Gainesville Lodge No. 41 was Chartered in 1857 by 18 Master Masons who petitioned the Grand Lodge of Florida to form a new Lodge in the City of Gainesville.  The first official meeting was April 15, 1857.  The first new members were brought into the Lodge on May 15, 1857.

For the first 50 years the Lodge met in several different locations in the downtown area.  In 1908 the members of the Lodge purchased a block of property on North Main St. and erected the historic Masonic Lodge Building which still stands.  The dedication ceremony was a major event for the city and thousands of Masons, dignitaries and well-wishers attending the ceremonies.  The building is on the National Historic Register and is an architectural treasure in the city of Gainesville. 

Many distinguished members of the Gainesville Community were members of the Lodge, including Mayors of the City, County and City Commissioners, a President of the University of Florida, Professors and Faculty of the University of Florida, Alachua County Sheriffs, City Police and Fire Officials, many prominent businessmen and distinguished U.S. Military Officers.

In 2020 Gainesville Lodge moved to its new home at 2207 N.W. 13 st.