BBa_K1978001 1 BBa_K1978001 TorA-GlmS 2016-10-12T11:00:00Z 2016-10-13T04:56:34Z All DNA sequences were synthesized by IDT on the basis of the amino acid sequence of TorA in E.coli and GlmS from Clostridium cochlearium. The TorA-GlmS Biobrick consists of a TorA signal linked to GlmS, 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 GlmS is the B12-binding subunit of glutamate mutase (Glm) from Clostridium cochlearium. The mechanism by which the enzyme uses adenosylcobalamin is highly similar to methylmalonyl coenzyme A mutase. Glm catalyzes the reversible rearrangement of (2S)-glutamate to (2S,3S)-3-methylaspartate (Leutbecher et al., 1992). The assembly of the active enzyme, an α2β2 tetramer, is mediated by coenzyme B12. While GlmS as the smaller subunit (14.8 kDa) binds B12, the larger subunit GlmE harbors the substrate binding site (Zelder et al., 1994). false false _2445_ 29978 29978 9 false 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 GlmS amino acid sequence. false Larissa Kr??ger annotation2497226 1 Linker range2497226 1 118 132 annotation2497225 1 TorA Signal Peptide range2497225 1 1 117 annotation2497227 1 GlmS range2497227 1 133 540 BBa_K1978001_sequence 1 atgaacaataacgatctctttcaggcatcacgtcggcgttttctggcacaactcggcggcttaaccgtcgccgggatgctggggccgtcattgttaacgccgcgacgtgcgactgcggcgcaagcggcgactgagaagaaaactatcgtcctgggcgttattgggtctgattgccatgccgtcggaaataaaatactggatcatgcttttaccaacgccgggttcaatgtcgtcaatattggagttttaagccctcaagaggttttcattaaagccgccatagaaacgaaggcggacgccattctgctttcatcactttacggccagggcgagatcgactgcaagggcctgcgtcaaaaatgtgatgaggctgggttggaaggaatcctgttatacgttgggggtaacatcgttgtcgggaagcaacactggcctgacgtcgaaaaaagattcaaggacatgggttacgatagagtctacgcccctgggaccccgccagaagtaggaattgctgatttaaaaaaagacttaaacattgag 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