Objective:A novel multidrug resistant(MDR)Escherichia coli(E.coli)strain J2_5 was isolated from a patient’s mid ⁃ stream urine sample. According to the genome sequence of J2_5,we analyzed its evolutionary classification,drug resistance genes and virulence genes,providing theoretical basis for clinical diagnosis and drug selection for drug resistant bacterial infection. Methods: Genome DNA of J2_5 was isolated and sequenced by high ⁃ throughput sequencing technology,after analyzed the high ⁃ throughput sequencing data and compared it with known databases by a variety of bioinformatics tools,a high integrity genome of J2_5 was obtained. Results:The total length of the whole genome of J2_5 was 4.7Mb and GC content was 50.78%. Phylogenetic tree analysis showed that J2_ 5’genome was closely related to that of wild type E. coli strain MG1655. But Multilocus sequence typing(MLST)analysis revealed ST type of J2_5 was ST2491,which was different from that of MG1655(ST10). Furthermore,CARD database analysis showed that the genome of J2_5 encodes 65 drug resistance genes,which make the strain obtain drug⁃resistant ability through four different mechanisms (antibiotic efflux pump,antibiotic inactivation,antibiotic target change,reduced permeability to antibiotics). VFDB database analysis showed that there were 40 virulence genes in 7 categories,encoding adhesive proteins,invasion proteins,autonomic transporters and other toxin proteins. After analyzing the whole genome of J2_5 through the Prophage Hunter online tool,we found that there were three prophages in the genome of J2_5,and a virulence gene was carried by one of the prophages. Conclusion:The whole genome sequencing results showed that J2_5 carries some virulence genes and drug resistance genes,which makes the strain obtain strong pathogenicity and resistance ability to a variety of antibiotics.