Supplemental materials and methods; HPLC and LC-HRMS analysis of the metabolites produced from CXR14::pCHW201 (Fig. S1); 1H NMR and 13C NMR data for POL-B (Fig. S2), POL-J (Fig. S3), and POL-C (Fig. S5); in-frame deletion of polG on pCHW201 and HPLC/LC-HRMS analysis of the target metabolites produced by the CXR14::pCHW201/ΔpolG recombinant (Fig. S4); 1H NMR, 13C NMR, and 2D-NMR data for thymine POL-C (Fig. S6); LC-HRMS analysis of CPOAA produced from the CXR14::pCHW201/ΔpolG recombinant (Fig. S7); bioinformatics analysis of PolG with its homologs (Fig. S8); LC-HRMS analysis of the PolG-catalyzed reactions (Fig. S9); HPLC and LC-HRMS analysis of PolG-catalyzed reaction using NIK-Cx as the substrate (Fig. S10); structural comparison of RizA and PolG proteins (Fig. S11); SDS-PAGE analysis and in vitro characterization of PolG variants (Fig. S12); LC-HRMS analysis of the proposed mechanism of PolG-catalyzed reaction (Fig. S13); phylogenetic analysis of PolG against other ATP-dependent ligases (Fig. S14); optimized sequence for PolG (Table S1); PCR primers used in this study (Table S2).
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