Part:BBa_K2683018
Cysteine tRNA synthetase (CysRS) in pSB1C3
Cysteine tRNA synthetase is a protein that has the function of attaching cysteine onto its tRNA to form an aminoacyl-tRNA [1]. The part is Codon optimized for use in Escherichia coli, contains a C-terminal hexahistidine tag with a serine glycine linker, has T7 Promoter, medium RBS and double terminator. These original designs were done by the 2017 Lethbridge iGEM team (http://2017.igem.org/Team:Lethbridge) This part was improved by the 2018 iGEM team by placing this composite part into the pSB1C3 standard.
Figure 1- The overexpression of CysRS run on a 12% SDS-PAGE gel. Samples were taken at the point of induction and in 1 hour increments over time. Overexpression can be seen at 53.5kDa.
Figure 2- Nickel affinity chromatography purification of CysRS protein. Samples were run on a 12% SDS-PAGE. Purified samples are in elutions 1-5 at about 53kDa.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1065
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 931
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