Generator

Part:BBa_K942012:Design

Designed by: Luis Mario Leal Garza   Group: iGEM12_Tec-Monterrey   (2012-09-26)

proU+RiAFP+Term


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 951
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The RiAFP is expressed when around 300 mM NaCl added. This part was designed by adding a 6xHis tag for affinity purification to the anti freeze protein from Raghium inquisitor. This part was codon optimized for its expression in E.coli.

Source

The Raghium inquisitor Anti Freeze Protein (RiAFP) is a 137 AA protein that prevents the growth of ice crystals by adsorption to their surface. The proU promoter is from E.coli and works activating transcription when the bacterium is osmotically stressed. (Kristiansen E, 2011)

References

Kristiansen, E. et all (2011) Structural characteristics of a novel antifreeze protein from the longhorn beetle Rhagium inquisitor. Insect Biochem Mol Biol. 41(2):109-17.