Composite

Part:BBa_K3634009:Design

Designed by: Laurence Seeley   Group: iGEM20_St_Andrews   (2020-08-05)
Revision as of 18:46, 5 August 2020 by Ls268 (Talk | contribs) (→‎References)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


ccaS + ho1 + pcyA (Optimised for E.coli)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 2449
    Illegal NheI site found at 2472
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2126
    Illegal BglII site found at 2879
    Illegal BamHI site found at 1465
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1497
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Schmidl et al. (2014) used both promoter and RBS libraries to produce the system termed CcaSR v2. Two plasmids are used for constitutive expression of ccaS and ccaR as this was found to increase output of sfgfp, measured quantitatively. Promoter and RBS combinations were also selected to maximise PCB production.

Source

The genomic sequences for all three CDS are IDT codon optimised for E.coli, taken from the native Synechocystis sp. PCC6803. Design of the composite part can be fully attributed to the work of Schmidl S.R., Sheth R.U., Wu A. and Tabor J.J. in the paper 'Refactoring and Optimization of Light-Switchable Escherichia coli Two-Component Systems'. Individual sequences for the parts can be found at BBa_K3634006, BBa_K3634007 and BBa_K3634008.

References

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

Schmidl SR., Sheth RU., Wu A., Tabor JJ. 2014. Refactoring and Optimisation of Light-Switchable Eschericia coli Two-Component Systems. ACS Synth. Biol. 3: p820-831. DOI: 10.1021/sb500273n

Ong NT., Tabor JJ. 2018. A Miniaturized Escherichia coli Green Light Sensor with High Dynamic Range. ChemBioChem. 19: p1255-1258. DOI: 10.1002/cbic.201800007

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