Difference between revisions of "Part:BBa K3634017:Design"

Latest revision as of 14:50, 8 August 2020

ccaR (Codon Optimised for E.coli)

Design Notes

In the initial sequence obtained from Tabor's pJT122 plasmid, an illegal EcoR1 site is found. Uppsala-Sweden iGEM 2011 removed the site by carrying out point mutagenesis of A4C and A6C to obtain the part BBa_K592002. The part sequence was then subject to codon optimisation using the IDT codon optimisation tool to allow optimum expression of ccaR in the E.coli chassis. No new illegal restriction sites were introduced by this step.

Source

ccaR can be found in the genome of Synechocystis PCC 6803. The sequence can be obtained from BBa_K592002 (Uppsala-Sweden iGEM, 2011 - initially from Tabor's pJT122 plasmid). The part sequence was then fully optimised for our chosen chassis organism, E.coli, using the IDT codon optimisation tool.

References

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

Hirose Y., Shimada T., Narikawa R., Katayama M., Ikeuchi M. 2008. Cyanobacteriochrome CcaS is the green light receptor that induces the expression of phycobilisome linker protein. PNAS. 105(28): p9528-9533. DOI: 10.1073􏰃/pnas.0801826105

Tabor J.J., Levskaya A., Voigt C.A. 2011. Multichromatic control of gene expression in Escherichia coli. J Mol Biol. 405(2): p315–324. DOI: 10.1016/j.jmb.2010.10.038

Uppsala-Sweden iGEM 2011 - https://parts.igem.org/Part:BBa_K592001

Schmidl S.R., Sheth R.U., Wu A., Tabor J.J. 2014. Refactoring and Optimization of Light-Switchable Escherichia coli Two-Component Systems. ACS Synth Biol. 3: p820-831. DOI: 10.1021/sb500273n