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Cas9 Nuclease NLS, S. pyogenes (NLS Cas9), is an RNA-guided endonuclease that catalyzes site specific cleavage of double stranded DNA. The location of the break is within the target sequence 3 bases from the NGG PAM (Protospacer Adjacent Motif). The PAM sequence, NGG, must follow the targeted region on the opposite strand of the DNA with respect to the region complementary sgRNA sequence. Cas9 Nuclease NLS, S. pyogenes contains a single Simian virus 40 (SV40) T antigen nuclear localization sequence (NLS) on the C terminus of the protein.