Tyrosyl synthetase, library plasmid from Methanococcus jannashii
Tyrosyl synthetase from wild type Methanococcus jannashii in pSB1C3, designed as library plasmid for the randomerization of certain positions and following selection cycles for the incorporation of non canonical amino acids or unnatural bases. Tyrosyl tRNA synthetase from Methanococcus jannashii in pSB1C3, designed as library plasmid for the randomerization of certain positions and following selection cycles for the incorporation of non-canonical amino acids or unnatural bases. In addition BBa_K2201400, the part BBa_K2201411 contains an mRFP (BBa_J04450) in the position of the synthetases binding pocket.
Usage and Biology
We designed and cloned this part to generate a tRNA/synthetase based on the Methanococcus jannashii wild type tyrosyl-tRNA/aminoacyl synthetase, which is able to incorporate 2-Nitro-L-phenylalanine, used for the photocleaving of the polypeptide backbone. Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC
We cloned a library of more than 130,000 clones, including more than 27,672 different TyrRS variants out of 32,768 possible sequence variants, analyzed with MiSeq, Illumina next generation sequencing. These sequences code for more than 8,787 different peptides.
We used this TyrRS library as a basis to select an tRNA/aminoacyl synthetase pair, able to incorporate 2-Nitrop-L-phenylalanine. Therefore we used our selection system, consisting of a positive (BBa_K2201900) and negative selection plasmid (BBa_K2201901) .