Coding

Part:BBa_K2957013:Design

Designed by: Ethan Levy   Group: iGEM19_MIT   (2019-09-22)


CCL5 with NeonGreen tag (C-terminal)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 442
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 442
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 458
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 442
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 442
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We used Modular Cloning Golden Gate Type II Assembly, so we had to consider how parts came together. We also used the natural secretion tag given in the part so we could use it to test chemokine secretion success by HEKs. We also had to consider which terminal the fluorescence/protein tag was added so as not to render the chemokine dysfunctional. See more on the MIT iGEM 2019 wiki: <a href=https://2019.igem.org/Team:MIT>.


Source

This part was ordered from IDT/Twist Biosciences as a gBlock and then assembled into a plasmid. The sequence was taken from this site: <a href: https://www.uniprot.org/uniprot/P10145> and then codon-optimized using IDT's codon optimization tool. The NeonGreen tag was a part in the Weiss lab database.

References