in reply to Re: =~ matches non-existent symbols
in thread =~ matches non-existent symbols

Check your file with
perl -mcharnames -e '<> =~ m/([^atcg])/i; print charnames::viacode(ord + $1), "\n"' ATCG_FILE
I created one file in Vim, typed 'atcgATCG' there, saved it, run the one-liner and got:
LINE FEED
Many editors add newline to the end of a file when they save it.