Difference between revisions of "Part:BBa K2916004:Design"
Konstantinos (Talk | contribs) |
Konstantinos (Talk | contribs) |
||
| Line 21: | Line 21: | ||
===References=== | ===References=== | ||
| + | |||
| + | <i> | ||
| + | McClain, William H. “Rules That Govern TRNA Identity in Protein Synthesis.” Journal of Molecular Biology, vol. 234, no. 2, Nov. 1993, pp. 257–80. DOI.org (Crossref), doi:10.1006/jmbi.1993.1582. | ||
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 | 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. | + | 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.</i> |
Latest revision as of 01:54, 22 October 2019
IleRS protein equipped with a 6x HIS affinity tag
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 2780
Illegal NotI site found at 2842 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 843
Illegal BamHI site found at 1346
Illegal XhoI site found at 2851 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1003
Illegal AgeI site found at 1921
Illegal AgeI site found at 2380 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1465
Design Notes
The sequence was extracted from the pET21a-IleRS-His plasmid and expressed in the BL21 (DE3) E.coli strain.
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/124112/
References
McClain, William H. “Rules That Govern TRNA Identity in Protein Synthesis.” Journal of Molecular Biology, vol. 234, no. 2, Nov. 1993, pp. 257–80. DOI.org (Crossref), doi:10.1006/jmbi.1993.1582.
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.