Difference between revisions of "Part:BBa K4579008:Design"

Latest revision as of 10:15, 12 October 2023

CvaC15 - Signal peptide

Assembly Compatibility:

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-<partinfo>BBa_K4579008 SequenceAndFeatures</partinfo>
-Because CvaC15 must end in a double glycine residue in order to be recognized and cleaved by CvaB [2], we could not use the Type 3a and 3b convention for the overhangs of our signal peptide and microcin parts. To ensure that the overhangs allow the double glycine to be maintained immediately upstream of the main microcin coding sequence, we positioned the BsaI restriction site that would be added at the 3’ end of CvaC15 such that the last 4 nucleotides of the sequence (i.e., the last nucleotide in the first glycine codon and the entirety of the second glycine codon) became the suffix for the part. We then simply used the reverse complement of those nucleotides as the prefix that would be added to the 5’ end of any microcin that was used in our microcin expression plasmid so that, when the two parts were assembled, the microcin sequence essentially “completes” the signal peptide sequence. Using NEBridge GetSet, we confirmed that this overhang did not have a significant likelihood of mis-ligating with the overhangs of our other parts.
+__NOTOC__
+<partinfo>BBa_K4579008 short</partinfo>
+<partinfo>BBa_K4579008 SequenceAndFeatures</partinfo>