In this study, we identified and characterized a mouse brain-enriched unigene (msvop) after performing digital differential display. Msvop was the mouse ortholog of Xenopus synaptic vesicle-2-associated protein (svop), the molecular characteristics of which were unknown. The 3125-bp full-length cDNA encoded a 548-aa protein of approximately 60 kDa. A strong promoter element was found in the -200 to -100 bp region in both NG108-15 and HEK293 cells. RT-PCR and in situ hybridization analysis confirmed that msvop was strictly expressed in the mouse central nervous system. In adult brains, msvop was highly expressed in the hippocampus and cerebellum. When the gene was transfected into HEK293 cells in a GFP fusion vector, the protein was specifically localized in the cytosol. These results indicate that msvop is a central nervous system-specific gene and could be utilized for elucidating gene regulation in neuronal cells.