Reporter

Part:BBa_K1179001:Design

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


(Acyl-TyA-eGFP) A membrane targeting eGFP construct


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1384
    Illegal SpeI site found at 1359
    Illegal PstI site found at 604
    Illegal PstI site found at 853
    Illegal PstI site found at 875
    Illegal PstI site found at 1389
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1384
    Illegal SpeI site found at 1359
    Illegal PstI site found at 604
    Illegal PstI site found at 853
    Illegal PstI site found at 875
    Illegal PstI site found at 1389
    Illegal NotI site found at 1410
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1384
    Illegal BglII site found at 704
    Illegal BamHI site found at 1353
    Illegal XhoI site found at 1417
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1384
    Illegal SpeI site found at 1359
    Illegal PstI site found at 604
    Illegal PstI site found at 853
    Illegal PstI site found at 875
    Illegal PstI site found at 1389
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1384
    Illegal SpeI site found at 1359
    Illegal PstI site found at 604
    Illegal PstI site found at 853
    Illegal PstI site found at 875
    Illegal PstI site found at 1389
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 537
    Illegal BsaI.rc site found at 889
    Illegal BsaI.rc site found at 1342


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. GFP won't be seen in the cells due to this 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 eGFP to be exported when fused to the TyA. The eGFP comes from the jellyfish Aequorea victoria.

References