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You can do this with GIFsicle, using the following options: gifsicle -U --disposal=previous --transparent="#ffffff" -O2 anim.gif > anim_trans.gif where anim.gif and anim_trans.gif are the source and destination file names, and #ffffff is the hex code of the color you want to make transparent (here, pure white). (The important options here are -U / ...



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