Difference between revisions of "Part:BBa K2443013"
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<h1> Improved Part </h1> | <h1> Improved Part </h1> | ||
<p><b>Original part:</b>BBa_K567015 submitted by SJTU-BioX-Shanghai 2011</p> | <p><b>Original part:</b>BBa_K567015 submitted by SJTU-BioX-Shanghai 2011</p> | ||
| − | <p><b> Rational behind improvements:</b> BBa_K567015 encodes for a truncated MetRS without the anticodon recognition domain. We have improved this part by including the entire coding sequence, which has been codon optimized for optimal expression in E. coli. For easy purification we have included a C- terminal hexahistidine tag with a serine glycine linker. We have also optimized MetRS to be overexpressed in BL21 DE3 gold cell by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015). The original part is also incompatible with biobrick standards. To address this we have removed all illegal cut sites from the construct. </p> | + | <p><b> Rational behind improvements:</b> BBa_K567015 encodes for a truncated MetRS without the anticodon recognition domain. We have improved this part by including the entire coding sequence, which has been codon optimized for optimal expression in E. coli. For easy purification we have included a C- terminal hexahistidine tag with a serine glycine linker. We have also optimized MetRS to be overexpressed in BL21-Gold (DE3) gold cell by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015). The original part is also incompatible with biobrick standards. To address this we have removed all illegal cut sites from the construct. </p> |
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Revision as of 01:06, 31 October 2017
MetRS optimized for expression in E. coli
Methionine tRNA synthetase responsible for attaching methionine onto its tRNA to form an aminoacyl-tRNA. Codon optimized for use in Escherichia coli. Contains a C-terminal histidine tag with a serine glycine linker. Under the regulation of T7 Promoter, RBS and double terminator.
Improved Part
Original part:BBa_K567015 submitted by SJTU-BioX-Shanghai 2011
Rational behind improvements: BBa_K567015 encodes for a truncated MetRS without the anticodon recognition domain. We have improved this part by including the entire coding sequence, which has been codon optimized for optimal expression in E. coli. For easy purification we have included a C- terminal hexahistidine tag with a serine glycine linker. We have also optimized MetRS to be overexpressed in BL21-Gold (DE3) gold cell by putting it under the control of a T7 promoter (BBa_I719005), RBS (BBa_B0034) and double terminator (BBa_B0015). The original part is also incompatible with biobrick standards. To address this we have removed all illegal cut sites from the construct.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]