It is located in the Luxton neighbourhood of north-end Winnipeg on Anderson Avenue near Main Street and the Red River.

John West, the first Anglican priest in Western Canada, arrived from England under the auspices of the Church Missionary Society and the Hudson's Bay Company in the Red River Colony (or Selkirk Settlement) to serve as chaplain to the Hudson's Bay officers and men, missionary to the aboriginal people in the area, and pastor to the Selkirk Settlers.

The fourth and present Cathedral was reconstructed in 1926 using most of the stone from the previous building under the guidance and inspiration of Archbishop Samuel P. Matheson.

Archbishop Machray, who succeeded Bishop David Anderson in 1865, is regarded by many local Anglicans as having given unsparingly of his time and talents to St. John's and the Diocese of Rupert's Land for almost forty years.

Since 2002, St. John's Cathedral has been the only Canadian site to host the annual Trinity Conference by web cast from New York.

All the furnishings in the chancel, with the exception of the organ console, can be removed, providing space for concerts, dance recitals, plays and other activities.

Furnishings of interest are the solid brass eagle lectern (known affectionately as Big Bird since it was installed), a gift in 1890 to Archbishop Robert Machray on the 25th anniversary of his consecration as bishop, the intricately carved hexagonal oak pulpit (1907), the Italian marble font (1870), and a Coventry cross, made of nails salvaged from the mediaeval Coventry Cathedral which fell victim to the Blitz in World War II.

James Settee, Archbishop Louis Sherman (bishop), Maj.-Gen. Sir Sam Steele, Archbishop Isaac Stringer, Edna Sutherland, John Sutherland (Canadian Senator), John Barton Taylor, Dr. W. F. Taylor, James Thomson, Lieutenant Governor William Johnston Tupper and Lieutenant Governor Errick Willis.

Among these is the Outreach Ministry which provides financial support in the form of grants to organizations and groups in Winnipeg's North End with programs like literacy projects, community kitchens and bursaries that help to improve and enhance the life of the residents of the area.

Every year the Cathedral congregation contributes to Winnipeg Harvest's Shelf-Help program and also supports the Thelma Wynne Project with clothing for newborns and its own Joshua Tree program which provides children from low-income families with hats, scarves and mittens - most of which are handknit by members of the congregation.