Place it as the solo reaction beat after the line that causes the confusion, before anyone else responds. Follow it with a confused huh sound button for a two-step reaction, or switch to an oh my god wow button when the moment turns from baffled to shocked.
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This is a three-second clip of a single voice letting out a long, drawn-out 'ah', held longer than a normal sigh or shout. It feels awkward and funny at the same time, the sound of someone realising something too late or having no better reply ready, which makes the pause that follows feel even more uncomfortable and comic.