Difference between revisions of "Part:BBa K2916007:Design"

Revision as of 10:55, 24 September 2019

PheRS 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 BglII site found at 2424
Illegal BamHI site found at 31
Illegal BamHI site found at 1772
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1162
Illegal AgeI site found at 1155
Illegal AgeI site found at 2077
Illegal AgeI site found at 2929
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 2875
Illegal SapI.rc site found at 198
Illegal SapI.rc site found at 2851

Design Notes

The sequence was extracted from the pQE30-His-PheRS plasmid and expressed in the M15 E.coli strain.

Codon optimization : 810 from G to C

Codon optimization : 2151 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/124116/

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.

@@ Line 10: / Line 10: @@
 The sequence was extracted from the pQE30-His-PheRS plasmid and expressed in the M15 E.coli strain.
-Codon optimization : 810 from C to G
+Codon optimization : 810 from G to C
+Codon optimization : 2151 from G to C
 Second stop codon was added in compliance with iGEM standards.