Bioinformatics and whole-genome verification data for: CYP123, a sterol C4-methyl oxidase, acts with CYP51 to initiate mycobacterial lanosterol catabolism

This record contains the supporting data for Yuan et al. (2026), "CYP123, a sterol C4-methyl oxidase, acts with CYP51 to initiate mycobacterial lanosterol catabolism", in two parts. The comparative-genomics half contains sequence similarity networks of the cytochrome P450 family PF00067, for the family Mycobacteriaceae and for all bacteria, with their cluster assignments, genome neighbourhood data, and the sequence sets behind the alignment figures. These were produced with the Enzyme Function Initiative web tools in July and August 2025 against UniProt release 2025-03 and ENA data downloaded in June 2025. They establish that CYP123 and CYP51 lie in a genomic neighbourhood conserved across Mycobacteriaceae and in the wider bacteria, together with a short-chain dehydrogenase, a ferredoxin, an aldehyde dehydrogenase and a second cytochrome P450. The strain-verification half contains Nanopore whole-genome sequencing, assembly, annotation and edit verification for the eight Mycolicibacterium smegmatis strains used in the study: the laboratory wild type, two CRISPR frameshift alleles of cyp123, two ORBIT replacements of cyp123, one ORBIT replacement of cyp51, and two complemented strains. It includes the analysis pipeline with pinned environments and every derived table behind the verification report. Reads and assemblies are also archived at the European Nucleotide Archive under study accession PRJEB128689. Start with README.pdf for the record as a whole, and CYP123_WGS_verification_report.pdf for the strain verification. Because Zenodo presents a record as a flat list, the directory structure is held inside two archives: CYP123_bioinformatics_data.zip for the comparative-genomics data, and CYP123_WGS_zenodo_package_2026-10-26.tar.gz for the whole-genome material. The two Cytoscape sessions are supplied uncompressed, outside the archives, because they are what most readers will want. Every other file on the record is covered by CHECKSUMS.sha256. Crystal structures are in the Protein Data Bank under 9TJO and 9TJP and are not duplicated here.