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Building on the literature on agglomeration economies, this study examines how urbanization, industry-diversification, district economics and incubating initiatives are associated to the creation of innovative start-ups in Italy. The empirical analysis is based on a sample of 6018 innovative start-ups distributed across 104 Italian NUTS 3 regions. Our findings show that incubating initiative and industrial districts play a major role for new venture creation and provide support to the positive role of urbanization economies and industry specialization over diversification. Finally, we discuss future research directions grounded on the empirical evidence provided by our study.