Historically occurring throughout the southeastern United States, the Florida panther is now restricted to less than 5% of its historic range in one breeding population located in southern Florida. Using radio-telemetry data from 87 prime-aged (≥3 years old) adult panthers (35 males and 52 females) during the period 2004 through 2013 (28,720 radio-locations), we analyzed the characteristics of the occupied area and used those attributes in a random forest model to develop a predictive distribution map for resident breeding panthers in southern Florida. Using 10-fold cross validation, the model was 87.5 % accurate in predicting presence or absence of panthers in the 16,678 km2 study area. Analysis of variable importance indicated that the amount of forests and forest edge, hydrology, and human population density were the most important factors determining presence or absence of panthers. Sensitivity analysis showed that the presence of human populations, roads, and agriculture (other than pasture) had strong negative effects on the probability of panther presence. Forest cover and forest edge had strong positive effects. The median model-predicted probability of presence for panther home ranges was 0.81 (0.82 for females and 0.74 for males). The model identified 5579 km2 of suitable breeding habitat remaining in southern Florida; 1399 km2 (25%) of this habitat is in non-protected private ownership. Because there is less panther habitat remaining than previously thought, we recommend that all remaining breeding habitat in south Florida should be maintained, and the current panther range should be expanded into south-central Florida. This model should be useful for evaluating the impacts of future development projects, in prioritizing areas for panther conservation, and in evaluating the potential impacts of sea-level rise and changes in hydrology.