The state of Kentucky is the setting of Kentucky Route Zero, where it takes the form of the overworld map. Most of the game's locations are located off of the various highways that crisscross the map, and as such the protagonists are often found traveling its roads, notably Interstate 65. Notable locations include Equus Oils, Marquez Farmhouse, Elkhorn Mine, and The Lower Depths. While traversed in-game, it is represented by a black and white highway map.

The in-game map of Kentucky

The map is fairly accurate to real life; it represents a large stretch of Interstate 65 between Bowling Green and Elizabethtown. This places the Equus Oils station somewhere in the vicinity of Smiths Grove, and the Marquez Farmhouse just west of Cave City. The empty area in the center of the map is occupied by Mammoth Cave National Park. Areas beyond the roads, like Lake Cumberland, are also correctly represented.