In the current study, we isolated and characterized a virulent Siphoviridae phage, designated as pSf-1, which infects S. flexneri. pSf-1 had an efficient bacteriolytic activity against S. flexneri, Shigella boydii, and Shigella sonnei. We also evaluated the biological properties and complete genome sequences of the phage, and the results could play a role in research into the use of phages against shigellosis.