Interstate 65 (abbreviated I-65) is a controlled-access, interstate highway traversing the eastern United States serving as a connector between Mobile, Alabama and Gary, Indiana, paralleling the older U.S. Highway 31 from Mobile to Indianapolis. Its 887.3 mile (1,401 km) route also takes this major thoroughfare through Birmingham, Alabama; Nashville, Tennesee; Louisville, Kentucky; and Indianapolis, Indiana.
The interstate is the primary north-south route through the state of Alabama, linking the state's four largest cities: Mobile, Montgomery, Birmingham, and Huntsville (via the I-565 spur). In the Birmingham metro area, I-65 serves as the main thoroughfare for traffic traveling north and south. It passes through the communities of Calera, Alabaster, Pelham, Hoover, Homewood, Birmingham, Fultondale, and Gardendale in both Shelby and Jefferson Counties.
 Control cities
Through Birmingham, Huntsville, located 98 miles to the north, is used as the control city for northbound lanes. However, Nashville, Tennessee, was used as the northbound control city through the early 1990s when the switch to Huntsville was made. The switch came after Huntsville was connected to the interstate system by I-565. Montgomery, located 92 miles to the south, is used for southbound lanes.
Birmingham is first used as a control city at Exit 340 in Decatur for southbound travelers. However, through the early 1990s, it was first utilized as a control city in Nashville. For northbound travelers, Birmingham first appears at Exit 171 in Montgomery.
 Major intersections
In Hoover, I-65 intersects I-459 at exit 250 in a large stack interchange. This stack interchange holds the record for the costliest interchange ever constructed within the state of Alabama. It will retain this title until the I-65/I-22 interchange is completed in Birmingham during the early 2010s.
In downtown Birmingham, I-65 intersects I-20/I-59 at exit 261 in a large intersection known locally as Malfunction Junction. The aforementioned I-22 interchange is planned to be constructed between Malfunction Junction and an existing, unique, scrape intersection with U.S. Highway 31 to the north.
The first section of what became I-65 was a 25-mile stretch between a point four miles south of Clanton and a point about two miles north of Calera which was opened on November 26, 1960 just in time for traffic coming to Birmingham for the 1960 Iron Bowl.
The final section of I-65, between exits 266 (Fultondale) and 280 (Kimberly/Warrior), was opened in 1985. The entire section has a minimum of three lanes in each direction. After this section opened, additional lanes were constructed for the existing section south of exit 266. Upon completion of that project, in the late 1990s or early 2000s, several miles of I-65 north of exit 280 were expanded from two lanes in each direction to three.
On July 2, 2002 all of I-65 in Alabama was dedicated as the nation's first "Heroes Highway" in honor of Mike Spann, the first American killed in combat during the US invasion of Afghanistan on November 25, 2001.
In October 2008, Interstate 65 became the nation's first biofuels corridor with E85 ethanol and B20 biodiesel fuels available along its full length. The project was funded with U.S. Department of Energy grant of $1.3 million. Stations in the Birmingham area include Dogwood Shell at 1488 Montgomery Highway in Vestavia Hills and County Line Shell at 313 Cane Creek Road in Warrior.
Interstate 22 from Memphis, Tennessee will connect with I-65 at the new exit 265. Construction on this interchange began on August 1, 2010 and is scheduled for completion in October 2014. Preliminary roadwork to widen I-65 between 16th Street North and 41st Avenue North has been ongoing since 2007. Just a few miles north of I-22 will be a new interchange (exit 274) connecting to the Northern Beltline (future Interstate 422), providing a bypass to I-59 northeast of Birmingham and I-20/59 to the southwest of the city and completing the loop started with I-459 around the south and east.
In Shelby County, work is underway to expand I-65 from four to eight lanes between Alabama Highway 119 (exit 246) and Shelby County 52 (exit 242). When that project is completed, the next widening project will work from 52 to U. S. Highway 31 (1st Street Alabaster, exit 238). A third project will then widen the interstate from there to the Shelby County Airport (exit 234). No dates have been given for the future projects.
 Major accidents
In being a major thoroughfare for both local and interstate travel, accidents are nearly a daily occurrence. Although there are too many to list, there have been several notable accidents in recent years. In both, 2002 and 2004 incidents at Malfunction Junction resulted in partial closure of some travel lanes while bridges were reconstructed.
- "New highway between here, capital to open" (November 24, 1960) Birmingham News - via Birmingham Rewound
- MacDonald, Ginny (November 26, 2006). "Roads chief wants I-22 done by 2010." The Birmingham News
- "Interstate 65" (November 28, 2006). Wikipedia, The Free Encyclopedia - accessed November 29, 2006
- Averette, Justin (August 19, 2008). "State opens bids on I-65 project." Shelby County Reporter
- Cooper, Lauren B. (October 6, 2008). "Consortium opens 13 biofuel stations in Alabama". Birmingham Business Journal
- "Interstate 65 in Alabama" (August 15, 2010) Wikipedia, The Free Encyclopedia - accessed November 1, 2010
- Wagner, Neal. (August 26, 2010). "ALDOT soon to widen I-65 to Promenade exit." Shelby County Reporter - accessed November 2, 1010.