Bovine adenovirus type 3 (BAV3) is a DNA virus that causes respiratory and gastrointestinal disorders in cattle. The viral genome consists of a linear double-stranded DNA molecule (35,000 base pairs) with inverted terminal repeats at each of its 5’ molecular ends. We have subcloned 10 Hindlll fragments spanning 4.9–96.0%, 5 EcoRl fragments spanning 3.4–89.5% and 2 Xbal fragments spanning 35.7–82.9% of the BAV3 (strain WBR-1) genome into the bacterial cloning vector pUC19. The subcloning of the viral genome facilitated the construction of linear restriction enzyme maps for BamHI, Clal, EcoRl, Hindlll, Kpnl, Notl, NspV, Pstl, Pvul, Sall, Xbal and Xhol. In this study we report on the molecular cloning and restriction endonuclease mapping of the BAV3