srfA is an operon required for the production of the lipopeptide antibiotic surfactin, competence development, and efficient sporulation in Bacillus subtilis. The expression of srfA is induced after the end of exponential growth and is dependent on the products of late-growth regulatory genes comP, comA, and spo0K. To begin to understand the mechanism of srfA regulation, the srfA promoter region was identified and characterized. To examine srfA promoter activity, the srfA promoter was fused to lacZ and inserted into the B. subtilis chromosome as a single copy at the SP beta prophage. The location of the transcription start site of srfA was determined by primer extension analysis and shown to be preceded by a sequence that resembles the consensus promoter recognized by the sigma A form of RNA polymerase. The srfA operon was found to have a sequence corresponding to a long, untranslated leader region of the srfA mRNA (300 bp). A nucleotide sequence and mutational analysis of the promoter identified a region of dyad symmetry required for srfA-lacZ expression. A similar sequence is found in the region upstream of the degQ promoter, transcription from which is also regulated by ComA. This region of dyad symmetry found upstream of these promoters may be the target for ComA-dependent transcriptional activation.