Spider-Man: Mystery of the Spider-Woman is a 2007 direct-to-video animated film based on the animated series Spider-Man: The Animated Series, serving as a stand-alone sequel to both Spider-Man: Mask of the Ghost and Spider-Man & the Lizard set on Earth-92131.
A new heroine has arrived in New York whose identity is a mystery—even to Spider-Man. During patrol, the Spider-Man spots her trying to stop one of the Doctor Octopus' shipments on New York's interstate, using a web shooter to send the Doc Ock's truck with its driver off the bridge. Spider-Man saves the driver from falling to his death.
Spider-Man must figure out who the Spider-Woman is and stop two familiar enemies, Doctor Octopus and the Kingpin, from selling illegal weapons to the fictional nation of Symkaria. The two employ Jerry Franklin, a gangster, to provide protection.
Spider-Woman's main focus is on illegal activity by the Doctor Octopus, the Kingpin, and Franklin and she seems uninterested in sparing the lives of her adversaries. Spider-Man sets out to stop the Spider-Woman from making mistakes as she tries to take out the villains and as he encounters numerous twists, setbacks, and apparent false leads in determining her true identity.
Spider-Man suspects one of three women to be Spider-Woman; Jerry Franklin's daughter, Mattie Franklin, a new student, Jessica Drew, who appears to have worked out how to create Spider-Woman's web shooters, and Detective Terri Lee's new partner, Julia Carpenter, who's knowledge of weapons being smudged by Doctor Octopus and Jerry Franklin is much greater than the detective should know.
With Jerry Franklin unable to stop Spider-Woman's raids on the facilities used to hold the various weapons, Doctor Octopus opens a dimensional portal, summoning Venom for additional support to ensure that there are no more losses as a result of the Spider-Woman.
Not long after Venom's arrival in New York, it is revealed that there is not one, but three Spider-Women, all of whom were the women suspected by Spider-Man; Mattie and Julia met taking art classes at college and Julia and Jessica were roommates. They had taken turns to remove suspicion on any one of the three, while using Jessica's technological genius and contempt for the Doctor Octopus (who had framed her father), Mattie's money and access to several key aspects of her father's organisation (Mattie wants to end her father's criminal career, as it led to her mother being killed), and Julia's physical and police skills to ensure that the Kingpin's operation is thwarted (as the crime lord previously left her family in financial ruin after arsonists who worked for him burned down her parents' shop and were not punished due to the lack of sufficient evidence). Carpenter was also saved by Spider-Man three years prior, the event giving the detective the original inspiration for the costumed identity that she now shares with her friends. Jessica and Julia were able to deduce Spider-Man's identity as Peter Parker and how he gained his powers. They and Mattie repeated the experiment, granting themselves similar abilities to him.
In the final confrontation, a ship taking the weapons into international waters for the exchange is destroyed by a bomb planted by Mattie, but not before she is unmasked by Venom. Mattie and Spider-Man narrowly escape the explosion despite the efforts of Venom, who falls into the river and vanishes.
At the conclusion, the GCPD are left to assume that Julia is the only Spider-Woman after she helps rescue Spider-Man from the ship. Julia resigns from the police, due to the potential problems her presence could cause, and decides to leave the city.
Spider-Man gives Julia evidence that he discovered which helps clear Jessica's father. Jerry agrees to testify against the Kingpin and Doctor Octopus after saving Mattie's life during the ship's destruction. Mattie reconciles with her father.
- Christopher Daniel Barnes as Peter Parker/Spider-Man
- TBA as Jessica Drew
- TBA as Julia Carpenter
- TBA as Mattie Franklin
- TBA as Spider-Woman
- Efrem Zimbalist Jr. as Doctor Octopus
- TBA as Jerry Franklin
- Hank Azaria as Venom
- TBA as Jonathan Drew
- Dawnn Lewis as Detective Terri Lee
- Roscoe Lee Brownie as Kingpin
- Julie Bennett as Aunt May