A gram-negative, rod-shaped, non-motile, gamma and UV-tolerant bacterium, Hymenobacter sp. DG25A, was isolated from a soil sample collected in South Korea. The isolated strain demonstrated high level of resistance against gamma irradiation, with a D10 value of 6 kGy. The complete genome of Hymenobacter sp. DG25A was found to consist of a single chromosome (3,777,136 bp). The bacterium was isolated from a gamma ray-irradiated soil sample and was found to contain enzymes involved in the accumulation of manganese, which protects proteins from oxidation by reactive oxygen species. The genome also encodes enzymes for the nucleotide excision repair pathway, which leads to the efficient removal of double-strand breaks caused by ionizing radiation such as gamma and UV rays. An understanding of these genomic features can potentially be used in several biotechnological applications such as for the degradation of harmful contaminants or radioactive waste.