Composite

Part:BBa_K4046610:Design

Designed by: Ashley Jones   Group: iGEM21_Duke   (2021-10-21)


hUBC-Kozak-tdTomato


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 759
    Illegal PstI site found at 1485
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 759
    Illegal PstI site found at 1485
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 759
    Illegal PstI site found at 1485
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 759
    Illegal PstI site found at 1485
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The Kozak sequence is added between the two pieces in order to allow for optimal mammalian expression of the downstream tdTomato gene.


Source

tdTomato is the modified product of a gene originally from mushroom corals in the Discosoma family. The gene was obtained through PCR of a commercially available plasmid pCRISPR-HOT_tdTomato (Addgene, 138567). In normal function, tdTomato is a red fluorescence protein.

hUBC is a constitutive reporter that is associated with the ubiquitin c gene in humans. This gene was obtained through a FUGW-GFP plasmid, generously donated by the Gersbach lab at Duke University. In normal function, the hUBC promoter allows for high levels of expression of associated gene products.

References