Extended Data Figure 10. Inducible curing of a target plasmid.

a, Schematic diagram of plasmids utilized in the plasmid curing experiment. The pGG3 CRISPR plasmid harbors a single spacer (“spc1”) targeting a sequence (“prtspc1”) inserted downstream of the P_xyl/tet* inducible promoter in pWJ153. b, Agarose gel electrophoresis of linearized plasmid DNA purified from both anhydrotetracycline-treated (+ATc) and untreated (−ATc) cultures at the indicated timepoints. 10 kb, 5 kb, and 4 kb size markers are indicated. Picture is representative of a single technical replicate. c, Colony forming units (cfu) recovered from cultures analyzed in panel (b) at each time point. Cells were plated with selection for either Cm^R cfu (green) or Cm^R, Erm^R cfu (blue). Targeting of the pWJ153 plasmid via induction with ATc (filled circles) is accompanied by a severe drop in erythromycin-resistant cfu relative to untreated cultures (open circles). Error bars: mean ± s.d. (n=3).