The complete mito-genomes of three species (O. lacepedii, O. rebecca and Odontamblyopus sp.) were sequenced, their genomic structure examined, and their genome organization, arrangement and codon usage analyzed. Phylogenetic Bayesian and ML analyses were conducted, using a concatenated set of 12 protein-coding genes, and adding 16 other species of gobies (Gobiidae). The mitogenome sequences of O. lacepedii, O. rebecca and Odontamblyopus sp. were all circular double-strand molecules, 17245 bp, 17009 bp and 17004 bp long, respectively. Compared with other bony fishes, the three species shared the similar features in gene arrangements, base composition and tRNA structure. The control region spanned 1571 bp, 1336 bp and 1332 bp in O. lacepedii, O. rebecca and Odontamblyopus sp., respectively, and was A+T-rich. The length variation of control region was due to the tandemly repeated sequences. Three species were only detected termination-associated sequence domain (TAS) and conserved sequence blocks domain (CSB-1, CSB-2 and CSB-3). The Bayesian and ML tree topologies of 19 Gobiidae represented two groups: one large group consisted of Amblyopinae, Gobionellinae, Oxudercinae and Sicydiinae, and the other was the monophyletic Gobiinae. Phylogenetic analysis also demonstrated clade Amblyopinae included all three species and that O. lacepedii has been proved to be a much closer affinity to Odontamblyopus sp. than O. rebecca. Our study theoretically provided a supplementary proof and significant information for future taxon studies.