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Biomes are always unlocked after completion
After you've defeated a boss, acquired the key, and entered the next level, here's what happens on a new cycle: Boss fights can be skipped and do not have to be repeated. You can enter the portal to any visited Biome. On the map it's the door with a ring above it. If your weapons are underpowered, you'll be given a Weapon Proficiency boost item at the start of the new Biome.