Difference between revisions of "Part:BBa K3145002:Design"

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===References===
 
===References===
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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).
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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).]

Revision as of 03:53, 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.

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.

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).