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After receiving a mysterious signal from an unknown base, Spider-Man traces it to an abandoned building in the hopes of finding Nick Fury.
Doc Ock and his new assistant, Morbius, create Anti-Venom in order to destroy one of the Web Warriors.
Spider-Man faces the culmination of all Doc Ock's evil machinations when the new Sinister Six assaults the Web Warriors and S.H.I.E.L.D. Academy at the Triskelion.
As Season 4 begins, Doctor Octopus allies himself with Arnim Zola and HYDRA and begins to form a new iteration of the Sinister Six starting with Kraven the Hunter. When Doctor Octopus attempts to turn Norman Osborn back into Green Goblin, he finds that Osborn injected himself with an anti-Goblin serum to prevent further transformations. Doctor Octopus uses the Siege Perilous to summon the Green Goblin's demonic winged counterpart from Miles Morales's universe and after a brief battle recruits him instead. Then it is revealed that Rhino has become a spy for Doctor Octopus while operating as a student at the S.H.I.E.L.D. Academy in exchange for finding a cure for his condition (since he gained his powers by drinking one of Doc Ock's animal serums). Now titled Ultimate Spider-Man vs the Sinister 6, the show will see Spider-Man fighting evil with the help of S.H.I.E.L.D. and his own team consisting of Agent Venom, Iron Spider, Miles Morales (going by the name Kid Arachnid), and the newly introduced Kaine/Scarlet Spider, as well as S.H.I.E.L.D. operatives Leo Fitz and Jemma Simmons.