Composite
Part:BBa_K3145002:Design
Designed by: Yazmin Camacho Group: iGEM19_US_AFRL_CarrollHS (2019-09-09)
J18912-sfGFP
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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. [https://www.nature.com/articles/srep08663%7C "Sci. Rep.<i/>" 5, 8663; DOI:10.1038/srep08663 (2015).