KY 101 extends through three geophysical subregions of the Interior Low Plateaus, which stretch from Tennessee to north of the Ohio River.

In Warren County, the highway crosses the Western Pennyroyal Karst Plain, a valley that features innumerable ponds and sinkholes.

KY 101's Edmonson County stretch is through the Mammoth Cave Uplands, a rugged plateau that includes much of the namesake national park.

All three subregions feature a mix of forest and farmland, which the latter prevalent along the route's Warren County course.

[5][6] KY 101 is part of the state primary system from the western junction with US 31W north to its northern terminus.

The highway intersects KY 1533 (Meadow-Port Oliver Road) at Meador before the route crosses the Barren River into Warren County.

[8] KY 101 splits from US 31W onto Chalybeate Road, which heads north and then northeast as the route ascends The Knobs onto a plateau.

The old alignment in that area was redesignated as KY 3611, which became the shortest signed state route in Edmonson County once the designation was assigned in 2013.