The 301st Fighter Squadron was constituted on 4 July 1942 and assigned to the 332nd Fighter Group at Tuskegee Army Air Field, Alabama, and completed their stateside training at Selfridge Field, Michigan.

In February 1944 the squadron moved to Italy. During its combat tour it was based at Monte Corvino, Capodichino, Ramatelli Airfield, Cattolica, and finally Lucera before returning to the United States in September 1945.

The 301st flew the hottest fighters in the inventory, refining their abilities in the P-40 Tomahawk and P-39 Aircobra from 1943 into 1944, then transitioning to the P-47 Thunderbolt and finishing the war in the P-51 Mustang.

The 301st one of four Army Air Force squadrons known as the Tuskegee Airmen. An all African-American squadron, the 301st's pilots were very determined and capable flyers. Bomber crews were glad to see their red-tailed Mustangs show up for escort duty.

The squadron saw combat in campaigns in the Mediterranean and European Theaters of Operations: Rome-Arno, Normandy, Northern France, Southern France, Northern Apennines, Rhineland, Central Europe, and Po Valley. The squadron was awarded the Distinguished Unit Citation for Combat over Germany on 24 March 1945.

After the war, the squadron was inactivated, re-activated in 1947, and finally disbanded in 1949. The official squadron patch was officially approved on 29 June 1945.