During March 1588, Maria d'Aragona, the Marchesa of Vasto, sponsored a set of four intermedi at her palazzo in Chiaia, Naples. The centrepiece of the entertainment was the intermedio entitled ‘Queen Cleopatra on her Ship’. This article explores d'Aragona's role as sponsor of the entertainment, particularly in relation to her interest in the historical figure of Cleopatra. Drawing on sources that informed perceptions of the Egyptian queen during the early- to mid-Cinquecento, it will be shown that within a performance context governed by a strong-willed female patron, the often negatively depicted Cleopatra could be cast as a positive role model, particularly for d'Aragona-related noblewomen who themselves had experienced strong female mentorship and enjoyed the relative autonomy of widowhood. D'Aragona's decision to cast the Neapolitan virtuosa Eufemia Jozola as Cleopatra reinforced the female-orientated nature of the intermedio, and sheds new light on mid-Cinquecento Neapolitan performance practice.