Difference between revisions of "Part:BBa K2957001:Design"

 
 
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__NOTOC__
 
__NOTOC__
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<partinfo>BBa_K2957001 short</partinfo>
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<partinfo>BBa_K2957099 short</partinfo>
  
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<partinfo>BBa_K2957001 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K2957099 SequenceAndFeatures</partinfo>
  
  
 
===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 dysfunctional. See more on the MIT iGEM 2019 wiki: <a href=https://2019.igem.org/Team:MIT>.
  
  

Latest revision as of 18:51, 22 September 2019


hEF1a, IL-8 (CXCL-8)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 3667
    Illegal PstI site found at 2430
    Illegal PstI site found at 2935
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2430
    Illegal PstI site found at 2935
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2684
    Illegal BamHI site found at 3290
    Illegal BamHI site found at 3569
    Illegal XhoI site found at 3083
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 3667
    Illegal PstI site found at 2430
    Illegal PstI site found at 2935
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 3667
    Illegal PstI site found at 2430
    Illegal PstI site found at 2935
    Illegal NgoMIV site found at 197
    Illegal NgoMIV site found at 2818
    Illegal AgeI site found at 2025
    Illegal AgeI site found at 2196
    Illegal AgeI site found at 3673
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 2073
    Illegal SapI.rc site found at 9


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

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 Flag tag was a part in the Weiss lab database.

References