I fixed a syntax error, added a leading '>' character to your input string, used perltidy to neaten up the indentation, added the strictures and used the DATA handle to create a self-contained, runnable example. 
use strict;
use warnings;

my %loc;
while (<DATA>) {
    chomp;
    if (/^>(\w+)\s\w+\=(chr\w+)\:(\d+)\-(\d+).*\+(.*)/) {
        $loc{$1} = "$2:$3:$4:$5";
        print $loc{$1} . "\n";
    }
}
__DATA__
>hg19_ensGene_ENST00000237247 range=chr1:67208779-67210057 5'pad=0 3'p
+ad=0 strand=+ repeatMasking=none

[download]
Prints: 
chr1:67208779:67210057: repeatMasking=none

[download]
Also, there is no reason to back-whack = or : or - in your regex. This will also work: 
    if (/^>(\w+)\s\w+=(chr\w+):(\d+)-(\d+).*\+(.*)/) {

[download]

In reply to Re: Using regular expression to extract info from input line by toolic
in thread Using regular expression to extract info from input line by biobee07

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