Difference between revisions of "Part:BBa K2957002:Design"

 
 
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===Design Notes===
 
===Design Notes===
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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. See more on the MIT iGEM 2019 wiki: <a href=https://2019.igem.org/Team:MIT>.
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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 disfunctional. See more on the MIT iGEM 2019 wiki: <a href=https://2019.igem.org/Team:MIT>.
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Latest revision as of 18:52, 22 September 2019


IL-8 (CXCL-8) with NeonGreen tag


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 460
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 460
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 234
    Illegal BamHI site found at 476
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 460
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 460
  • 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 disfunctional. 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