A yellow-pigmented bacterial strain, GS03T, was isolated from sediment in a branch of the Nackdong River in Sangju, Korea. Cells were observed to be Gram-negative, aerobic and rod-shaped with gliding motility, and to be positive for catalase and oxidase. Growth was found to occur at 4-30°C (optimum 25°C), at pH 7.0-8.5 (optimum pH 7.5) and at NaCl 0% (optimum NaCl 0%, w/v). The major cellular fatty acids (> 10% of the total) were identified as iso C15:0, iso C15:1 G, C15:1ω6c, iso C15: 0 3-OH and iso C17: 0 3-OH. The major respiratory quinone was found to be menaquinone MK-6. The genome sequence of GS03T is 3.1Mb with G+C content of 36.1mol%. The major polar lipids of the isolate were identified as phosphatidylethanolamine, three unidentified aminolipids, two unidentified phospholipids, an unidentified lipid and an unidentified aminophospholipid. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain GS03T clusters with Flavobacterium paronense KNUS1TT, with similarity of 96.8%. The phenotypic, phylogenetic, and chemotaxonomic characteristics indicate that strain GS03T represents a novel species of the genus Flavobacterium, for which the name Flavobacterium sangjuense sp. nov. is proposed. The type strain is GS03T (= FBCC 502459T = KCTC 62568T = JCM 32764T).