Composite

Part:BBa_K2957034:Design

Designed by: Malik Aaron George   Group: iGEM19_MIT   (2019-10-19)


TRE, C5a-NeonGreen


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4128
    Illegal XbaI site found at 4175
    Illegal SpeI site found at 3240
    Illegal PstI site found at 621
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4128
    Illegal SpeI site found at 3240
    Illegal PstI site found at 621
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4128
    Illegal BglII site found at 2089
    Illegal BamHI site found at 637
    Illegal XhoI site found at 2205
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4128
    Illegal XbaI site found at 4175
    Illegal SpeI site found at 3240
    Illegal PstI site found at 621
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4128
    Illegal XbaI site found at 4175
    Illegal SpeI site found at 3240
    Illegal PstI site found at 621
    Illegal AgeI site found at 4181
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3768
    Illegal SapI.rc site found at 3741


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