In that case you could seek to a position close to the end-of-file, walk backward until you find a newline, use a regex to see if the last line matches, and if so, remove it. Then, truncate the file at the current position and append the modified string to it. This would work on a file of any size while using a minimal amount of memory.
Comment on Re^3: Delete last line of file with regex
You've got the right idea but your algorithm is shittily inefficient. The classical (tried and true) solution is to load blocks, one at a time, starting with the last, and search backwards in the block until the marker is found. This solution involves doing some math (to calculate pos from block size and count), but it's much more efficient than using seek.