Background: Emerging antibiotic resistance and Extended-Spectrum Beta-Lactamase (ESBL) producing Escherichia coli causing different nosocomial infections are rapidly increasing at alarming levels and it poses a major health burden in the 21st century globally. Objective: The aim of this study is to determine the distribution of blaTEM gene ESBLproducing E. coli from clinical specimens in different hospitals in Erbil city. Methods: A total of 200 samples were collected from (sputum, urine, wound) from patients attended public hospitals in Erbil city. The isolation and identification of Escherichia coliand antimicrobial susceptibilitywere performed by using Vitek 2 compact -lactamase production in E.coli was confirmed by using both Double disk diffusion and Standard disk diffusion techniques. Moreover, PCR technique was used for genotypic detection of an ESBL gene blaTEMaccording to the standard protocol. Results: Out of 200 samples 60(30%) of E. coli isolates and the highest rate of E. coli isolates were obtained from the urine samples (20%) and the lowest number was isolated from sputum specimens (2.5%). The ESBL-producing Escherichia coli isolates were detected using double disk synergy test (76.7%) in comparison to standard disk diffusion test (80%). Genotypic screening results confirmed that all ESBL-producing E. coli isolates (66.7%) were carried blaTEM gene (700 bp) in clinical specimens (50% urine, 13.33% wound and 3.33% sputum). All ESBL-positive E. coli isolates showed high rates of susceptibility to Carbapenems antibiotic group including Imipenem (83.3%), Meropenem (81.7%), and Ertapenem (80.0%). Conclusions: -lactamase gene in ESBL-producing E. coli observed in this study for the first time is considered as alarming because there is a limited treatment options remained for infections. Attempts to reduce the dissemination of multi-drug resistant E. coli through compliance with strict hospital infection control and prevention standards are imperative. Findings of this study may help clinicians selecting appropriate antimicrobial therapy in patients with different infections caused by ESBLproducing E. coli.