* Pee-Wee's argument with Francis.
-->'''Pee-Wee:''' I wouldn't sell my bike for all the money in the world. Not for a hundred, million, billion, ''trillion'' dollars!\\
'''Francis:''' Then you're crazy.\\
'''Pee-Wee:''' I know you are but what am I?\\
'''Francis:''' You're a nerd.\\
'''Pee-Wee:''' I know you are but what am I?\\
'''Francis:''' You're an idiot!\\
'''Francis:''' I know you are but what am I?\\
'''Together:''' I know ''you'' are but what am I? [[MotorMouth IKnowYouAreButWhatAmIIKnowYouAreButWhatAmIIKnowYouAreButWhatAmIIKnowYouAreButWhatAmI...]]\\
'''Francis:''' No, I'm not.\\
'''Together:''' You are! No way! Knock it off! Cut it out!\\
'''Francis:''' Oh shut up, Pee-Wee.\\
'''Pee-Wee:''' Why don't you make me?\\
'''Francis:''' Why don't you make ''me''?\\
'''Pee-Wee:''' Because, I don't make monkeys, I just train them.\\
'''Francis:''' Pee-Wee, listen to reason. ''(Pee-Wee put his hand on his ear to "listen")'' Come on!\\
'''Pee-Wee:''' Shhhh, I'm listening to reason.\\
'''Pee-Wee:''' That's my name, don't wear it out.
* Pee-Wee's friend who owns a magic shop pulls out a big head with an EvilLaugh.
-->'''Pee-Wee:''' [[BigNo NOOOOOOOOO!!!]]
* The bath fight with Francis is hilarious.
-->'''Pee-Wee:''' ''(after locking the bathroom door)'' Go ahead; scream your head off! We're ''miles'' from where anyone can hear you! ''(maniacal laughter)''
* Pee-Wee scares some local punks by making an animal sound.
-->'''Punk:''' Hey, man! You're new in this turf. We don't take kindly to strangers around here!\\
'''Pee-Wee:''' GRRROOOWWWWLLL! ''(the gang runs away)''
** Made funnier by the fact that the Punk was played by Tim Burton, the director.
** Still funny later on when Madam Ruby tries to remove his wet jacket and he hisses at her as well, causing her to recoil quickly.
-->'''Madam Ruby:''' Alright, you wanna wear a wet jacket it's alright with Madam Ruby.
* Pee-Wee throws his hitchhiking thumb away when it didn't work.
* The hobo won't stop singing.
* When Pee-Wee wanders the wasteland at night (with just his eyes showing), he breaks out one of his toys: Illuminating goggles. But when he puts them on...
-->'''Aaaaaaaaaaaaaaaaaah!''' [[spoiler: He's surrounded by obviously fake animals.]]
* Pee-wee's impatience in the Alamo tour.
* "I'M TRYING TO USE THE PHONE!"
** Oh my God, just that whole scene at the biker bar. That still makes me laugh every time I see it.
** Pee-Wee cries after he accidentally knocked the motorcycles.
-->"I barely touched them!"
** When [[spoiler:Pee-Wee makes friends with the bikers and they let him borrow a bike. He gets on, rides off...and smashes into a sign.]]
* Pee Wee is on the phone in a public phone booth, and needs to convince the person he's talking to that he's in Texas. So, he loudly sings, "The stars at night, are big and bright--" ... and the ''ENTIRE CROWD'' walking by on the sidewalk stops, turns, claps four times, and sings "Deep in the heaaaaaart of Texas!" in perfect unison.
** Also about Texas, after a rodeo, Pee-wee says he doesn't remember anything...but he remembers the Alamo. Everyone around him cheers.
* The chase through the soundstages and backlots, particularly as "bystanders" get swept up in it.
* "Is this something you can share with the rest of us, Amazing Larry?!"
-->'''Pee-wee:''' Exhibit D: Jimmy what is this? Too late! Chip!
-->'''Chip:''' It looks like a pen.
-->'''Pee-wee:''' Exactly! I bought this pen exactly one hour before my bike was stolen. Why? What's the significance? I DON'T KNOW!
** "Exhibit Q: a scale-model of the entire mall! X marks the scene of the crime. These arrows here show the exact position of the sun at the hour of the crime. Jupiter was aligned with Pluto! The moon was in the seventh..."
* Pee-Wee finally acts against his snake phobia at the burning pet shop.
* The very first scene, as Pee-Wee pulls to the lead in the Tour De France on his coaster bike in his gray suit and tie - sets the mood for the whole movie.
* [[http://www.youtube.com/watch?v=RryZV8NK9-Q And when they finally pulled the driver's body.. from the twisted.. burning.. WRECK, it looked like this!]], doubles as NightmareFuel as well.
* That look Mickey, who just got out of prison, mind you- gives to Pee Wee while he's disguised in drag is both creepy and hilarious at once.
* "Paging Mr. Herman..."
* Right after the car makes a soft landing:
* Pee-wee trying to get into Francis' house.
-->'''Pee-wee:''' I wanna see Francis.\\
'''Butler:''' Francis is busy.\\
'''Pee-wee:''' Busy doing what?\\
'''Butler:''' Busy having his bath.\\
'''Pee-wee:''' Really, where are they hosing him down?\\
''(The Butler slams the door, but Pee-wee knocks again and the Butler answers again)''\\
'''Pee-wee:''' Me again.\\
''(The Butler slams the door again, but hears another knocking sound. He opens the door, but finds a knocking device attached to it, while Pee-wee sneaks by behind him.)''
* The deadly serious reason why Pee-Wee keeps avoiding Dottie's attempts to ask him out to the drive-thru.
** And the CallBack when the same speech [[spoiler:is given by James Brolin.]] Extra funny points for turning [[LameComeback "I know you are, but what am I?"]] into a pick-up line.