Abstract
We present the complete genome sequence of Sphingomonas sp. gentR, a strain exhibiting high-level resistance to gentamicin (MIC = 40 mg/mL). The genome was assembled from hybrid Illumina and Nanopore sequencing data into a gap-free sequence of 4.0 Mbp, comprising one chromosome and two plasmids. A total of 3,692 coding sequences were predicted, with comprehensive functional annotation revealing genes associated with antibiotic resistance, stress adaptation, and metabolic diversity. Three confirmed resistance genes-ANT(2″)-Ia, ANT(3″)-IIa, and Sul1-were co-localized within a genomic island on plasmid B. This dataset provides insight into the genetic basis of high-level aminoglycoside resistance in Sphingomonas and serves as a valuable resource for studying horizontal gene transfer, environmental adaptation, and bioremediation potential. The genome sequence is publicly available under GenBank accessions CP144670-CP144672 and China National Genomics Data Center (accession number GWHDOHA00000000).