Composite

Part:BBa_K2957022:Design

Designed by: Ye Cheng Zheng   Group: iGEM19_MIT   (2019-10-19)


TRE, IL-8 (CXCL-8) with NeonGreen tag (C-terminal)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1399
    Illegal PstI site found at 850
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 850
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 624
    Illegal BamHI site found at 866
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1399
    Illegal PstI site found at 850
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1399
    Illegal PstI site found at 850
    Illegal AgeI site found at 1405
  • 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: 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