Difference between revisions of "Part:BBa K2443028"
| Line 4: | Line 4: | ||
EF-Ts is a prokaryotic elongation factors. EF-Ts acts at the guanine nucleotide exchange factor for EF-Tu, catalyzing the release of guanosine diphosphate from EF-Tu. This enables EF-Tu to bind to a new guanosine triphosphate molecule, release EF-Ts, and go on to catalyze another aminoacyl tRNA addition. EF-Ts has been Codon optimized for use in <i>Escherichia coli</I> and contains a C-terminal tag. Under the regulation of T7 Promoter, RBS and double terminator. | EF-Ts is a prokaryotic elongation factors. EF-Ts acts at the guanine nucleotide exchange factor for EF-Tu, catalyzing the release of guanosine diphosphate from EF-Tu. This enables EF-Tu to bind to a new guanosine triphosphate molecule, release EF-Ts, and go on to catalyze another aminoacyl tRNA addition. EF-Ts has been Codon optimized for use in <i>Escherichia coli</I> and contains a C-terminal tag. Under the regulation of T7 Promoter, RBS and double terminator. | ||
| + | |||
| + | <h1> Improved Part </h1> | ||
| + | <p><b>Original part:</b>BBa_K1906000 submitted by XJTLU-China 2016</p> | ||
| + | <p><b> Rational behind improvements:</b> BBa_K1906000 encodes exclusively for the coding region of EF-Ts. 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 C-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized EF-Ts 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). | ||
Revision as of 01:39, 31 October 2017
EF-Ts optimized for expression in E. coli
EF-Ts is a prokaryotic elongation factors. EF-Ts acts at the guanine nucleotide exchange factor for EF-Tu, catalyzing the release of guanosine diphosphate from EF-Tu. This enables EF-Tu to bind to a new guanosine triphosphate molecule, release EF-Ts, and go on to catalyze another aminoacyl tRNA addition. EF-Ts has been Codon optimized for use in Escherichia coli and contains a C-terminal tag. Under the regulation of T7 Promoter, RBS and double terminator.
Improved Part
Original part:BBa_K1906000 submitted by XJTLU-China 2016
Rational behind improvements: BBa_K1906000 encodes exclusively for the coding region of EF-Ts. 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 C-terminal hexahistidine tag with a serine glycine linker for easy purification. We have also optimized EF-Ts 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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 65
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]