Recent reports of declines in terrestrial snake populations in the southeastern U.S. have highlighted the need for life history information for these often cryptic animals. In this study, we used radio-telemetry to describe home range size and habitat use of the Florida Pine Snake (Pituophis melanoleucus mugitus), a large fossorial species associated with the endangered Longleaf Pine (Pinus palustris) ecosystem. Florida Pine Snakes had large home ranges that varied in size among individuals (mean  =  59.2 ± 50.8 ha, minimum convex polygon). There was no significant difference between mean annual home range size of males and females, but home range size differed significantly among seasons and between males and females by season. At the home range scale, Florida Pine Snakes selected natural pine forests and mature slash pine plantations, and they tended to use sites with shrubs and vines and very little bare ground at a local scale. As has been observed in other studies, Florida Pine Snakes were highly fossorial and they frequently sought shelter in Southeastern Pocket Gopher (Geomys pinetus) burrows. Snakes in our study rarely crossed paved or graded dirt roads; however, they frequently crossed low-use harrowed dirt roads. Collectively, our findings indicate that Florida Pine Snakes require large contiguous pine forests and that paved and graded roads may be significant barriers to their movement.