Figure 1.

Maps and the Cloning Sites of the pCasPA/pACRISPR System

(A) Map of the pCasPA plasmid. The expressions of both the Cas9 protein and the λ-Red system were driven by the L-arabinose-inducible araB promoter (P_araB). The counter-selectable marker sacB was used for plasmid curing after editing. tetA, the tetracycline-resistance marker in E. coli and P. aeruginosa; oriV, the origin of replication; trfA, the essential gene for initiation of plasmid replication.

(B) Map of the pACRISPR plasmid. trc Promoter, the sgRNA expression promoter; bla, the carbenicillin-resistance marker in E. coli and P. aeruginosa; mSF, a broad-host-range origin; ColE1, a replication origin for E. coli; BsaI sites, Golden Gate assembly of spacers; XbaI and XhoI sites, Gibson assembly of repair arms; sacB, the counter-selectable marker for plasmid curing after editing.

(C) Sequence of the cloning sites of the pACRISPR plasmid.