Difference between revisions of "Part:BBa K2916004:Design"
Line 8: Line 8:
 
===Design Notes===
 
===Design Notes===
  
−
The sequence was extracted from the pET21a-CysRS-His plasmid and expressed in the BL21 (DE3) E.coli strain.
+
The sequence was extracted from the pET21a-IleRS-His plasmid and expressed in the BL21 (DE3) E.coli strain.
 +
 
 +
Codon optimization : 1179 from G to C
 +
 
 +
Second stop codon was added in compliance with iGEM standards.  
  
 
===Source===
 
===Source===
  
−
http://www.addgene.org/124107/
+
Sebastian Maerkl Lab at EPFL, Switzerland.
 +
 
 +
http://www.addgene.org/124112/
  
 
===References===
 
===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
 
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.

Revision as of 11:46, 20 September 2019


IleRS protein equipped with a 6x HIS affinity tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 2780
    Illegal NotI site found at 2842
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 843
    Illegal BamHI site found at 1346
    Illegal XhoI site found at 2851
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1003
    Illegal AgeI site found at 1921
    Illegal AgeI site found at 2380
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1465


Design Notes

The sequence was extracted from the pET21a-IleRS-His plasmid and expressed in the BL21 (DE3) E.coli strain.

Codon optimization : 1179 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/124112/

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.