Reporter

Part:BBa_K1980009:Design

Designed by: Sam Garforth   Group: iGEM16_Oxford   (2016-10-11)


pCusC CusR RFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 527
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Frustrating point mutation (T to C) atnulceotide 706 converting a Val to Ala residue in the CusR DNA binding domain. The reading frame is not affected.


Codon optimised to E. coli. A BspH1 restriction site was included at the start codon of the sfGFP so that other proteins with a compatible start (such as the copper chelators we ligated into our pBAD vector) could be ligated into this location

Source

CusR and pCusC originate from the E. coli genome. Ordered as codon optimised DNA from IDT.

References

Yamamoto K, Ishihama A. (2005) “Transcriptional response of Escherichia coli to external copper.” Mol Microbiol. 2005 Apr;56(1):215-27.

Sambandam Ravikumar, Van Dung Pham, Seung Hwan Lee, Ik-keun Yoo, Soon Ho Hong (2012) “Modification of CusSR bacterial two-component systems by the introduction of an inducible positive feedback loop” Journal of Industrial Microbiology & Biotechnology June 2012, Volume 39, Issue 6, pp 861–868