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. | + | 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 ( | + | 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=== | ||
− | 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| | + | *<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:
- 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 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).