The Ring-necked Pheasant (Phasianus colchicus) is a large, chicken-like bird native to North America and Asia, recognizable by its long, pointed tail, and small head with long neck. An important game bird in North America, it consumes a variety of foods including seeds, grains, grasses, leaves, roots, fruits, nuts, and insects. Recognizing the importance of the Ring-necked Pheasant, a genetic diversity analysis was conducted in Pakistan using mitochondrial DNA cytochrome oxidase 1 (COI) technique. Thirty different samples were collected, and after DNA extraction, quantification, primers optimization, and PCR, the sequences were aligned with reported sequences on NCBI. A phylogenetic tree was constructed using MEGA6 software, and the DNA barcode for the species was identified. The study provided insights into the genetic structure and phylogeny of the species. These unique COI sequences offer a reliable method for the quick identification of Ring-necked Pheasant, aiding in the prevention of illegal poaching and supporting conservation management efforts.