U.S. Route 301 runs north–south from the Savannah River southwest of Allendale to Hamer approaching the South of the Border roadside attraction complex.

US 301 enters South Carolina over the Savannah River southwest of Allendale. Concurrencies include U.S. 321 in and around Ulmer, U.S. 601 between Bamberg and Orangeburg, then I-95/U.S. 15 Santee, at Exit 97 before all three move across Lake Marion. The U.S. 15-301 multiplex leaves I-95 at Exit 102, running in close proximity to I-95, but U.S. 15 breaks away at Summerton. Other concurrencies include U.S. 521 in Manning, U.S. 52 between Effingham and just south of Florence, U.S. 76 eastbound from east of Florence until Pee Dee, and finally joins U.S. 501 in Latta, where they both approach the South of the Border roadside attraction complex at the interchange with I-95 on the north–South Carolina border.

US 301 was established in 1932 as a replacement of the piece of US 17-1 north of Wilson and the whole of US 217. Thus US 301 initially ran from US 17 (now US 76) at Pee Dee northeast through Dillon, South Carolina, into North Carolina, and Virginia, ending at U.S. Route 1 in Petersburg, Virginia. This entire route is now paralleled by Interstate 95.

In 1935, US 301 was extended southwest to U.S. Route 15 at Summerton, South Carolina. This extension took it west on U.S. Route 76 (formerly U.S. Route 17) to Florence, South Carolina and south on U.S. Route 52 (also formerly US 17) to Effingham, South Carolina. There it split to the southwest, running along what had been S.C. Highway 4 via Manning, South Carolina to Summerton. This again runs parallel to Interstate 95.

In the late 1940s, US 301 was extended again, south all the way towards Tampa, Florida. On the way there from South Carolina it ran along U.S. Route 15 southwest to Santee, South Carolina and replaced S.C. Highway 4 west to Orangeburg, South Carolina. From Orangeburg, US 301 ran southwest with U.S. Route 601 to Bamberg, South Carolina, replaced S.C. Highway 33 to Ulmer, South Carolina, replaced S.C. Highway 508 to Allendale, South Carolina, and replaced S.C. Highway 73 to the Georgia state line. When Interstate 95 was built across Lake Marion in 1968, US 301 and 15 ran parallel to it until they were relocated in a triple concurrency in 1987.

Bannered routes of U.S. Route 301