State Road 24 runs along a former branch of the Seaboard Air Line Railroad.

The northeast corner of SR 24 and SR 222 (North 39th Avenue) is the site of the Gainesville Regional Airport, while the rest of the road runs through the University of Florida Austin Carey Forest before finally terminating at an interchange with US 301 in Waldo.

Motorists who want to travel in the opposite direction must use nearby Shealy Drive.

It travels 0.76 miles (1.22 km) west of SR 24 to U.S. Highway 27 Alternate and is known locally as Picnic Street.

Both routes follow the interstate until leaving at Exit 382 to make an easterly turn onto a new overlap with State Road 121.

The four truck routes run northeast along SR 331 past the eastern terminus of Florida State Road 226, and are also joined by another overlap, this time with Florida State Road 24A.

This makes a total of five overlapping routes with SR 331, which curves from northeast to the north.