The purpose of this study was to analyze the spatial distribution and location factors of startups by generation (young and middle-aged) in Seoul. To this end, a research model was established that included factors of industry, population, and startup institutions by generation in 424 administrative districts using the Seoul Business Enterprise Survey(2018), which includes data on the age group of entrepreneurs. As an analysis method, descriptive statistics were conducted to confirm the frequency, average and standard deviation of startups by generation and major variables in the administrative districts of Seoul, and spatial distribution and characteristics of startups by generation were analyzed through global and local spatial autocorrelation analysis. In particular, the spatial distribution of startups in Seoul was confirmed in-depth by categorizing and analyzing startups by major industries. Afterwards, an appropriate spatial regression analysis model was selected through the Lagrange test, and based on this, the location factors affecting startups by generation were analyzed. The main results derived from the research results are as follows. First, there was a significant difference in the spatial distribution of young and middle-aged startups. The young people started to startups in the belt-shaped area that connects Seocho·Gangnam-Yongsan-Mapo-Gangseo, while middle-aged people were relatively active in the southeastern region represented by Seocho, Gangnam, Songpa, and Gangdong. Second, startups by generation in Seoul showed various spatial distributions according to the type of business. In the knowledge high-tech industries(ICT, professional services) in common, Seocho, Gangnam, Mapo, Guro, and Geumcheon were the centers, and the manufacturing industry was focused on existing clusters. On the other hand, in the case of the life service industry, young people were active in startups near universities and cultural centers, while middle-aged people were concentrated on new towns. Third, there was a difference in factors that influenced the startup location of each generation in Seoul. For young people, high-tech industries, universities, cultural capital, and densely populated areas were significant factors for startup, and for middle-aged people, professional service areas, low average age, and the level of concentration of start-up support institutions had a significant influence on startup. Also, these location factors had different influences for each industry. The implications suggested through the study are as follows. First, it is necessary to support systematic startups considering the characteristics of each region, industry, and generation in Seoul. As there are significant differences in startup regions and industries by generation, it is necessary to strengthen a customized startup support system that takes into account these regional and industrial characteristics. Second, in terms of research methods, a follow-up study is needed that comprehensively considers culture and finance at the large districts(Gu) level through data accumulation.