Difference between revisions of "Part:BBa K2916000:Design"

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===Design Notes===
 
===Design Notes===
  
−
The protein was cloned in the pQE30 vector and expressed in the M15 E.coli strain
+
The sequence was extracted from the pET16b-His-ArgRS plasmid and expressed in the BL21(DE3) E.coli strain
 +
 
 +
Codon optimization : 1512 from A to T
 +
 
 +
Codon optimization : 1566 from G to C
 +
 
 +
Second stop codon was added in compliance with iGEM standards.
  
 
===Source===
 
===Source===
  
−
Escherichia coli gene alaS
+
Sebastian Maerkl Lab at EPFL, Switzerland.
−
SJQSFNOAPZFvbe
+
 
 +
https://www.addgene.org/124104/
  
 
===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
 +
 +
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 01:45, 22 October 2019


ArgRS protein equipped with a 10x HIS affinity tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 76
    Illegal SapI.rc site found at 1495


Design Notes

The sequence was extracted from the pET16b-His-ArgRS plasmid and expressed in the BL21(DE3) E.coli strain

Codon optimization : 1512 from A to T

Codon optimization : 1566 from G to C

Second stop codon was added in compliance with iGEM standards.

Source

Sebastian Maerkl Lab at EPFL, Switzerland.

https://www.addgene.org/124104/

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.