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Today, the Negev is home to some 379,000 Jews and some 175,000 Bedouin. At least 80,000 Bedouin citizens live in unrecognized villages under threat of demolition; these citizens are subject to removal at any time via the Removal of Intruders Law.

The region's largest city and administrative capital is Beersheba (pop. 185,000), in the north. At its southern end is the Gulf of Aqaba and the resort city of Eilat. It contains several development towns include Dimona, Arad, Mitzpe Ramon, as well as a number of small Bedouin cities, including Rahat and Tel as-Sabi. There are also several kibbutzim, including Revivim and Sde Boker; the latter became the home of Israel's first Prime Minister, David Ben-Gurion, after his retirement from politics.

The desert is home to the Ben-Gurion University of the Negev, whose faculties include the Jacob Blaustein Institutes for Desert Research and the Albert Katz International School for Desert Studies, both located on the Midreshet Ben-Gurion campus adjacent to Sde Boker.

85% of the Negev is used by the Israel Defense Forces for training purposes. In the remaining portion of the Negev available for civilian purposes, a large number of citizens live together in close proximity to a range of types of hazardous infrastructure, which includes a nuclear reactor, 22 agro and petrochemical factories, an oil terminal, closed military zones, quarries, a toxic waste incinerator Ramat Hovav, cell towers, a power plant, several airports, a prison, and 2 rivers of open sewage