Difference between revisions of "Part:BBa K2916007:Design"
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The sequence was extracted from the pQE30-His-PheRS plasmid and expressed in the M15 E.coli strain. | The sequence was extracted from the pQE30-His-PheRS plasmid and expressed in the M15 E.coli strain. | ||
| − | Codon optimization : | + | Codon optimization : 810 from G to C |
| − | Second stop codon was added in compliance with iGEM standards. | + | Codon optimization : 2151 from G to C |
| + | |||
| + | Second stop codon was added in compliance with iGEM standards. | ||
===Source=== | ===Source=== | ||
| Line 21: | Line 23: | ||
===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 02:02, 22 October 2019
PheRS 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 2424
Illegal BamHI site found at 31
Illegal BamHI site found at 1772 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1162
Illegal AgeI site found at 1155
Illegal AgeI site found at 2077
Illegal AgeI site found at 2929 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2875
Illegal SapI.rc site found at 198
Illegal SapI.rc site found at 2851
Design Notes
The sequence was extracted from the pQE30-His-PheRS plasmid and expressed in the M15 E.coli strain.
Codon optimization : 810 from G to C
Codon optimization : 2151 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/124116/
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.