Place it at the peak of a mishap, after a Dramatic Dun Dun Dun build-up and just before the consequences land, then follow it with SpongeBob Fail to underline the flop. Keep it as the main reaction in the sequence so the surrounding gags support it rather than compete with it.
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This is a single wordless scream held high for about two and a half seconds, with a strained, panicked voice and nothing else layered underneath. It hits as pure sudden fear, the sound of realising something has just gone badly wrong, and it works because listeners recognise that panic instantly and brace for a fail, a scare or an over-the-top reaction to follow.