Coding
LysRS

Part:BBa_K2916006:Design

Designed by: Konstantinos Ragios   Group: iGEM19_EPFL   (2019-07-24)
Revision as of 13:19, 23 September 2019 by Konstantinos (Talk | contribs) (→‎Design Notes)


LysRS protein equipped with a 6x HIS affinity tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1121
    Illegal XhoI site found at 1516
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE 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 : 1344 from T to A

Codon optimization : 480 from C to G

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.