The portion of the former William H. Natcher Parkway between US 231 and I-65 is not a part of the Interstate Highway System as per federal regulations and is designated as Kentucky Route 9007 (KY 9007).

I-165 travels along the west side of the city, and continues in a northwesterly direction through rolling farmlands and near coal mines for 70.2 miles (113.0 km) before meeting its northern terminus at an interchange with US 60 in Owensboro.

I-165 traverses Warren, Butler Ohio, and Daviess Counties, bypassing the cities of Morgantown, Beaver Dam, and Hartford.

[5] From July to August 2017, construction consisting of shoulder widening, drainage upgrades, and repaving was completed.

Traffic flow was restricted to one lane, wide loads were prohibited, and the speed limit set to 55 mph (89 km/h).