The murine urokinase-type plasminogen activator receptor (uPAR) gene has been isolated and its complete nucleotide sequence established. The gene is organized into seven exons comprising 9.5% of the 13,207-base pair region that spans the interval between the transcription initiation and polyadenylation sites. The region upstream of the transcription initiation site lacks TATA- or CCAAT-like elements but is flanked by a G+C-rich region, which contains a number of potential regulatory elements including Sp1 and AP1 binding motifs. The close association of both Sp1 and AP1 sites within the proximal promoter region is consistent with the observation that the murine uPAR gene is inducible by phorbol esters. The major functional domains of the encoded protein, including the signal peptide, three cysteine-rich internal repeats, and the glycolipid anchor attachment motif, are encoded by separate exons. Based on the organization of the murine uPAR gene and the distribution of protein domains within the exons in the Ly-6 family of genes, it appears that the uPAR gene evolved secondarily to two internal duplication events within a Ly-6-like ancestral gene. The cloned and sequenced murine uPAR gene will be a valuable tool in understanding the regulation and biological roles of uPAR in that it will permit detailed studies of gene expression and uPAR-dependent processes in vitro, as well as the generation of both gain-of-function and loss-of-function mutants in transgenic mice.