[1] Mayo's campus is situated on 600 acres located near the intersection of San Pablo Road and J. Turner Butler Boulevard, an expressway which serves as a major thoroughfare to and from the Jacksonville Beaches.

[2][3][4] Impressed by the medical treatment received by members of Jacksonville's Davis family (then owners of the Winn-Dixie supermarket chain) at Mayo Clinic in Minnesota, they rallied community and corporate support to bring Mayo Clinic to Jacksonville and donated a 392-acre parcel off San Pablo Road on which Mayo Clinic opened an outpatient consultation center on October 6th, 1986.

In 2001, after experiencing significant growth in Jacksonville, Mayo Clinic announced its intention to build a hospital on its San Pablo Road campus.

The hospital opened in 2008 and created 304 beds and 22 operating rooms,[1] offering care in more than 35 medical and surgical specialties.

[12] Three additional buildings will be constructed; one will provide integrated services of complex cancers and have space for neurologic and neurosurgical care, one will be a lung restoration center to allow for more transplantations, and the third will include a positron emission tomography (PET) radiochemistry facility with an on-site cyclotron.

[12] In 2019 Mayo announced it will spend $233 million to create a comprehensive cancer center in its Jacksonville campus.

[2] In February 2022 Mayo announced a $432 million project to add five floors to the existing inpatient tower, plus additional facilities at the site.