This new structure was completed in 1603 in record time, as Ieyasu had ordered various daimyō to contribute materials, labor and money for its construction, and the largest three-story yagura was transferred from Gifu Castle to be its tenshu.

Per the 1843 "中山道宿村大概帳" (Nakasendō Shukuson Taigaichō) guidebook issued by the Inspector of Highways (道中奉行, Dōchu-būgyō), the town had a population of 2728 people in 805 houses, including one honjin, one waki-honjin, and 35 hatago.

It extended for approximately 2.3 km (1.4 mi) along the highway,[5] making it the largest post town in Mino Province.

During the Bakumatsu period, Princess Kazu-no-miya stayed at the honjin in Kanō-juku in 1861 en route to marry Shogun Tokugawa Iemochi in Edo.

The 1891 Mino–Owari earthquake and the 1945 Bombing of Gifu in World War II destroyed all of the old buildings at Kanō-juku, with the exception of some structures of the Kanō Tenman-gū, which had originally been built to serve as a place of worship for the castle's residents .