Description
The pROX expression vector series are for expressing proteins in E. coli cell-free coupled transcription/translation systems, including PURESYSTEM® and The RYTS Kit. Each vector in the series contains optimally positioned upstream T7 promoter and RBS and downstream T7 terminator sequences. In addition, they encode N terminal ProX (see below for explanation) and C terminal hexahistidine tags. Expression templates generated using RYTS Linear Template Kits contain convenient restriction enzyme sites to facilitate subcloning into pROX series expression vectors.
Used in conjunction with Protein Express's CloverDirect™ line of tRNA reagents for site-directed protein functionalization, pROX series expression vectors can be used to express proteins containing unnatural amino acids. CloverDirect™ UAG or CGGG aminoacyl tRNAs are available charged with different unnatural amino acids that confer a range of functionality, including fluorescent-labeling, biotin-labeling, PEGylation, chemical crosslinking and photo-regulation. Targeting incorporation of specific unnatural amino acids is achieved by designing protein expression templates with UAG and/or CGGG codons at user-defined positions. The ProX tag is a special N terminal 14 residue peptide designed for the incorporation of certain large unnatural amino acids that require precise N terminal positioning (codon 9) and a specific codon context (flanked by AAC and TCT). Codon 9 of the ProX tag can be varied between UAG, CGGG or TTT (as a control).
