At present, few morphological descriptions are available for Acentrogobius species and there exist some confused issues on the species classification and phylogeny. In this study, we first determined and described the complete mitochondrial genome of Acentrogobius sp. The complete mitogenome sequence is 17 083 bp in length, containing 13 protein-coding genes, two rRNA genes, 22 tRNA genes, a putative control region (CR), and a light-strand replication origin (OL). The overall base composition is 28.9% A, 26.2% T, 28.5% C, and 16.4% G, with a slight AT bias (55.1%). To furthermore validate the new determined sequences, phylogenetic trees involving all the Gobiidae species available in GenBank database were constructed. These results are expected to provide useful molecular data for species identification and further phylogenetic studies of Gobiiformes.