This study aimed to characterize Escherichia coli (E. coli) isolates from urinary tract infections (UTIs) in Iraq and compare them with international strains. Twenty midstream urine samples from UTI patients were cultured on MacConkey and nutrient agar plates. All bacterial isolates were identified using Vitek 2 system, followed by DNA extraction with the Goya Gene kit. Specific primers were used to detect E. coli, and positive samples were sequenced using the Sanger method. Phylogenetic analysis was conducted to elucidate the genetic relationships among the isolates. The results revealed a high similarity between local Iraqi E. coli isolates and national strains, with minor differences. The phylogenetic tree analysis identified three distinct strains, with an 81% similarity among most isolates, though one international strain was notably distant. The study highlights on rising the frequency of UTIs and the monetary problems they impose on both the governments and patients, emphasizing the need for active drugs that can be rapidly implemented in medical settings. This research underscores the importance of understanding local bacterial populations to inform treatment strategies, as UTIs commonly result from bacterial resistance to existing treatments. UPEC frequently targets the urethra, bladder, and kidneys, initiating infections through colonization and subsequent invasion of the bladder epithelium.