Composite

Part:BBa_K2957007:Design

Designed by: Ethan Levy   Group: iGEM19_MIT   (2019-10-19)


hEF1a, IL-8 (CXCL-8), NeonGreen Tag


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1256
    Illegal PstI site found at 699
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 699
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 473
    Illegal BamHI site found at 715
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1256
    Illegal PstI site found at 699
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1256
    Illegal PstI site found at 699
    Illegal AgeI site found at 81
    Illegal AgeI site found at 1262
  • 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

Parts of this part were ordered from IDT/Twist and then assembled with other parts (from iGEM/parts in our parts collection/from the Weiss lab) into this plasmid.

References