The Fasin Rainbow fish, scientifically known as Melanotaenia fasinensis, is highly sought after by aquarium enthusiasts due to its vibrant colors and adaptability to artificial aquatic environments. This species is endemic to the karst landscape of the Bird’s Head region in Papua, Indonesia, and belongs to the Melanotaeniidae family. Discovered relatively recently in 2010, it has been designated as endangered by the International Union for Conservation of Nature (IUCN) in 2021. However, there is currently insufficient data regarding its phylogenetic positioning. To address this gap, our study employed next-generation sequencing to analyze the entire mitochondrial genome of M. fasinensis. The mitochondrial genome consists of 13 protein-coding genes, 22 transfer RNA genes, and two ribosomal RNA genes, with a total length of 16,731 base pairs. The base composition of the mitogenome revealed percentages of 27.76% adenine (A), 27.34% thymine (T), 16.15% guanine (G), and 28.75% cytosine (C). Our phylogenetic analysis, based on nucleotide sequences mitogenome, indicated that M. fasinensis occupies a relatively basal position within the Melanotaenia genus. This study provides valuable molecular insights for further exploration of phylogeography and evolutionary history, not only for M. fasinensis but also for other members of the Melanotaenia genus.