Herein we present the cytochrome oxidase subunit I (COI) based phylogeography of the dice snake (Natrix tessellata) in Eastern Europe and The Balkans. Eight haplotypes were identified. Phylogenetic reconstruction has showed 3 major clades: 1) Greece, 2) Balkans, Central and Eastern Europe, 3) Eastern Europe (Ukraine and Russia only). Our data are in concordance with the results of other studies of the species' phylogeography. The northernmost isolated population of the dice snake in Lipetsk province, Russia have no differences from neighboring Ukrainian populations and seems to be the result of recent colonization during the Holocene climatic optimum.