Difference between revisions of "Part:BBa K3145002:Design"
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===Design Notes===
 
===Design Notes===
−
There was an illegal XbaI cut site in between our T7 promoter and RBS region in our pY71-sfGFP plasmid that was corrected through site-directed mutagenesis to make our original construct identical to J18912-sfGFP.
+
There was an illegal XbaI cut site in between our T7 promoter and RBS region in our pY71-sfGFP plasmid that was corrected through site-directed mutagenesis to make our T7 promoter and RBS region identical to J18912.
  
 
===Source===
 
===Source===
−
Our original construct (pY71-sfGFP) was readily available in our lab. After site-directed mutagenesis to correct a point mutation in our T7 promoter and RBS region, our original construct was now identical to the J18912-sfGFP construct.
+
Our original construct (pY71-sfGFP) was readily available in our lab. We performed site-directed mutagenesis to remove XbaI cut-site in our T7 promoter and RBS region.
  
 
===References===
 
===References===
 
*<p align="justify"> Kwon, Y.-C. & Jewett, M.C.High-throughput preparation methods of crude extract for robust cell-free protein synthesis. [https://www.nature.com/articles/srep08663|<i>Sci. Rep.</I>] 5,8663; DOI:10.1038/srep08663 (2015).</p>
 
*<p align="justify"> Kwon, Y.-C. & Jewett, M.C.High-throughput preparation methods of crude extract for robust cell-free protein synthesis. [https://www.nature.com/articles/srep08663|<i>Sci. Rep.</I>] 5,8663; DOI:10.1038/srep08663 (2015).</p>

Latest revision as of 22:09, 21 October 2019

J18912-sfGFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 20


Design Notes

There was an illegal XbaI cut site in between our T7 promoter and RBS region in our pY71-sfGFP plasmid that was corrected through site-directed mutagenesis to make our T7 promoter and RBS region identical to J18912.

Source

Our original construct (pY71-sfGFP) was readily available in our lab. We performed site-directed mutagenesis to remove XbaI cut-site in our T7 promoter and RBS region.

References

  • Kwon, Y.-C. & Jewett, M.C.High-throughput preparation methods of crude extract for robust cell-free protein synthesis. Sci. Rep. 5,8663; DOI:10.1038/srep08663 (2015).