PyMOL (Schrödinger) was used to visualize the PAM contacts of a previously published structure55 (PDB 5f9r), and amino acid or nucleobase substitutions were introduced with the mutagenesis feature. For each amino acid substitution, we manually evaluated all possible sidechain rotamers to identify plausible configurations that positioned polar contacts between 2.8 and 3.5 Å apart without producing steric clashes. The R1332K substitution yielded various plausible rotamers, and we arbitrarily chose one for visualization in all figures.

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