Antibiotic resistance in Enterobacteriaceae, particularly Extended Spectrum β-lactam resistance, is increasingly dominated by the activation of constantly expressed genes encoding effective drug modifying enzymes. Forty three Gram-negative bacilli isolates were screened for their Extended Spectrum β-lactamase production. They were isolated from different clinical urine samples and identified by standard biochemical reactions. Antibacterial susceptibility testing including disc diffusion method using 13 antibiotics discs including (ceftriaxone, azithromycin, aztreonam, cefotaxime, ceftazidime, clindamycin, streptomycin, norfloxacillin, chloramphenicol, ciprofloxacin, sulphamethoxazole-trimethoprim, and ampicillin/clavulanic acid) was done for all isolates. The antibiotic susceptibility test, disk diffusion method, and double disc synergy test indicated that seven enteric uropathogenic isolates were ESBL producers during the present study. They recorded diameters of inhibition zones as ≤ 18, ≤ 8, ≤ 19, and ≤ 8 mm against cefotaxime (CTX), ceftazidime (CAZ), aztreonam (ATM), and ceftriaxone (CRO). Genotypically, blaTEM genes are the most common, with (100%) occurrence in all five enteric tested uropathogens, followed by blaSHV and blaCTX genes (60%). Analyzing the 16S rRNA sequence confirmed that the most potent ESBL-producing bacteria (U60) isolate were identified as Escherichia coli U60.1 and with accession numbers MW173246, in GenBank.