Difference between revisions of "Part:BBa K3634000:Design"

Revision as of 15:20, 20 July 2020

3-Dehydroquinate Synthase (DHQS) (Codon Optimised for E.coli)

Design Notes

Initially, part BBa_K814000 (Minnesota iGEM, 2012) was taken and the sequence found to include a single EcoR1 illegal restriction site at bp 111. The appropriate synonymous mutation was made in silico to remove this site (g111a). This allowed for the part to be used at RFC[10] and RFC[1000] standard without codon optimisation. The sequence was then optimised using the IDT codon optimiser tool. This resulted in a new SapI illegal restriction site at bp 650. The appropriate synonymous mutation was then made in silico to remove this additional site (a657g). This allowed for the part to be used at RFC[10] and RFC[1000] standard with codon optimisation.

Source

The initial genomic sequence came from Anabaena variabilis ATCC 29413 which can be obtained from part BBa_K814000 (Minnesota iGEM, 2012). The part sequence was then optimised for our chosen chassis organism, E.coli, using the IDT codon optimisation tool (https://eu.idtdna.com/CodonOpt).

References

Minnesota iGEM 2020 - https://parts.igem.org/Part:BBa_K814000

IDT Codon Optimiser Tool - https://eu.idtdna.com/CodonOpt