Composite
Part:BBa_K2957042:Design
Designed by: Ethan Levy Group: iGEM19_MIT (2019-10-21)
CMV-2xGal4 (Gal4 repressible promoter), IL-8 (CXCL-8), NeonGreen Tag
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 1684
Illegal SpeI site found at 92
Illegal PstI site found at 1127 - 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 92
Illegal PstI site found at 1127 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 901
Illegal BamHI site found at 1143 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 1684
Illegal SpeI site found at 92
Illegal PstI site found at 1127 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 1684
Illegal SpeI site found at 92
Illegal PstI site found at 1127
Illegal AgeI site found at 1690 - 1000COMPATIBLE 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.