Composite

Part:BBa_K2916027:Design

Designed by: Konstantinos Ragios   Group: iGEM19_EPFL   (2019-09-25)


Expression of PheRS in E.coli


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2533
    Illegal BamHI site found at 140
    Illegal BamHI site found at 1881
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1271
    Illegal AgeI site found at 1264
    Illegal AgeI site found at 2186
    Illegal AgeI site found at 3038
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2984
    Illegal SapI.rc site found at 307
    Illegal SapI.rc site found at 2960


Design Notes

The protein was expressed in pQE30 in M15 E.coli strain.


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.