Several cDNAs encoding the essential human splicing facor (SF) 1 have been cloned. Comparison of the cDNA sequences suggested that the corresponding mRNAs are generated by alternative splicing from a common pre-mRNA. To confirm this assumption and to analyze possible modes used in the generation of these mRNAs, we have determined the structure of the gene encoding SF1. The gene extends over approximately 15kb and contains 14 exons. The exon/intron structure and sequences at the splice sites are highly conserved in the corresponding mouse gene. The human SF1 gene is located on chromosome 11 close to the gene encoding Menin, recently identified as the gene responsible for multiple endocrine neoplasia-type 1 (MEN1). The absence of a TATA box in the 5' flanking region of the SF1 transcription unit suggests that the SF1 gene represents a housekeeping gene. However, genomic sequence analysis revealed putative binding sites for regulatory transcription factors upstream of the 5' end of the cDNA. Analysis of the SF1 genomic and cDNA sequences predicts the use of duplicated 5' and 3' splice sites as well as exon skipping and intron inclusion to generate six SF1 mRNAs by alternative splicing events.