Part:BBa_K2201205

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 mutationю

N^ε-L-cysteinyl-L-lysine 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 N^γ‑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]