Composite

Part:BBa_K2443027

Designed by: Lethbridge iGEM 2017   Group: iGEM17_Lethbridge   (2017-10-18)
Revision as of 00:03, 1 November 2017 by Harlando (Talk | contribs)


EF-Tu optimized for expression in E. coli

EF-Tu (elongation factor thermo unstable) is a prokaryotic elongation factor which catalyzes the binding of an aminoacyl-tRNA (aa-tRNA) to the ribosome. It is a G-protein, and is involved in the selection and binding of an aa-tRNA to the A-site of the ribosome. As a result of its crucial role in translation, EF-Tu is one of the most abundant and highly conserved proteins in prokaryotes. Codon optimized for use in Escherichia coli and contains an N-terminal hexahistidine tag. Under the regulation of T7 Promoter, RBS and double terminator.

Improved Part

Original part:BBa_K1906004 submitted by XJTLU-China 2016

Rational behind improvements: BBa_K1906004 encodes exclusively for the coding region of EF-Tu. In order to improve it and incorporate it into our Next vivo system we have codon optimized for use in E. coli and attached a N-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized EF-Tu to be overexpressed in BL21-Gold DE3 cells by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015).


Sequence and Features


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
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//collections/cell-free_tx-tl
Parameters
None