Escherichia coli species exhibits a high genomic diversification from evolution, mobile genetic elements and recombination. An environmental E. coli isolate, 'JHI_5025' from a crop trial appeared to be clonally related to the historical reference isolate E. coli K-12 strain 'MG1655', warranting further genomic analysis. Their genomes share an average nucleotide identity of 99.74% and whole genome alignment showed little rearrangement of the JHI_5025 sequence compared to the reference. Five genomic islands not in the reference aligned to other sequences in the Enterobacteriaceae. Isolate JHI_5025 contained E. coli K-12 F plasmid sequence and at least one complete prophage sequence. The genome and comparison dataset provides utility of E. coli JHI_5025 as a representative contemporary genetic mimic of a well-known and much used workhorse strain.