Part:BBa_K3634002:Design
ATP-Grasp (ATPG) (Codon Optimised for E.coli)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Initially, part BBa_K814001 (Minnesota iGEM, 2012) was taken and the sequence found to include a single EcoR1 illegal restriction site at bp 805. The appropriate synonymous mutation was made in silico to remove this site (a807g). 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 PstI and EcoR1 illegal restriction sites at bp 233 and 787 respectively. The appropriate synonymous mutation was then made in silico to remove these additional site (g231c & a789g). 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_K814001 (Minnesota iGEM, 2012). The part sequence was then optimised for our chosen chassis organism, E.coli, using the IDT codon optimisation tool.
References
Minnesota iGEM 2012 - https://parts.igem.org/Part:BBa_K814001
IDT Codon Optimisation Tool - https://eu.idtdna.com/CodonOpt