Difference between revisions of "Part:BBa K2916003:Design"
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__NOTOC__
 
__NOTOC__
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<partinfo>BBa_K2916001 short</partinfo>
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<partinfo>BBa_K2916003 short</partinfo>
  
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<partinfo>BBa_K2916001 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K2916003 SequenceAndFeatures</partinfo>
  
  
 
===Design Notes===
 
===Design Notes===
−
The sequence was extracted from the pET16b-His-ArgRS plasmid and expressed in the BL21 (DE3) competent E.coli strain  
+
The sequence was extracted from the pET21a-GlyRS-His plasmid and expressed in the BL21 (DE3) competent E.coli strain  
 +
 
 +
Codon optimization : 1509 from A to G
  
 
===Source===
 
===Source===
  
−
http://www.addgene.org/124104/
+
Sebastian Maerkl Lab at EPFL, Switzerland.
 +
 
 +
https://www.addgene.org/124110/
  
 
===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 10:53, 20 September 2019


GlyRS 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 XhoI site found at 2989
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1737
    Illegal AgeI site found at 471
    Illegal AgeI site found at 2859
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 958


Design Notes

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

Codon optimization : 1509 from A to G

Source

Sebastian Maerkl Lab at EPFL, Switzerland.

https://www.addgene.org/124110/

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.