Coding

Part:BBa_K942004:Design

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


Shuttle Sequence (SS+B.P.S.)+Der f 2+His


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 573


Design Notes

This composite part has a secretion signal (SS) for an easier purification, maxed out using affinity purification with the C-terminal 6x His tag. As the SS is functional only in yeast, the codon optimization for this sequence was done for a yeast-like codon bias. Also, encrypted in non-translatable sequence after the SS, the T7 promoter (BBa_I712074) and a RBS promote the bacterial expression of the protein. This last feature lets the user transform the same construct for bacterial intracellular expression and extracellular yeast expression.


Source

This part is a mix from the SS of α-mating factor pre-sequence, Der f 2 allergen from Dermatophagoides farinae, and a consensus non-translational yeast mRNA sequence. (Uniprot:Q5TIW0)

References