Immediately after this concurrency terminus, I-565 is featured in another junction as downtown Huntsville rises into view to the east.

It passes by the hub of the Huntsville Hospital system, which runs throughout North Alabama, and begins its journey across Monte Sano mountain.

After leaving the mountain range, it has no more major junctions past this point as it loses the Governor's Drive name.

After about 20 miles (32 km) of hilly terrain, the route crosses the Tennessee River and enters Guntersville, junctioning with SR 79 north.

The route then climbs and descends multiple hills as it enters into Alexandria, where it junctions with SR 144.

It eventually climbs up a steep hill and gives drivers a great view of Anniston, Cheaha State Park, and some of the highest mountains in Alabama as it descends a steep grade into Anniston.

The route merges with I-20 and about two miles (3.2 km), it leaves it, turning south, and immediately enters Cleburne County.

The route junctions again with SR 165 and crosses a fork of the Chattahoochee River.

Just before the route can cross the river into Georgia, it loses US 82 and moves away from the state line.

It loses US 84 and continues southwest independently with SR 210, the Ross Clark Circle.

ALDOT has prioritized four-laning the route in Alabama as a viable north–south road corridor in the eastern part of the state.

Of particular concern was the segment from Seale south to the Barbour County line, where rolling hills along the original alignment limit the visibility of oncoming traffic, contributing to poor decisions by motorists to pass, resulting in numerous head-on collisions.

In this segment alone, 31 people were killed in crashes between 1992 and 2006, leading Reader's Digest to proclaim it one of "America's Deadliest Highways" in 2000.

[3] After being contacted by and subsequently meeting the family of a fatal crash victim in 2003, ALDOT Director Joe McInnes decided to expedite the construction on the last remaining stretch of two-lane road between Seale and the Barbour County line.

Then, after diverting from I-20 east of Oxford at exit 191, it reverts to a two-lane highway southward to Opelika, with a brief four-lane stretch just south of Wedowee.

From Opelika southward to its southern terminus in Dothan, US 431 is now open as a four-lane highway.

Despite being planned for years, construction did not begin until after receiving funding in a 2009 economic stimulus bill.

[6] Prior to the start of construction, archeological work on part of the route that passed through Fort McClellan located Native American spearpoints and an American Civil War homestead.

The northern portion, which began construction in 2010,[8] is built with two at-grade intersections (McClellan Bypass and Summerall Road) and one trumpet interchange (SR 21/McClellan Boulevard).

Much development has been the result of this highway in the past decade along I-20 in Oxford and it is regarded as a way to redevelop McClellan as well as north Anniston.