Extended Data Fig. 3: ΦAP1.1 lysogenization into the S. pyogenes NCTC13743 type II-A CRISPR locus.
(a) S. pyogenes NCTC13743 type II-A CRISPR-cas locus, wild-type and the mutant lacking spacer-repeat units 1 through 10, [Δ1-10]. The ‘t’ indicates three degenerate repeats that contain mutations from the repeat consensus sequence. (b) Sequences of the S. pyogenes NCTC13743 spc2, spc6 and spc9 crRNAs annealed to their targets in ΦAP1.1. The PAM nucleotides are boxed. (c) We verified that the type II-A CRISPR system present in NCTC13743 is capable of restricting a plasmid containing the ΦAP1.1 target sties. Transformation efficiency of the pC194 plasmid or a modified version harboring the three targets for spc2, spc6 and spc9, pTgt2-6-9 from ΦAP1.1, after electroporation of wild-type or [Δ1-10] S. pyogenes NCTC13743 competent cells. Mean + STD of 3 biological replicates are reported. These results demonstrate that the type II-A CRISPR system present in this strain is capable of restricting a plasmid containing the three target sequences present in ΦAP1.1 (d) Diagrams of the type II-A locus of 25 ΦAP1.1 lysogens in S. pyogenes NCTC13743, reconstructed after sequencing of their attL and attR sites.
