Aims: The study aimed to evaluate the type and frequency of integrons in multi-drug-resistant Escherichia coli isolated from clinical and environmental samples in Basrah, Iraq. Methodology and results: The MDR E. coli strains were identified using both conventional and molecular techniques. Antibiotic susceptibility was tested using the Kirby-Bauer method. The results revealed that out of 62 samples studied, 58 (93.54%) of the local E. coli isolates were resistant to at least three different antibiotics. These MDR E. coli strains showed high resistance to penicillin, erythromycin, and tetracycline, with rates of 62 (100%), 57 (91.93%), and 50 (80.64%), respectively. Additionally, we investigated the presence of integrons in MDR Escherichia coli using specific primers through the PCR technique. The results showed that 32 (51.61%) of the total isolates contained IntI1 (491 bp in size), while 5 (8.06%) harbored IntI2 and 6 (9.67%) harbored IntI3 (788 bp and 600 bp, respectively). A significant association (P ≤ 0.05) was also observed between the presence of IntI1 and resistance to several antibiotics in MDR E. coli. Conclusion, significance and impact of the study: We concluded that integrons, particularly IntI1, are major genetic determinants in MDR E. coli strains. These elements play a crucial role in the horizontal transfer of drug resistance genes in clinical isolates of Escherichia coli.