It runs 433 miles (697 km) from Miami in South Florida northwest to the Georgia state line, with the overall route continuing to Tennessee in the Rocky Top area.

Like all AASHTO designated highways in Florida, US 441 always carries a FDOT designated hidden state road number: Concurrences with U.S. routes include US 98 from Royal Palm Beach to Okeechobee, within the vicinity of Lake Okeechobee, US 192 between Holopaw and Kissimmee, US 17-92 from Kissimmee to Orlando, SR 44 from Mount Dora to Leesburg, US 27 from Leesburg to Ocala, US 301 from Belleview to near Sparr, and US 41 from High Springs to Lake City.

The route is concurrent with SR 7 from its southern terminus and parallels Interstate 95 (I-95) north out of Miami to western Aventura, where I-95 heads northeast to access Fort Lauderdale and the remainder of the eastern Florida coast.

Additionally, US 441 meets Interstate 595 (Exits 8 and 8B) west of Fort Lauderdale and the tolled State Road 869 in Coconut Creek.

Just north of Pahokee in Canal Point, SR 700 rejoins US 98-441 and remains concurrent with US 441 along the eastern edge of the lake.

US 441 continues north, meeting the Turnpike at Yeehaw Junction in Osceola County (via State Road 60).

Immediately after entering Orange County, it intersects SR 417 (Central Florida GreeneWay) at exit 11.

Further northwest it runs beneath an interchange with State Road 414, then cuts through a corner of Seminole County and re-enters Orange County in Apopka where it meets the northern terminus of West Orange Trail and the tolled State Road 429 before leaving the Orlando area.

This road was intended to be part of the Central Florida GreeneWay loop around the Orlando Metropolitan Area.

CR Old 441 remains a local downtown street in Mount Dora, but US 441 continues as a rural at-grade four-lane divided highway.

Continuing with the rural surroundings, US 441 maintains its straight north and south position until it curves to the northwest between Lincoln and East Pine Avenues.

Because of the frequent use of street-legal golf cars in the community, bridges and tunnels for these carts can be found throughout the road.

US 27-441 leaves The Villages as it approaches County Road 42, a bi-county east-west scenic route through Central Florida.

The highway reaches the heart of Ocala at the intersection with SR 40 (West Silver Springs Boulevard).

From there it intersects CR 318, which provides access to I-75 in Irvine to the west and Salt Springs in Ocala National Forest to the east.

It passes by a turnoff to Cross Creek before entering Micanopy, then runs across Paynes Prairie Preserve State Park and goes through Gainesville as 13th Street, serving as the eastern edge of the University of Florida campus.

Just as it did in Leesburg, US 441 in High Springs curves to the right while a state road becomes independent, specifically SR 20, which runs west only to join US 27 as they head towards Perry, Tallahassee, and Niceville.

US 441 has an interchange with Interstate 10 (Exit 303), then skirts the western edge of the Okefenokee Swamp and east bank of the Suwannee River as it enters Georgia.

The "shield" for US 441 was brown, until the state was forced by the federal government to conform to standards that required consistent black-and-white signs in 1993.