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The film opens with a Spartan elder inspecting a baby which resembles Shrek, he then proceeds to kiss it as the narrator paraphrases the words of Dilios about the Spartan selectiveness of their citizens. The baby vomits on the inspector and is then rejected and punted off the hill. With the next baby in hand, the narrator says if it is Vietnamese, Brangelina gets first dibs. Baby Leonidas is then inspected, having a six-pack, biceps, and beard from birth. He is accepted as a Spartan and prepared for his kinghood through his childhood training, from fighting his grandmother to enduring James Bond-style torture. Leonidas (Sean Maguire) is then cast out into the wild, and survives the harsh winter while killing a giant penguin.

Returning a king for his inauguration ceremony, Leonidas sees Margo (Carmen Electra) dancing and asks her to marry him, to which she responds by giving him the combination to her chastity belt. The film then fast forwards to where the story begins, opening with Leonidas training his son, until Captain (Kevin Sorbo) informs him that a Persian messenger has arrived to present Xerxes' demands for Sparta's submission. Leonidas arrives to greet the messenger in the Spartan way (high-fives for the women and open mouth tongue kisses for the men). After growing angry with the messenger's disrespect, Leonidas kicks him, the messenger's bodyguards, and then several other people he just plain disliked, ranging from Britney Spears (Nicole Parker) to Sanjaya Malakar and the American Idol judges, into "The Pit of Death". As Leonidas walks off he turns to a column, that has a switch that reads "Garbage Disposal", and flips the switch causing the celebrities to spiral to their "death".

Resolving to face the Persians, Leonidas visits the Oracle, proposing an "erotic-sounding" strategy to repel the numerically superior enemy after offering the priests various skin-care lotions for guidance. The Oracle, Ugly Betty, says some well known lines as in "Save the cheerleader, save the world" from Heroes and "Douchebag says what" Leonidas responding with a "What?" Then, the Oracle reveals that Leonidas will die should he go to war. That night, Leonidas and Margo then decide to have sex before Leonidas heads out (which in fact is just Leonidas bench pressing his wife numerous times). Leonidas then goes out to meet the soldiers collected for his departure, and finds that only 13 (not 300) came to the call of war, three among them being Captain, his son (Sonio), and a slightly unfit Spartan named Dilio. The 13 hold hands and skip behind Leonidas to Thermopylae, where they meet Paris Hilton (also played by Parker), whom they rejected from joining their army due to her mild deformities. Leonidas and his platoon soon face off with Xerxes' messenger and his Immortals, beating them in a dance contest before driving them off a cliff. Xerxes (Ken Davitian), impressed, personally approaches Leonidas and attempts to bribe him in a Deal or No Deal fashion. Despite the soldiers encouragements, the Spartan king declines, saying that he will instead make the "God King" fall. The Spartans then face the Xerxes army in a "Yo Momma" fight, ending with a victory in spite of Dilio having his eyes scratched out.

Though victory seemed to be in the Spartans' grasp, Paris Hilton reveals the location of the goat path to Xerxes, having been promised her hump removed among her various demands. Using a CGI army, Xerxes overpowers the 12 remaining Spartans. However, though his army was defeated, Xerxes finds the Transformer cube and transforms a convertible into his suit of armor, playing both the Rick Roll and "Leave Britney Alone" videos to prove his "godly power". But "Xerxestron" ends up delivering on Leonidas's promise to "fall", though he crushed the surviving Spartans under his bulk. The blind Dilio, who left prior to the final battle, eventually returns to Sparta to tell of Leonidas' final moments. A year later, Dilio leads a force of 100 real Spartans and several thousand more CGI ones to defeat the Persians, but the blind warrior ends up going the wrong way and leads the Spartans to Malibu. They end up crashing into Lindsay Lohan while she was coming out of rehab (again), causing her to expose her bare crotch as she flies backwards.

The film ends with a comical musical number set to Gloria Gaynor's I Will Survive performed by all of the characters that have appeared in the film on American Idol, and is followed by additional scenes.