Part:BBa_K3145002:Design

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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).