$Path can be different across the two version if the file end with a newline (but yoou say that's ok from command line so..) About this you can replace the loop with my $Path = <$fh>; chomp $Path

Have you tried also C:/Users/DON/Desktop/data.db ?

Anyway the driver part seems to be merged into the path for the db location: the error is for the driver missing.

Have you tried the standard way to connect: my $dbh = DBI->connect("dbi:SQLite:dbname=$dbfile","",""); too? Here the path is a named parameter dbname

You can also workaround (and again does not explain why the packed version fails..) setting both env vars:

$ENV{DBI_DSN} = 'C:/Users/DON/Desktop/data.db' ;
$ENV{DBI_DSNDRIVER} = 'dbi:SQLite';

my $dbh = DBI->connect();
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