Place it at the end of a sequence showing someone pestering you, after the repeated question has been shown twice, so it works as the payoff. Follow it with Eww Brother Eww to stretch the disgust, or cut to No No Wait Wait if the other person tries to argue back.
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This is a short spoken line, lasting about two seconds, in which a voice dismisses its targets as annoying little flies. The delivery is sharp and fed up, with the insult landing as a quick brush-off rather than a long rant. It feels funny because the complaint is so small and specific, turning a moment of irritation into a playful put-down that works best when someone is being persistently bothersome.