Understanding the spatial ecology of an invasive species is critical for designing effective control programs. Determining and quantifying home range estimates and habitat associations can streamline targeted removal efforts for wide-ranging, cryptic animals. The Burmese python (Python bivittatus) is a large-bodied constrictor snake with an established and expanding invasive population in southern Florida. This apex predator has severely impacted native wildlife across the Greater Everglades ecosystem. However, limited ecological information exists on this invasive species at the landscape level. Here, we present results from a study using radiotelemetry to quantify movements and habitat use patterns of 25 adult Burmese pythons in southwestern Florida, USA, for average periods of 814 d (range: 288–1809). Our objective was to quantify home range size, movement rates, and second- and third-order habitat selection. Mean annual home range size was 7.5 km² ± 2.9 km² (95% kernel density estimate), and pythons moved at a maximum mean daily rate of 0.52 km/d. Burmese pythons selected agriculture, freshwater wetland, saline wetland, and upland land cover classes but avoided open water and urban land cover classes. Nest site selection was highest for pythons at an elevation of 1.7 m with nesting hotspots concentrated on the borders of urban and agricultural areas or in sandy forested upland habitats. A broader understanding of the spatial utilization of Burmese pythons will enhance the utility of emerging control strategies across their invaded range.