/usr/bin/seq_cache_populate.pl is in samtools 1.3.1-3.
This file is owned by root:root, with mode 0o755.
The actual contents of the file can be viewed below.
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# The MIT License
# Copyright (c) 2014 Genome Research Ltd.
# Author: Rob Davies <rmd+sam@sanger.ac.uk>
# Permission is hereby granted, free of charge, to any person obtaining a copy
# of this software and associated documentation files (the "Software"), to deal
# in the Software without restriction, including without limitation the rights
# to use, copy, modify, merge, publish, distribute, sublicense, and/or sell
# copies of the Software, and to permit persons to whom the Software is
# furnished to do so, subject to the following conditions:
# The above copyright notice and this permission notice shall be included in
# all copies or substantial portions of the Software.
# THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR
# IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY,
# FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE
# AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER
# LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM,
# OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN
# THE SOFTWARE.
# Import references into a cram reference cache from fasta files.
# See below __END__ for POD documentation.
use strict;
use warnings;
use Digest::MD5;
use Getopt::Long;
use File::Find;
use File::Temp qw(tempfile);
use File::Spec::Functions;
use File::Path 'make_path';
use IO::Handle;
$| = 1;
# Directory where the cache will be built
my $root_dir;
# Number of subdirectories to make below $root_dir
# Each subdir will eat up two hex digits of the file MD5
my $subdirs = 2;
# Directory tree to search when using the -find option
my $find = '';
# How much data to read before spilling to a file
my $max_acc = 256 * 1024 * 1024;
my $usage = "Usage: $0 -root <dir> [-subdirs <n>] input1.fasta ...\n $0 -root <dir> [-subdirs <n>] -find <dir>\n";
# Deal with options
GetOptions("root=s" => \$root_dir, "subdirs=s" => \$subdirs,
"find=s" => \$find) || die $usage;
unless ($root_dir && $subdirs =~ /^\d+$/) { die $usage; }
if ($subdirs >= 16) {
die "$0: Error: -subdirs should be less than 15.\n";
}
# Regexp to convert a hex MD5 to a list of $subdirs subdirectory names, the
# remainder making the filename in the leaf directory
my $dest_regexp = "(..)" x $subdirs . "(" . ".." x (16 - $subdirs) . ")";
my $dest_re = qr/$dest_regexp/;
# Ensure $root_dir exists
unless (-e $root_dir) {
make_path($root_dir);
}
if ($find) {
# Find mode - search a directory tree for anything that looks like a
# fasta file. Any that are found will be put into the new cache, if
# they are not already there.
find({
wanted => sub {
find_files($File::Find::name, $root_dir, $dest_re, $max_acc);
},
no_chdir => 1,
},
$find);
} elsif (@ARGV) {
# If a list of files was given on the command line, go through them
# and try to add each one.
foreach my $name (@ARGV) {
open(my $fh, '<', $name) || die "Couldn't open $name: $!\n";
process_file($name, $fh, $root_dir, $dest_re, $max_acc);
close($fh) || die "Error closing $name: $!\n";
}
} else {
# Otherwise read from STDIN
process_file('STDIN', \*STDIN, $root_dir, $dest_re, $max_acc);
}
exit;
sub find_files {
my ($name, $root_dir, $dest_re, $max_acc) = @_;
# See if $name is a candidate file
my $fh;
return if ($name =~ /~$/); # Ignore backup files
return unless (-f $name && -r _); # Ignore non-regular and unreadable files
# Inspect the first two lines of the candidate
my $buffer;
open($fh, '<', $name) || die "Couldn't open $name: $!\n";
read($fh, $buffer, 8192); # Should be enough to find the header & sequence
close($fh) || die "Error closing $name: $!\n";
my ($l1, $l2) = split(/\n/, $buffer);
# Check for fasta-like content
return unless ($l1 && $l1 =~ /^>\S+/);
return unless ($l2 && $l2 =~ /^[ACGTMRWSYKVHDBNacgtmrwsykvhdbn]+$/);
# Looks like a fasta file, so process it
open($fh, '<', $name) || die "Couldn't open $name: $!\n";
process_file($name, $fh, $root_dir, $dest_re, $max_acc);
close($fh) || die "Error closing $name: $!\n";
}
sub process_file {
my ($name, $in_fh, $root_dir, $dest_re, $max_acc) = @_;
