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The program runs without any errors, and a seed generates. I thought it was caused by me adding another sprite and enemy, but extracting a whole new, unedited source code folder and running has the same issue.

I tried to run it on a new PC and I got the same error. It is caused by the newest version of Pygame, it seems. Try downgrading to Pygame 2.1.3 and it should work - at least it did for me :-)