We isolated floating goby Gymnogobius petschiliensis mitochondrial DNA by long-polymerase chain reaction (Long-PCR) with conserved primers, and sequenced the mitogenome by primer walking using flanking sequences. The G. petschiliensis mitochondrial DNA has 16,424 bp and its structural organization is similar to the mitochondrial DNAs of other fish, and mammals. We analyzed phylogenetic relationships derived from the mitochondrial cytochrome b gene. We report the basic characteristics of the G. petschiliensis mitochondrial genome including its structural organization and the base composition of the rRNAs, tRNAs and protein-coding genes as well as characteristics of tRNAs. These features are used to analyze phylogenetic relationship among the 60 species of the genus Gymnogobius.