Type strain is Mycobacterium aquicola MiE-22 (GCA_965234515.1), isolated from 5 m depth from Lake Milada, Czechia (date: 2019-10-15), via high-throughput dilution to extinction cultivation. MiE-22 has a genome size of 3.4 Mbp with a genomic GC content of 66.3%, contains 3 rRNA genes and 57 tRNAs. The genome is a high-quality draft consisting of 3 contigs. The genome contains genes encoding rhodopsins. No genes for flagella or pilus assembly and chemotaxis were annotated. Pathways for taurine degradation, assimilatory sulfate reduction and the biosynthesis of all amino acids except for histidine and aspartate were predicted. Further, pathways for many vitamins and cofactors (riboflavin, pyridoxal, NAD, coenzyme A, biotin, tetrahydrofolate, heme, cobalamine and menaquinone) were identified. The closest cultivated relative is Mycobacterium massilipolynesiensis M26 (GCF_001494595.1), with an average amino acid identity of 71.31% and average nucleotide identity of 76.47%. Current GTDB classification (R220): d__Bacteria; p__Actinomycetota; c__Actinomycetia; o__Mycobacteriales; f__Mycobacteriaceae; g__Mycobacterium; s__Mycobacterium sp945897705.