Composite

Part:BBa_K2916014:Design

Designed by: Konstantinos Ragios   Group: iGEM19_EPFL   (2019-07-25)


Expression of AlaRS 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 573
    Illegal BglII site found at 780
    Illegal BamHI site found at 746
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1184
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2584
    Illegal BsaI site found at 2761
    Illegal BsaI.rc site found at 1133
    Illegal SapI site found at 1868


Design Notes

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

Source

Sebastian Maerkl Lab at EPFL, Switzerland.

http://www.addgene.org/124103/

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.