Shiga toxin-producing Escherichia coli (STEC) is an important pathogen that can cause asymptomatic infections, diarrhea, hemorrhagic colitis (HC), and life-threatening hemolytic uremic syndrome (HUS) in humans. Shiga toxins (Stxs) are the major virulence factors encoded by prophages, which play a crucial role in STEC pathogenesis and evolution. In this study, seven Stx phages were obtained from STEC isolates derived from four asymptomatic food handlers, two diarrheal patients, and one outbreak-related HUS case in China. These phages exhibited three morphologies: an icosahedral head with either a short or a long tail, and an elongated head with a long tail. Of these seven phages, three were sequenced; two showed a complete identity with their respective prophage sequences, while phage phiXuzhou21-Stx2a lacked a 6011 bp region-encoding integrase, excisionase, and hypothetical proteins. Comparative genome analysis revealed that the induced seven phages primarily varied in their regulatory regions, whereas the short-tailed phages showed high similarity in their morphogenesis-related regions. In addition, five of the seven phages demonstrated the ability to convert non-pathogenic E. coli strains into Stx-producing transduced strains. Under inducing conditions, Stx expression levels were significantly increased in these transduced strains. These findings underscore the diversity and adaptability of Stx phages and emphasize the importance of understanding their genetic and molecular interactions with host bacteria.