Interstate 65 runs from I-10 in Mobile, Alabama, to US 12 and US 20 in Gary, Indiana, just past its interchange with the Indiana Toll Road (I-90). The FHWA route log (routes as designated on December 31, 2025) gives it 884.97 miles: 366.09 in Alabama, 120.13 in Tennessee, 137.32 in Kentucky and 261.43 in Indiana. That total includes 5.73 miles shared with other Interstates: 1.00 with I-40 and 2.60 with I-24 in Nashville, and 2.13 with I-70 in Indianapolis. I-65 does not run with I-90 or I-94. The major cities on the route are Mobile, Montgomery, Birmingham, Nashville, Bowling Green, Louisville, Indianapolis, Lafayette and Gary, and the only tolls are on the Ohio River bridges at Louisville.
At Louisville a truck has tolls and a long work zone to plan for. RiverLink tolls both I-65 bridges over the Ohio: the Abraham Lincoln Bridge carries northbound traffic and the John F. Kennedy Memorial Bridge southbound. From July 1, 2026, rates run from $2.79 for a passenger car to $16.62 for a truck with five or more axles. The rate depends on the vehicle's size and on whether the driver has a prepaid account and a transponder. You do not need an account to cross: RiverLink photographs the plate, and the toll is paid by phone, online or from an invoice mailed to the owner. South of downtown, KYTC is replacing or rehabilitating nine bridges on the I-65 Central Corridor, the five miles between the Watterson Expressway (I-264, exit 131) and Jefferson Street. It closed that stretch on June 1, 2026 and reopened it on July 29 with two lanes each way, a lower speed limit on part of it and some ramps still closed. Work has started on the last six bridges, around the downtown stretch known as Hospital Curve, and KYTC's project site expects them to be done in late 2028. KYTC sends through traffic on a signed detour by I-264 west (Georgia Davis Powers Expressway) and I-64. Kentucky law keeps truck tractors, trailers and semitrailers out of the leftmost lane wherever a road has three or more marked lanes in one direction (KRS 189.340(7)(c)).
Alabama has 366 of the miles. Between Mobile and Baldwin counties, the General W.K. Wilson Jr. Bridge carries four lanes of I-65 for more than six miles across the Mobile-Tensaw Delta. ALDOT inspects it every year, and the 2026 inspection, which began on March 30 and was planned to last eight months, came with warnings of possible lane closures and reduced speeds. Before a hurricane, ALDOT can reverse southbound I-65 so that both sides run north from Mobile to Montgomery. The governor orders it, ALDOT staffs every exit from exit 31 to exit 167, and it has not been needed since Hurricane Dennis in 2005. Hyundai's Montgomery plant is on Hyundai Boulevard, next to exit 164, and I-85 starts at exit 171. In Shelby County, ALDOT is widening I-65 to six lanes between exit 231 near Calera and exit 238 at Alabaster, with completion projected for spring 2028. Around Birmingham, I-459 leaves at milepost 250 in Hoover and I-20/59 crosses at exit 261.
In Nashville, I-65 meets I-440 at exit 80, shares exits 82 to 84 with I-40 and exits 86 to 88 with I-24 (TDOT). Tennessee keeps truck tractors and semitrailers in the right two lanes where signs are posted, except when passing (Tenn. Comp. R. & Regs. 1680-10-02-.03). Winter storms can block the road farther north: in the December 23, 2022 storm, KYTC reported I-65 northbound blocked at mile marker 40 in Warren County, Kentucky, by a jackknifed tractor-trailer. Indiana numbers its exits by INDOT reference post. In Indianapolis, I-65 crosses I-465 at 106 and 123 and runs with I-70 between 110 and 112. INDOT's I-65 Safety and Efficiency project on the southeast side, from north of I-465 to just south of Washington Street, began in spring 2025 and is expected to last about two years. It will leave four lanes each way between I-465 and I-70. At Gary, I-65 crosses I-80/94 (the Borman Expressway) at reference post 259 and meets I-90 at 262, two separate interchanges, before it ends.
- 884.97 miles: 366.09 in Alabama, 120.13 in Tennessee, 137.32 in Kentucky and 261.43 in Indiana, including 5.73 miles shared with other Interstates (FHWA route log, routes as designated on December 31, 2025)
- Southern end at the I-10 interchange in Mobile, AL (milepost 0). Northern end at US 12/US 20 in Gary, IN, just past the Indiana Toll Road (I-90) at reference post 262 (ALDOT; INDOT)
- Shares 1.00 mile with I-40 (exits 82 to 84) and 2.60 miles with I-24 (exits 86 to 88) in Nashville, and 2.13 miles with I-70 (reference posts 110 to 112) in Indianapolis. No overlap with I-90 or I-94 (FHWA route log; TDOT; INDOT)
- The only tolls are on the RiverLink bridges over the Ohio at Louisville: the Abraham Lincoln Bridge northbound and the John F. Kennedy Memorial Bridge southbound. From July 1, 2026, $2.79 for a passenger car up to $16.62 for a truck with five or more axles (RiverLink)
- I-65 Central Corridor, Louisville: two lanes each way between I-264 (exit 131) and Jefferson Street since July 29, 2026. KYTC expects the first three new bridges in March 2027 and the other six, around Hospital Curve, in late 2028 (KYTC)
- Kentucky bars truck tractors, trailers and semitrailers from the leftmost lane where a road has three or more marked lanes in one direction (KRS 189.340(7)(c)). Tennessee keeps them in the right two lanes where signs are posted (Tenn. Comp. R. & Regs. 1680-10-02-.03)
- Hurricane contraflow: on the governor's order, ALDOT can run southbound I-65 northbound from Mobile to Montgomery, with crews at every exit from 31 to 167. It has not been needed since Hurricane Dennis in 2005 (ALDOT)
- Interstate junctions: I-165 (AL milepost 9), I-85 (AL 171), I-459 (AL 250), I-20/59 (AL 261), I-22 (AL, north of downtown Birmingham), I-565 (AL 340), I-840 (TN 59), I-440 (TN 80), I-165 (KY 20), I-265 (KY 125), I-264 (KY 131), I-64 (KY 137), I-265 (IN 6), I-465 (IN 106 and 123), I-865 (IN 129), I-80/94 (IN 259) and I-90 (IN 262) (ALDOT; TDOT; KYTC; INDOT; FHWA)
- Indiana weigh stations at Seymour (reference post 51) and Lowell (241); rest areas at Henryville (22), Taylorsville (71 northbound, 73 southbound), Lebanon (148 northbound, 149 southbound) and Kankakee (231) (INDOT)