12 binding protein. The TorA signal sequence allows export of fully-folded proteins through the inner membrane via the TAT(Twin-Arginin)export system. This enables export of vitamin B12 out of the cell. The TorA sequence codes for a peptide that harbours a twin-arginine motif. This is vital for the recognition by the Tat system. Moreover, an AxA motif is present, which leads to cleavage by the leader peptidase I (Palmer & Berks 2012). MNNNDLFQASRRRFLAQLGGLTVAGMLGPSLLTPRRATA BtuF is the periplasmic binding protein relevant for uptake of vitamin B12 through the outer membrane that is associated with the ABC transporter BtuCD (Kandt et al., 2006). BtuF is the periplasmic binding protein. It has a size of 30.19 kDa and is composed of two globular domains, between which vitamin B12 is bound, linked by a rigid interdomain α-helix (Karpowich et al., 2003). The TorA signal sequence and the sequence for BtuF are connected by a long linker. false false _2445_ 29934 29978 9 true The sequences were designed by codon optimization and exclusion of restriction sites present in iGEM prefix and suffix. The linker needed to be constructed in a way that is similar to the BtuF amino acid sequence. false Larissa Kr??ger annotation2497197 1 Linker range2497197 1 118 132 annotation2497199 1 BtuF range2497199 1 133 927 annotation2497194 1 TorA Signal Peptide range2497194 1 1 117 BBa_K1978000_sequence 1 atgaacaataacgatctctttcaggcatcacgtcggcgttttctggcacaactcggcggcttaaccgtcgccgggatgctggggccgtcattgttaacgccgcgacgtgcgactgcggcgcaagcggcgactgctaagtccctgtttcgcgcacttgttgcattatcctttttggccccgctttggctgaatgcagctccccgggtcattacactttcccctgcaaacaccgaattggcatttgctgctggcattacaccagtcggagtatcctcgtactccgattaccccccccaagctcaaaaaatcgaacaagtgtcgacgtggcagggaatgaatctggagcggatcgtagcattgaaaccggatcttgtgatcgcgtggagaggcgggaatgcggagcgccaggtggaccagttggcctcccttggtattaaggtaatgtgggtcgatgcaacttccatcgagcagatagcgaatgcgctgagacaacttgccccttggagtccccagcccgacaaagcggagcaagctgcccaatccttgctggatcagtacgcccaattaaaggctcaatatgctgataaaccgaagaagagagtgttcctgcaatttggcatcaatccgccgtttacctcaggtaaggagtcgatccagaaccaggtcttggaggtttgcggaggagaaaatatcttcaaagactctcgtgtaccgtggccgcaggtttcacgcgagcaagttctggcacgctcacctcaggctatcgtcataacagggggaccagaccaaattcctaaaataaagcaatactggggagagcagctgaaaattccagttatccctttaacatcggactggtttgaaagagcgtcaccacgcattatccttgcagcccaacaattgtgtaatgcgctttcccaagtggac 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