their respective pET28b multicloning regions. The cloned promoter DNA was engineered with
these restriction enzyme sites, and was constructed from two synthetic oligonucleotides that
were annealed and digested accordingly (Table 2). This strategy directly added the native
E. coli promoter, displacing the existing T7 promoter region. The newly constructed vectors
were named pCRC01UVRA, pCRC02UVRA, and pCRC03UVRA, according to the notation
introduced in the above paragraph. Verification of expression was performed by SDS-PAGE
analysis of the protein from EPI300 E. coli containing pCRC03UVRA as well as
phenotypic assays.