It begins at an interchange with U.S. Route 151 (US 151) in Anamosa and ends at the Dale Gardner Veterans Memorial Bridge over the Mississippi River near Sabula.

Originally, Iowa 64 spanned the length of the state and began at the Missouri River in Council Bluffs where it connected to N-64 in Omaha.

After an S curve that takes the road to the south and then back east, the highway passes Antioch School, which Iowa painter Grant Wood attended for four years.

Iowa 64 passes through the small towns of Monmouth and Baldwin and bypasses the village of Nashville before arriving in Maquoketa, the seat of Jackson County.

North of Sabula, the US 52 / Iowa 64 causeway divides the Mississippi River from Sheepshead Bay, another backwater area.

US 52 / Iowa 64 turn east and cross the main channel of the Mississippi River on the Dale Gardner Veterans Memorial Bridge, becoming US 52 / Illinois Route 64 (IL 64).