Reporter

Part:BBa_K1179003:Design

Designed by: Nelson Hall   Group: iGEM13_MIT   (2013-09-15)


(Acyl-TyA-mkate-His) A membrane targeting mkate construct with an appended His tag


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 598
    Illegal PstI site found at 847
    Illegal PstI site found at 869
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 598
    Illegal PstI site found at 847
    Illegal PstI site found at 869
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 698
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 598
    Illegal PstI site found at 847
    Illegal PstI site found at 869
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 598
    Illegal PstI site found at 847
    Illegal PstI site found at 869
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1354


Design Notes

Keep in mind that the Acyl-TyA sequence exports the protein into exosomes so protein extraction and other quantification protocols are needed to test successful experimental function. The red fluorescent protein won't be seen in the cells due to the exportation mechanism.

Source

The TyA protein is derived from a yeast retrotransposon virus-like particle and when added with the Acylation tag it acts as an exosome export protein, allowing the mKate to be exported when fused to the TyA.

References