Spider-Girl appears in Season 3 of Ultimate Spider-Man. This version is a gender-swapped version of Peter Parker named Petra Parker.
When "Ultimate Spider-Man" arrives in her dimension, he is surprised to find a female version of J. Jonah Jameson named Joanna Jameson, who like Jonah, slanders superheroes. Spidey encounters Spider-Girl, who is very surprised and rather angry that there is a male version of herself, constantly telling Peter to stay out of her way, telling him "a boy can't be Spider-Girl". Peter attempts to warn her of the Green Goblin, who is after her blood. Petra finally relents, having faced a "Goblin" before, and unmasks herself to him, introducing herself, as does he. As they arrive at Oscorp, Spider-Man is surprised to see a female version of Green Goblin, named Norma Osborn (voiced by Wendie Malick), who is based on the original incarnation, with a costume, bat-glider, and pumpkin bombs, while Spider-Girl is surprised to see a male Green Goblin (voiced by Steven Weber, based on the Ultimate incarnation, who is monstrous and mutated, yet still uses a glider and bombs. She questions if "Goblin is a man or monster", which Goblin replies "Why can't I be both?". As Spider-Girl fights Green Goblin, while Spider-Man fights She-Goblin. After finding an opening to steal a sample of Spider-Girl's blood, and after doing so, Norma asks Norman to give her what she asked for, which is a vial of the Goblin serum. After drinking it, Norma transforms into a monstrous version of herself, fully becoming She-Goblin. As they battle and defeat her, Spider-Girl runs out of web-fluid and falls off of the Georgia Washington Bridge, but Peter saves her. Having defeated Norma, Petra thanks Peter for helping her, saying "it's good to know there's another hero out there, even if he's a boy." Spider-Man bids Spider-Girl farewell as he pursues Goblin. Spider-Girl returns in "Spider-Verse" Part 4, having been summoned by the "Ultimate" Spider-Man, alongside Spider-Man 2099, Spyder-Knight, Spider-Man Noir, Spider-Ham and Ultimate Spider-Man, forming the Web-Warriors, in order to combat Goblin, who has injected himself with a serum of their DNA, and has transformed into Spider-Goblin. After defeating Spider-Goblin, Electro, who Goblin had been using as a slave throughout the story, is freed from the Siege Perilous, and having been empowered by it, merges with the Helicarrier, transforming into a massive battle mecha. As they fight Electro, he sets the Helicarrier to self-destruct. After Spider-Man defeats Electro, Spider-Girl, with help from Spider-Ham, set the Helicarrier to fly out of reach, where it can detonate safely. With the battle won, Spider-Girl, along with all of the Spider-Men, returns to her dimension.
- In the comic series Spider-Girl is the daughter of Peter Parker and Mary Jane Watson known as May "Mayday" Parker.
- The scene on the Georgia Washington Bridge is an homage to the comic "The Night Gwen Stacy Died". However here Spider-Man manages to save the girl.