On November 25, 2005, at Air Base 5 (BA5), the first squadron of F-16 MLU aircraft in the Portuguese Air Force was formed. The 301 Squadron "Jaguares", which had previously flown the Alpha Jet at Air Base 11 and before that the well-loved "Fiat G-91 Gina", relocated to BA5 and became the first national squadron equipped with F-16 MLU aircraft.

The operation of the F-16 MLU began in February 2003 with the creation of an "F-16 MLU nucleus". This initial group of pilots prepared for the introduction of this new aircraft into the Air Force, ceasing its activities once the new 301 Squadron assumed this responsibility. The Jaguares began this new era with eight pilots and, at the time, three aircraft modified to the MLU version: tail numbers 15133, 15139, and 15121.

Order No. 51/05/A from the Chief of the Air Staff, dated November 11, determined that the new F-16 squadron, now operating with aircraft in the MLU configuration, was assigned the mission of conducting Air Defense and Conventional Attack operations under any weather and lighting conditions.