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E. coli sequence type 131 (ST131) is one of pathogens that causes resistant infections. Comparative genome analyses allow interpretations of the virulence factors of pathogens. Thus, in this study, we analysis the genomic differences between the pathogenic E. coli ST131 and the non-pathogenic E. coli K-12. In this study, we identify the genomic differences between 96 E. coli ST131 strains and the E. coli K-12 in gene elements and their non-coding regulation elements. Using next-generation whole-genome sequencing data, we investigated genetic variations of protein-coding regions and their regulation regions. After the alignment of the sequence reads, large numbers of single nucleotide variants (SNVs) were observed in the regulation and protein-coding sequences. In the regulation regions, we found strong conserved regions, in this case, ribosome binding sites. In the gene regions, we found conserved start and stop codons with the specific position varying commonly in each codon. Except for these well-conserved regions, other variations were randomly distributed in regulation regions. Even a region having well-known conserved sequences such as -10 and -35 in the promoter had a similar level of variation. In this study, we found genomic variations between the pathogenic E. coli ST 131 strain and the non-pathogenic E. coli K-12. In addition, the numbers of sequence variations were determined in both the protein-coding regions and the regulation regions. However, we found that the effects of variations on the protein-coding regions are less significant than those on the regulation regions.