Composite

Part:BBa_M50026:Design

Designed by: Maurice Chiang   Group: Stanford BIOE44 - S11   (2016-10-27)


Gold Detection Device with Promoter RBS and GolS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 485
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

These parts include a promoter that binds a GolS inducer, a ribosome binding site (RBS), and a gene coding for GolS, a protein originating from Salmonella typhimurium that forms a protein-metal complex with gold (Au). GolS, upon binding Au, dimerizes and then binds a recognition site on the promoter, forming the basis for our sensor device. In short, this construct is a gold-sensitive promoter for E. coli.

Our construct (promoter + RBS + GolS) was combined with the E. coli sensor cassette plasmid. This allows us to connect our construct with sequences for ampicillin resistance, origin of replication and GFP expression.

Sequence Changes

While the GolS gene was originally drawn from York iGEM 2013, we made a modification to the start codon. We changed the start codon to -AUG- to increase the efficiency of translation in E. coli. This is because E. coli more frequently uses AUG to initiate translation.

Image: 250 pixels


Source

This part was drawn from York iGEM in 2013, found at this link: https://parts.igem.org/Part:BBa_K1127008 The E. coli sensor cassette was provided to us through Stanford BioE 44.

References