In urbanized landscapes, golf course ponds may provide the only remaining habitat for semi-aquatic animals. Eastern Mud Turtles (Kinosternon subrubrum), which rely heavily on both aquatic and terrestrial habitats, may face challenges on golf courses, which typically have significantly modified and fragmented landscapes. We conducted a radio-telemetric study of 11 mud turtles inhabiting a golf course pond in the western Piedmont of North Carolina to investigate their terrestrial activity and habitat selection in a fragmented landscape. Most turtles moved to terrestrial habitats in late summer and emigrated a mean distance (± SE) of 187.2 ± 67.4 m and moved a mean straight line distance (± SE) of 119.3 ± 47.4 m from the pond. We determined habitat selection using logistic regression to compare turtle locations with random locations and found that mud turtles selected forested habitats with moderate canopy cover and no grass. Mud turtles also selected habitat containing herbaceous vegetation and woody debris as overwintering locations. Mud turtles did not select heavily disturbed habitats with limited canopy cover and pavement or cut grass associated with fairways, roughs, and residential lawns. Overall, our study suggests that maintaining relatively undisturbed forested habitat within fragmented urban landscapes, such as those found on golf courses, may allow for the persistence of these semi-aquatic turtles. Information from this study can be used to better understand critical upland habitat requirements of other semi-aquatic species inhabiting fragmented landscapes and aid in the implementation of habitat management plans.