Genetic diversity and genetic relatedness among 50 genotypes from eight countries, including Iran, Afghanistan, Turkmenistan, Syria, Lebanon, India, Yemen, and the United States located in two continents of Asia and the America, were assessed using SCoT markers. A total of 213 bands were produced; 100% of them were polymorphic; the average polymorphism information content (PIC) was 0.39. The mean Nei's gene diversity and Shannon's index were 0.33 and 0.49, respectively. Analysis of molecular variance suggested significant genetic differences within pomegranate populations. 99% of variance occurs within the populations, whereas 1% of the variation was recorded among the populations of pomegranate. Cluster analysis using SCoT markers able to group genotypes based on their geographical origins. Based on cluster analysis, the genotypes studied were divided into two main groups. The first group included most Asian genotypes, while American genotypes along with some Asian genotypes were in the second group. In the first group, Iranian genotypes were grouped with genotypes from Afghanistan and India. In the second group, the genotypes belonging to the America were in the same group as most of the genotypes of Turkmenistan. According to the present study, SCoT markers can be used to evaluate genetic diversity, identification and DNA fingerprinting pomegranate genotypes of different origins. This information can be used in breeding programs and the management of pomegranate collections.