We deleted the essential gene 24 from the genome of bacteriophage T4. The delta 24 phage is a conditional lethal mutant that can grow only when the host strain supplies the product of gene 24 in trans, or when the phage acquires a functional gene 24 by some type of recombination event. Thus, gene 24 can be used as a selectable marker, for example permitting transposition into the T4 genome and analyses of plasmid-phage recombination [Woodworth and Kreuzer, Mol. Microbiol. 6 (1992) 1289-1296; H.W.E. and K.N.K., manuscript submitted]. We also found that the promoter region of gene 24 allows a low level of autonomous plasmid replication in T4-infected cells, raising the possibility of a previously unrecognized mode of T4 replication initiation.