The strain was engineered using lambda red recombination, employing the counterselection strategy by Bubnov et al., (2020). Briefly, helper plasmid pRedCmOcr containing lamdba red genes, restriction protection gene ocr, and PL promoter controlled cmr gene was used to transform the starting strain MG1655 from NRRL. A clhokneo cassette that contains neomycin resistance gene, hok gene and cl repressor gene that represses PL promoter with 110 bp HA was made. This was integrated between the lac operator and lacZ  via electroporation after prior lambda red induction using rhamnose. Successful integration of the cassette was screened via the presence of kanamycin resistance and chloramphenicol sensitivity. These colonies were then verified via colony PCR of the integration locus and comparison with the starting strain. The verified clhokneo integrated strain was then further engineered to replace the clhokneo cassette with the T7 RNA polymerase.  This was done by electroporating the c1hokneo strain with template T7 RNA polymerase containing 110 bp HAs. Strains that had successful integration of the T7 RNA polymerase was screened via Chloramphenicol resistance and Kanamycin sensitivity. They were further verified via colony PCR, gDNA PCR of the integration locus and DNA sequencing. IPTG induction in this strain was also verified by successful expression of recombinant mCherry fusions using pTi plasmid with an expression cassette under lac promoter and operator.