It runs through Holy Trinity, Alabama in the eastern part of the state.

Its northern terminus is at U.S. Route 431 (US 431) south of Phenix City, Alabama, and its southern terminus is at US 431 5 miles (8.0 km) north of Eufaula, Alabama.

It travels near Eufaula National Wildlife Refuge before it enters Russell County and intersects SR 208 near Cottonton.

SR 165 then goes through Holy Trinity and Fort Mitchell as it goes near the Fort Mitchell National Cemetery and terminates at US 431 near the Columbus, Georgia metropolitan area.

[1][2] While a rural highway, traffic counts are quite high due to MeadWestvaco operations at Cottonton, and because it is a frequent route for tourists headed to the Florida Gulf Coast.