The research focused on analyzing the UGT gene family in Cannabis sativa, which plays a crucial role in the plant's metabolism and glycosylation of secondary metabolites. The study identifed 125 UGTs using conserved plant secondary product glycosyltransferase (PSPG) motif amino acid sequences. These UGT genes were categorized into 17 groups (A-Q) through phylogenetic analysis, showing their distribution across 10 chromosomes in C. sativa. The expansion of the CsUGT gene family was attributed to tandem and duplication events, as suggested by gene duplication analysis. Furthermore, the study found various cis-acting regulatory elements related to phytohormones and stress responses in CsUGT promoter regions. Subcellular localization analysis revealed that CsUGT is present in the cytoplasm, chloroplast, and nucleus. The study revealed that CsUGT plays a signifcant role in various biological processes, cellular components, and molecular functions as highlighted by Gene Ontology analysis. Additionally, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that some CsUGTs are associated with the biosynthesis of secondary metabolites. This research provides valuable insights into the genomic organization, evolutionary history, and potential regulatory mechanisms of UGT genes in C. sativa. It lays the foundation for further exploration of their specifc biological roles and potential applications in the plant's metabolism and stress responses. These fndings contribute to a better understanding of the UGT gene family and its relevance to the metabolic pathways in C. sativa.