The complete mitochondrial genome of the Greek nine-spined stickleback Pungitius hellenicus was obtained using Illumina high-throughput sequencing of genomic DNA. The genome was 16 713 bp long, and contained 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and a control region.
The arrangement of the genes was identical to that of other Gasterosteidae fishes. However, the control region of P. hellenicus contained three copies of imperfect repeated sequences (72-78 bp in single motifs), while P. pungitius and P. tymensis have one or two copies.
Nucleotide identity between P. hellenicus and three other Pungitius species across all the 37 genic regions was 93.0% to 95.5%.