The highway's southern terminus is in the Brickell neighborhood of Downtown Miami at an intersection with Brickell Avenue (US 1), and its northern terminus is east of Copper Harbor, Michigan, at a modest cul-de-sac near Fort Wilkins Historic State Park at the tip of the Keweenaw Peninsula.

In Florida, US 41 is paralleled by Interstate 75 all the way from Miami to Georgia (on the northern border), and I-75 has largely supplanted US 41 as a major highway.

Between Miami and Naples, US 41 is signed east–west and cuts across the Florida peninsula, running through the vast Everglades wilderness.

The highway proceeds through South and Central Georgia cities and towns including Adel, Tifton, Cordele, Unadilla, Perry, Warner Robins, Macon, Barnesville, and Griffin as it approaches the Atlanta metropolitan area.

In Atlanta, Highway 41 was formerly carried on Spring Street near Five Points, but it has long been re-routed via Northside Drive around the downtown area.

The Mercedes-Benz Stadium, Georgia World Congress Center, and State Farm Arena are located off Northside Drive.

US 41 also passes through Marietta, Kennesaw, Acworth, Cartersville, Adairsville, Calhoun, and Dalton en route to Tennessee.

It is called Ringgold Road through East Ridge up to the Bachman Tunnel, where it enters Chattanooga and then around the base of Lookout Mountain.

It then heads through the towns of Lookout Valley, Jasper, and other communities before ascending the Cumberland Plateau, running through Tracy City and Monteagle,[4] where it descends toward Manchester.

At US 60, US 41 becomes a four-lane surface road with stoplights as it crosses through the northern parts of Henderson and John James Audubon State Park.

Due to natural fluctuations of the Ohio River's path, US 41 remains in Kentucky for about one mile north of the Twin Bridges.

However north of Terre Haute, US 41 becomes a secondary road, passing through smaller towns such as Rockville and Attica on the east side of the Wabash River.

State Road 63 is the main route north of Terre Haute in this area since it is a four-lane highway on the west side of the Wabash River.

US 41 returns to a four-lane divided highway just south of Boswell where SR 63 ends, staying as such until reaching Cedar Lake.

From Cedar Lake north to the Illinois state line, US 41 is a well-travelled road with numerous stoplights, with the exception of the mile-stretch where it overlaps Interstate 80/94.

The three US routes run together northwest along Indianapolis Boulevard then cut north on Ewing Avenue on the south side.

US 41, as Lake Shore Drive, passes by the Museum of Science and Industry (at 57th Street) in the Hyde Park area.

The roadway then has a sharp S-curve called the Oak Street Curve where the suggested speed limit is 25 mph (40 km/h).

In the northern suburbs of Chicago, US 41 joins I-94 (Edens Expressway) just north of Lake Avenue for a short distance before splitting from the freeway just south of Lake–Cook Road and continuing north as the Skokie Highway for roughly 25 miles (40 km) to a point near the Wisconsin border.

US 41 then heads north-northwest on a freeway to Fond du Lac and Oshkosh, then northeast to Appleton and Green Bay.

US 41 is a freeway for nearly 70% of its length through Wisconsin, with the exceptions being an expressway section north of Green Bay and surface streets in Marinette.

I-41 has been cosigned with US 41 along the freeway sections between a point just south of the Illinois state line where US 41 and I-94 split, and the I-43 interchange northwest of Green Bay.

The trunkline ends at a cul-de-sac east of Fort Wilkins Historic State Park in Copper Harbor.

[12] Historical landmarks along the trunkline include the Marquette Branch Prison, Peshekee River Bridge, and the Quincy Mine.

In 1953, US 41 was extended along US 1 and State Road A1A (SR A1A) to terminate in Miami Beach, Florida, but it was truncated back to the earlier terminus in 2000.

[19] Those efforts came to fruition in April 2015, almost ten years later, with US 41 signage mostly replaced by I-41 starting at the Illinois state line, following I-894 around the west end of Milwaukee, and on to Green Bay.