/usr/bin/blast2sam.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.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 | #!/usr/bin/env perl
#
# Copyright (C) 2009 Genome Research Ltd.
# Portions copyright (C) 2014 Ontario Institute for Cancer Research.
#
# Author: Heng Li <lh3@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.
use strict;
use warnings;
use Getopt::Std;
my $dummy_score;
&blast2sam;
sub blast2sam {
my %opts = ();
getopts('sd', \%opts);
die("Usage: blast2sam.pl <in.blastn>\n") if (-t STDIN && @ARGV == 0);
my ($qlen, $slen, $q, $s, $qbeg, $qend, @sam, @cigar, @cmaux, $show_seq);
$show_seq = defined($opts{s});
$dummy_score = defined($opts{d});
@sam = (); @sam[0,4,6..8,10] = ('', 255, '*', 0, 0, '*');
while (<>) {
if (@cigar && (/^Query=/ || /Score =.*bits.*Expect/ || /^>\S+/)) { # print
&blast_print_sam(\@sam, \@cigar, \@cmaux, $qlen - $qend);
@cigar = ();
}
if (/^Query=\s(\S+)/) {
$sam[2] = undef;
$sam[0] = $1;
my $next_line = <>;
if ($next_line=~/^(\S+)$/) {
$sam[0] .= $1;
}
} elsif (/(\S+)\s+total letters/) {
$qlen = $1; $qlen =~ s/,//g;
} elsif (/^>(\S+)/) {
$sam[2] = $1;
} elsif (/Length\s*=\s*(\d+)/) {
$slen = $1;
} elsif (/Score\s+=\s+(\S+) bits.+Expect(\(\d+\))?\s+=\s+(\S+)/) { # the start of an alignment block
my ($as, $ev) = (int($1 + .499), $3);
$ev = "1$ev" if ($ev =~ /^e/);
@sam[1,3,9,11,12] = (0, 0, '', "AS:i:$as", "EV:Z:$ev");
@cigar = (); $qbeg = 0;
@cmaux = (0, 0, 0, '');
} elsif (/Strand=(\S+)\/(\S+)/) {
$sam[1] |= 0x10 if ($2 eq 'Minus');
} elsif (/Query\s+(\d+)\s*(\S+)\s+(\d+)/) {
$q = $2;
unless ($qbeg) {
$qbeg = $1;
push(@cigar, ($1-1) . "H") if ($1 > 1);
}
$qend = $3;
if ($show_seq) {
my $x = $q;
$x =~ s/-//g; $sam[9] .= $x;
}
} elsif (/Sbjct\:*\s+(\d+)\s*(\S+)\s+(\d+)/) {
$s = $2;
if ($sam[1] & 0x10) {
$sam[3] = $3;
} else {
$sam[3] = $1 unless ($sam[3]);
}
&aln2cm(\@cigar, \$q, \$s, \@cmaux);
}
}
if ($sam[2]) {
&blast_print_sam(\@sam, \@cigar, \@cmaux, $qlen - $qend); # the last argument may be a problem
}
}
sub blast_print_sam {
my ($sam, $cigar, $cmaux, $qrest) = @_;
push(@$cigar, $cmaux->[1] . substr("MDI", $cmaux->[0], 1));
#push(@$cigar, $qrest . 'H') if ($qrest);
if ($sam->[1] & 0x10) {
@$cigar = reverse(@$cigar);
$sam->[9] = reverse($sam->[9]);
$sam->[9] =~ tr/atgcrymkswATGCRYMKSW/tacgyrkmswTACGYRKMSW/;
}
if ($sam->[9]) {
if ($dummy_score) {
$sam->[10] = "";
map {$sam->[10].="I"} (1..length($sam->[9]));
}
} else {
$sam->[9] = '*';
}
$sam->[5] = join('', @$cigar);
print join("\t", @$sam), "\n";
}
sub aln2cm {
my ($cigar, $q, $s, $cmaux) = @_;
my $l = length($$q);
for (my $i = 0; $i < $l; ++$i) {
my $op;
# set $op
if (substr($$q, $i, 1) eq '-') { $op = 1; }
elsif (substr($$s, $i, 1) eq '-') { $op = 2; }
else { $op = 0; }
# for CIGAR
if ($cmaux->[0] == $op) {
++$cmaux->[1];
} else {
push(@$cigar, $cmaux->[1] . substr("MDI", $cmaux->[0], 1));
$cmaux->[0] = $op; $cmaux->[1] = 1;
}
}
}
=head2 SYNOPSIS
blast2sam.pl is a script for parsing output of NCBI's blastn output (default format) into sam format
=over
blast2sam.pl out.blast > out.blast.sam
=back
=head2 OPTIONS
The script has some (hopefully) useful options for tweaking the output sam
B<-s> Print out sequence of the query.
Note that the current implementation prints out
the sequence of aligned query which may be trimmed or otherwise
different from the sequence of raw read in the input fastq. The CIGAR string
is also calculated for this query sequence, not the original read
B<-d> Dummy base quality score will be printed as field #11 in sam file.
Blast output does not have base quality information for a read, so this option
allows to have some fake value instead, may help when using sam file with some
programs. Hardcoded to be a string of 'I' that corresponds to Phred score 40
according to Sanger format.
Using both options:
=over
blast2sam.pl -sd out.blast > out.blast.sam
=back
Note that there is no header generated, so you will need to run
=over
samtools -hT your_ref.fasta your_file.sam > your_file_with_header.sam
=back
=cut
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