The highway is largely parallel to I-587 between Wilson and Greenville and primarily serves smaller communities such as Saratoga and Farmville.

East of Greenville, US 264 is an important highway connecting communities such as Washington, Belhaven, Swan Quarter, and Engelhard.

Prior to the establishment of US 264, North Carolina Highway 91 (NC 91) running along the general corridor between Zebulon and Engelhard.

In 1932, the American Association of State Highway Officials (AASHO) approved the establishment of US 264 between Zebulon and Engelhard, running concurrently with NC 91.

US 264 was subsequently removed from the freeway between Wilson and Greenville and was placed along portions of its old routing and a concurrency with NC 11 Bypass.

Interchanges with Smithfield Road, Wendell Falls Parkway, and US 64 Business provide access to the town.

The highway enters northern Zebulon from the west, and meets NC 96 at an interchange, before diverging from US 64 and travelling to the southeast.

East of the interchange, the highway leaves the town limits of Zebulon and travels southeast through rural Wake County.

US 264 then crosses Turkey Creek and Big Branch before running north of Bailey where it meets NC 581.

The highway meets Green Pond Road one-half mile (zero point eight zero kilometres) east of the Nash County–Wilson County line.

The highway then crosses over a railroad owned and operated by the Carolina Coastal Railway before meeting I-95, I-587, and I-795 at a cloverleaf interchange.

[1][3] From I-587, US 264 travels through rural Wilson County for 1.7 miles (2.7 km) until reaching the town limits of Saratoga and picking up the name Main Street.

At the intersection, US 264 turns southeast to run concurrently along NC 121 for approximately one-half mile (zero point eight zero kilometres).

NC 121 then turns to follow Wilson Street towards Farmville while US 264 continues southeast and enters into Pitt County.

US 258 follows concurrently with US 264 for approximately one-half mile (zero point eight zero kilometres) until turning to the northeast along Wesley Church Road.

US 264 turns to the east at the intersection travelling along a non-controlled access divided highway which somewhat parallels the Tar River.

[1][3] As US 264 continues east on Pactolus Highway, it leaves the city of Greenville and reenters rural Pitt County.

US 264 then turns to the south and travels approximately 4 miles (6.4 km) east of NC 30 until crossing Tranters Creek, a tributary of the Tar River, and entering Beaufort County.

US 264 continues east for 5.5 miles (8.9 km) until crossing another tributary of the Pungo River and entering into Hyde County.

US 264 and NC 45 continue southeast for 2.4 miles (3.9 km) until turning to the south and crossing the Intracoastal Waterway along the Walter B. Jones Bridge.

[5] The highways continue south for approximately seven miles (eleven kilometres) through rural Hyde County passing the community of Scranton to the east.

On the north side of the bay, it intersects Bayview Drive which provides access to the community of Stumpy Point.

US 264 continues north through the Alligator River National Wildlife Refuge for 5.6 miles (9.0 km) before turning to the northwest.

US 264 travels for an additional 2.5 miles (4.0 km) before intersecting its eastern terminus at US 64 west of Manns Harbor in Dare County.

[7] In 2016, construction began on a bypass between US 264 in Greenville, where it connects with the city's Stantonsburg Road, to NC 11 as it passes through Ayden.

[13] In June 2022, after approval from AASHTO in 2021 Annual Meeting, mainline US 264 was rerouted back along its former routing between north of Saratoga to Greenville with US 264 Alternate decommissioned along that section.

[18][7][13] Established around 1954 as a renumbering of mainline US 264, it traversed through downtown Farmville, via Wilson Street and Moye-Turnage Road.

[21][22][23][24] Established by 1954 as a renumbering of mainline US 264, it traversed through downtown Greenville via Dickinson Avenue and East 10th Street.

Although no longer maintained after the company went bankrupt in 1861, the route was still used as an unpaved track and Right of way by horse and motor traffic.

State senator James Leonidas Fleming, who established East Carolina University in Greenville, was killed in an early automobile accident along the plank road in 1909.