Device

Part:BBa_K1980012:Design

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


pCopA MymT sfGFP with divergent CueR

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 438
    Illegal NheI site found at 461
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1175
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Codon optimised for E. coli.

The linker between MymT and sfGFP is short and hydrophilic (GlySerGlySerGlySer) to allow the protein domains to fold separately.

CueR is expressed divergent form the pCopA promoter.

Source

The source organism for the MymT protein sequence is Mycobacterium tuberculosis. The CueR and pCopA components of the promoter originate from E. coli.

We ordered this part as codon optimised DNA from IDT.

References

Danya J. Martell, Chandra P. Joshi, Ahmed Gaballa, Ace George Santiago, Tai-Yen Chen, Won Jung, John D. Helmann, and Peng Chen (2015) “Metalloregulator CueR biases RNA polymerase’s kinetic sampling of dead-end or open complex to repress or activate transcription” Proc Natl Acad Sci U S A. 2015 Nov 3; 112(44): 13467–13472

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

Hötzer B., Ivanov R., Altmeier S., Kappl R., Jung G., (2011) "Determination of copper(II) ion concentration by lifetime measurements of green fluorescent protein." Journal of Fluorescence, 21(6), pp. 2143-2153. doi: 10.1007/s10895-011-0916-1