State Route 335 is a north–south state highway in the southern portion of the U.S. state of Ohio. Its southern terminus is at U.S. Route 52 in the village of Sciotoville, part of Portsmouth, and its northern terminus is at State Route 220 in Waverly where it’s concurrent with U.S. Route 23 and State Route 104 for about 1/3 of a mile.

Along the way, it intersects with Ohio 139 in Minford and Ohio 776 near Stockdale. It crosses Ohio 32 near Beaver.

During its concurency with U.S. 23 and State Route 104 in Waverly, its signage is backwards, hence where it joins 23, 335 North becomes 335 South, and 335 South becomes North. For example, The signs during the concurrency should read 23/104 North, 335 South, but instead they all say North.