# Process the fasta file $in_fh. Each entry in the file is read, and
# the MD5 calculated as described in the SAM specification (i.e.
# all uppercased with whitespace stripped out). The MD5 is then
# split using $dest_re to convert it into the path name for the entry
# in the cache. If the path is not already present, the entry (in
# uppercased and stripped form) is saved into the cache.
# Entries shorter that $max_acc will be kept in memory. For fasta files
# with lots of short entries this can save a lot of unnecessary writing
# if the data is already in the cache. Anything longer
# gets written out to a file to keep memory consumption under control.
# The temporary files have to be made in $root_dir, as the final
# destination is not known until the entry has been completely read.
my $id; # Name of current fasta entry
my $ctx; # MD5 context
my $acc = ''; # The accumulated sequence
my $tmpfile; # Temporary file name
my $tmpfh; # Temporary file handle
my $extra = 1024; # Extra space to pre-allocate to account for reading
# 1 line past $max_acc
vec($acc, $max_acc + $extra, 8) = 1; # Pre-allocate some space
$acc = '';
# Use an eval block so any stray temporary file can be cleaned up before
# exiting.
eval {
print "Reading $name ...\n";
for (;;) { # Error catching form of while (<>) {...}
undef($!);
last if (eof($in_fh)); # Needed if last line isn't terminated
unless (defined($_ = readline($in_fh))) {
die "Error reading $name: $!" if $!;
last; # EOF
}
if (/^>(\S+)/) {
# Found a fasta header
if ($ctx) { # Finish previous entry, if there is one
finish_entry($id, $ctx, \$acc, $tmpfh, $tmpfile,
$root_dir, $dest_re);
undef($tmpfile);
$acc = '';
}
$id = $1;
$ctx = Digest::MD5->new();
} else {
unless ($id) { die "Found sequence with no header\n"; }
# Read some sequence
chomp;
s/\s+//g;
if ($_) {
$_ = uc($_);
$acc .= $_;
$ctx->add($_);
if (length($acc) > $max_acc) {
# Spill long sequences out to a temporary file in
# $root_dir.
unless ($tmpfile) {
($tmpfh, $tmpfile) = tempfile(DIR => $root_dir,
SUFFIX => '.tmp');
}
print $tmpfh $acc
|| die "Error writing to $tmpfile: $!\n";
$acc = '';
}
}
}
}
if ($ctx) {
# Finish off the last entry
finish_entry($id, $ctx, \$acc, $tmpfh, $tmpfile,
$root_dir, $dest_re);
undef($tmpfile);
}
};
my $err = $@;
if ($tmpfile) { unlink($tmpfile); }
if ($err) { die $err; }
}
sub finish_entry {
my ($id, $ctx, $acc_ref, $tmpfh, $tmpfile, $root_dir, $dest_re) = @_;
# Finish writing an entry
my $digest = $ctx->hexdigest;
# Get the destination directory and filename
my @segs = $digest =~ /$dest_re/;
my $dest_dir = (@segs > 1
? catdir($root_dir, @segs[0..($#segs - 1)])
: $root_dir);
my $dest = catfile($dest_dir, $segs[-1]);
# Make the destination dir if necessary
unless (-e $dest_dir) {
make_path($dest_dir);
}
if (-e $dest) {
# If the file is already present, there's nothing to do apart from
# remove the temporary file if it was made.
print "Already exists: $digest $id\n";
if ($tmpfile) {
close($tmpfh) || die "Error closing $tmpfile: $!\n";
unlink($tmpfile) || die "Couldn't remove $tmpfile: $!\n";
}
} else {
# Need to add the data to the cache.
unless ($tmpfile) {
# If the data hasn't been written already, it needs to be done
# now. Write to a temp file in $dest_dir so if it goes wrong
# we won't leave a file with the right name but half-written
# content.
($tmpfh, $tmpfile) = tempfile(DIR => $dest_dir,
SUFFIX => '.tmp');
}
# Assert that the $tmpfile is now set
unless ($tmpfile) { die "Error: Didn't make a temp file"; }
eval {
# Flush out any remaining data
if ($$acc_ref) {
print $tmpfh $$acc_ref || die "Error writing to $tmpfile: $!\n";
}
# Paranoid file close
$tmpfh->flush() || die "Error flushing to $tmpfile: $!\n";
$tmpfh->sync() || die "Error syncing $tmpfile: $!\n";
close($tmpfh) || die "Error writing to $tmpfile: $!\n";
};
if ($@) {
# Attempt to clean up if writing failed
my $save = $@;
unlink($tmpfile) || warn "Couldn't remove $tmpfile: $!";
die $save;
}
# Finished writing, and everything is flushed as far as possible
# so rename the temp file
print "$dest $id\n";
rename($tmpfile, $dest)
|| die "Error moving $tmpfile to $dest: $!\n";
}
}
__END__
=head1 NAME
seq_cache_populate.pl
=head1 SYNOPSIS
seq_cache_populate.pl -root <dir> [-subdirs <n>] input1.fasta ...
seq_cache_populate.pl -root <dir> [-subdirs <n>] -find <dir>
=head1 DESCRIPTION
Import references into a cram reference cache from fasta files.
When run with a list of fasta files, this program reads the files and stores
the sequences within it in the reference cache under directory <dir>. The
sequences in the cache are stored with names based on the MD5 checksum
of the sequence.
By default, sequences are stored in a hierarchy two directories deep, to
keep the number of items in a single directory to a reasonable number. This
depth can be chaged using the -subdirs option.
If the -find option is used, the program will scan the given directory tree.
Any files that appear to be fasta (by looking for a first line starting with
'>' followed by somthing that looks like DNA sequence) will be read and
added to the reference cache. The traversal will ignore symbolic links.
Samtools/htslib can be made to use the cache by appropriate setting of the
REF_PATH environment varaiable. For example, if seq_cache_populate was run
using options '-root /tmp/ref_cache -subdirs 2', setting REF_PATH to
'/tmp/ref_cache/%2s/%2s/%s' should allow samtools to find the references that
it stored.
Note that if no REF_PATH is specified, htslib will default to downloading from
the EBI reference server and caching locally (see the samtools(1) man page for
details), defaulting to $HOME/.cache/hts-ref/%2s/%2s/%s. This is functionally
equivalent to running this tool with '-root $HOME/.cache/hts-ref -subdirs 2'.
=head1 AUTHOR
Rob Davies.
=cut
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