Composite

Part:BBa_K2443013

Designed by: Lethbridge iGEM 2017   Group: iGEM17_Lethbridge   (2017-10-18)


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 hexahistidine 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) 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


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