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Inle Lake is an ancient lake in Myanmar, which is an important area with unique and diverse fauna. Its ichthyofauna is believed to have formed non‐radiatively, but the historical processes are poorly understood. To elucidate the mechanisms that shape species diversity in this moderately isolated biogeographical 'island', this study focused on a typical endemic fish of Inle Lake, Channa harcourtbutleri (Channidae, Anabantiformes), with its widely distributed sister species, C. limbata, and estimated the historical distribution and diversification processes of the endemic fish based on genome‐wide polymorphism (MIG‐seq) and mitochondrial DNA data. Channa harcourtbutleri contained two genetically and morphologically distinct groups inhabiting Inle Lake and the surrounding rivers respectively. These two groups were genetically the closest to each other; however, the riverine group showed some similarity to the closely related species, C. limbata from Southeast Asia. The mtDNA haplotypes of the endemic species were not monophyletic; most of the riverine group had haplotypes identical or close to those of C. limbata from the upper Irrawaddy and Salween rivers. The time tree suggested that C. harcourtbutleri diverged from C. limbata in the early Pleistocene and then experienced secondary contact with C. limbata in the late Pleistocene. Genetic and morphological differentiation within C. harcourtbutleri suggests that local adaptation to different environments has played an important role for the coexistence of its two forms with some reproductive isolation. Further, the results highlight the importance of multiple colonization and allopatric speciation in shaping biodiversity in the long‐term, moderately isolated environments. [ABSTRACT FROM AUTHOR]