M. tuberculosis strain H37Rv, M. smegmatis strain mc 2 155, and their derivatives were used in this study. All the bacterial strains used in this study are listed in Table . The plasmids used in this study are listed in Table . The targeting sequences are listed in Table . The E. coli strains were cultured in Luria-Bertani broth (LB). M. tuberculosis and M. smegmatis were grown at 37 °C in Middlebrook 7H9 broth or 7H10 plates supplemented with 0.2% glycerol, 0.05% Tween80, 1× albumin dextrose catalase (ADC) (M. smegmatis) or oleic acid ADC (M. tuberculosis) and the appropriate antibiotics. When noted, antibiotics or chemicals were used at the following concentrations: kanamycin (20 μg/mL), leucine (50 μg/mL), Isoniazid (INH) (16 μg/mL) and anhydrotetracycline (ATc) (20 ng/mL).