Approximately 30 kb of the mouse genome, containing the gene for a major seminal vesicle transcript, has been cloned. The gene was identified by the similarity to members of a family with rapidly evolving genes that includes the gene encoding the major clot protein in rat semen, SVS II, and the human semenogelin genes. The nucleotide sequence of 16.9 kb was determined; this sequence encompasses the gene of 2215 bp plus 9-kb and 5.6-kb regions flanking the 5' and 3' ends of the gene. The transcription unit is divided into three exons, of which the first encodes the signal peptide, the second the secreted protein, while the third exon contains 3'-nontranslated nucleotides only. The transcript encodes a protein of 375 amino acid residues, including a signal peptide of 22 residues. The secreted polypeptide is a protein of Mr 38442 and is similar in sequence but smaller than the major clot-forming protein of rat semen, SVS II. It is highly charged at pH 7 and it has an isoelectric point of 10.68. The central part of the protein consists of tandem repeats that might serve as a substrate for transglutaminase.