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The global spread of antimicrobial resistance genes (ARGs) is a major public health concern. Mobile genetic elements (MGEs) are the main drivers of this spread by horizontal gene transfer (HGT). <i>Escherichia coli</i> is widespread in various environments and serves as an indicator for monitoring antimicrobial resistance (AMR). Therefore, the objective of this work was to evaluate the whole genome of multidrug-resistant <i>E. coli</i> strains isolated from human clinical, animal, and environmental sources. Four <i>E. coli</i> strains previously isolated from human urine (n = 2), retail meat (n = 1), and water from the Rio Grande River (n = 1) collected in northern Tamaulipas, Mexico, were analyzed. <i>E. coli</i> strains were evaluated for antimicrobial susceptibility, followed by whole genome sequencing and bioinformatic analysis. Several ARGs were detected, including <i>bla</i>_CTX-M-15, <i>bla</i>_OXA-1, <i>bla</i>_TEM-1B, <i>bla</i>_CMY-2, <i>qnrB</i>, <i>catB3</i>, <i>sul2</i>, and <i>sul3</i>. Additionally, plasmid replicons (IncFIA, IncFIB, IncFII, IncY, IncR, and Col) and intact prophages were also found. Insertion sequences (ISs) were structurally linked with resistance and virulence genes. Finally, these findings indicate that <i>E. coli</i> strains have a large repertoire of resistance determinants, highlighting a high pathogenic potential and the need to monitor them.