Difference between revisions of "Part:BBa K2683018"

 
 
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<partinfo>BBa_K2683018 short</partinfo>
 
<partinfo>BBa_K2683018 short</partinfo>
  
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CysRs mow in the pSb1C3 biobrick standard
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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.
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https://static.igem.org/mediawiki/2018/f/f2/T--Lethbridge--CysOE.PNG
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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.
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https://static.igem.org/mediawiki/2018/c/cf/T--Lethbridge--Cyspur.PNG
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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.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 03:24, 17 October 2018


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.

T--Lethbridge--CysOE.PNG

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.

T--Lethbridge--Cyspur.PNG

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1065
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 931