Coding

Part:BBa_K2201205

Designed by: Daniel Bergen   Group: iGEM17_Bielefeld-CeBiTec   (2017-11-01)
Revision as of 16:07, 1 November 2017 by Dbergen195 (Talk | contribs)

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Cysteinyl-lysinyl-tRNA Synthetase

This cysteinyl-lysinyl-tRNA synthetase is based on the work of Nguyen et al. (2011). It is originated from the pyrrolysyl-tRNA synthetase of Methanosarcina barkeri containing a C313V mutation.
Nε-L-cysteinyl-L-lysine (CL) is an amino acid containing a free 1,2-aminothiol group at its side chain. 1,2-aminothiols are an important part of the synthesis of D-Luciferin. D-Luciferin is the substrate of the luciferase of the firefly Photinus pyralis. The synthesis of D-Luciferin is based on a condensation reaction between CL and a cyanobenzothiazole derivative resulting in a covalent bond between two thiazole residues. We designed and synthesized the novel amino acid Nγ‑2‑cyanobenzothiazol‑6‑yl‑L‑asparagine which contains 6-amino-2-cyanobenzothiazole at its side chain. Additionally, we showed that both amino acids are able to undergo the mentioned condensation reaction enabling a system for highly specific binding between peptides and enzymes.
So this synthetase is an important part of our fusing tool. Together with the CLtRNA (K2201206) we developed a aminoacyl-tRNA synthetase/tRNA pair (K2201208) to incorporate CL.

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 492
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None