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The RYTS (Remarkable Yield Translation System) is a cell-free E. coli extract-based protein synthesis system that includes all essential components for coupled transcription/translation reactions. The E. coli extract is prepared according to a unique method developed by RIKEN for highly efficient protein production. For template generation we recommend expression plasmids from Protein Express (pROX-FL92.1: ProX-tag + His-tag) or BioComber (pURE1-4: no tags).
In-gel detection of TAMRA-labeled proteins produced by the RYTS Kit. 2UAG: UAG codon inserted after initiator AUG codon. ProX tag: ProX tag* fused to the N-terminus. 25ul of RYTS rxn was on an SDS-PAGE gel. White arrows indicate TAMRA-labeled proteins. *ProX tag is an N-terminal peptide designed for large non-natural amino acids whose efficient incorporation requires precise N-terminal positioning (codon 9) and a specific codon context (flanked by AAC and TCT). Originally developed for the 4 base CGGG codon, the ProX tag can also function to promote N-terminal incorporation of unnatural amino acids using the amber codon.