Those images on top of the buttons show the button mapping directions, all you need to do is click-and-drag from one button to another as needed. Hope this helps!
I’d echo what Dan said (highlight layers are only 2 bit, etc), but as an addition to that, it looks like your highlight layer has two colors in it. That’s a big no-no, as the highlight layer can only display one color at a time.
MPEG-4 is the newest and most efficient when it comes to bitrate vs overall quality so I would recommend that. As for resolutions and framerates that are compliant, it seems like everywhere you look gives you a different answer, but according to the DoStudio manual these are accepted: https://i.imgur.com/cVXAroR.png