Difference between revisions of "Part:BBa K2916006:Design"
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The sequence was extracted from the pET21a-LysRS-His plasmid and expressed in the BL21 (DE3) E.coli strain. | The sequence was extracted from the pET21a-LysRS-His plasmid and expressed in the BL21 (DE3) E.coli strain. | ||
| − | Codon optimization : | + | Codon optimization : 1245 from A to G |
Codon optimization : 1179 from G to C | Codon optimization : 1179 from G to C | ||
Revision as of 13:17, 23 September 2019
LysRS protein equipped with a 6x HIS affinity tag
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1121
Illegal XhoI site found at 1516 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 475
Design Notes
The sequence was extracted from the pET21a-LysRS-His plasmid and expressed in the BL21 (DE3) E.coli strain.
Codon optimization : 1245 from A to G
Codon optimization : 1179 from G to C
Codon optimization : 1179 from G to C
Second stop codon was added in compliance with iGEM standards.
Source
Sebastian Maerkl Lab at EPFL, Switzerland.
http://www.addgene.org/124114/
References
Cell-free translation reconstituted with purified components. Shimizu Y, Inoue A, Tomari Y, Suzuki T, Yokogawa T, Nishikawa K, Ueda T. Nat Biotechnol. 2001 Aug;19(8):751-5. doi: 10.1038/90802. 10.1038/90802 PubMed 11479568
Lavickova, Barbora, and Sebastian J. Maerkl. A Simple, Robust, and Low-Cost Method to Produce the PURE Cell - Free System. preprint, Synthetic Biology, 18 Sept. 2018. DOI.org (Crossref), doi:10.1101/420570.