One of the basic questions that people have when they get a kitten (if they didn't purchase it at a pet store or shelter and were told what the sex was) is whether or not it is a boy or a girl! So how do you tell the sex of a kitten?
The easiest thing to do is to try to look for testicles. If you see testicles it is a boy! However if your kitten is a young kitten they may not be noticeable yet. Thus the best way to determine the sex is by measuring the distance between the two holes (anus/vagina or anus/penis).
A female's vagina will be more of a vertical slit and will be fairly close to the anus. A male's penis will be a rounder looking hole and will be farther away from the anus. Thus a male's parts will look like a ':' or colon and be fairly far apart and a female's parts will look more like a ';' or a semicolon.
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Article courtesy of Pet360