/usr/share/perl5/Bio/Tradis/CommandLine/TradisAnalysis.pm is in bio-tradis 1.3.3+dfsg-3.
This file is owned by root:root, with mode 0o644.
The actual contents of the file can be viewed below.
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# ABSTRACT: Perform full tradis analysis
=head1 SYNOPSIS
Takes a fastq, reference and a tag and generates insertion
site plots for use in Artemis
=cut
use Moose;
use Getopt::Long qw(GetOptionsFromArray);
use Cwd qw(abs_path cwd);
use Bio::Tradis::RunTradis;
use TryCatch;
has 'args' => ( is => 'ro', isa => 'ArrayRef', required => 1 );
has 'script_name' => ( is => 'ro', isa => 'Str', required => 1 );
has 'fastqfile' => ( is => 'rw', isa => 'Str', required => 0 );
has 'tag' => ( is => 'rw', isa => 'Str', required => 0 );
has 'mismatch' => ( is => 'rw', isa => 'Int', required => 0, default => 0 );
has 'tagdirection' =>
( is => 'rw', isa => 'Str', required => 0, default => '3' );
has 'reference' => ( is => 'rw', isa => 'Str', required => 0 );
has 'help' => ( is => 'rw', isa => 'Bool', required => 0 );
has 'mapping_score' =>
( is => 'rw', isa => 'Int', required => 0, default => 30 );
has 'smalt_k' => ( is => 'rw', isa => 'Maybe[Int]', required => 0 );
has 'smalt_s' => ( is => 'rw', isa => 'Maybe[Int]', required => 0 );
has 'smalt_y' => ( is => 'rw', isa => 'Maybe[Num]', required => 0, default => 0.96 );
has 'smalt_r' => ( is => 'rw', isa => 'Maybe[Int]', required => 0, default => -1 );
has 'smalt_n' => ( is => 'rw', isa => 'Maybe[Int]', required => 0, default => 1 );
has 'essentiality' => ( is => 'rw', isa => 'Bool', required => 0, default => 0);
has 'verbose' => ( is => 'rw', isa => 'Bool', default => 0 );
has 'samtools_exec' => ( is => 'rw', isa => 'Str', default => 'samtools' );
has '_output_directory' => (
is => 'rw',
isa => 'Str',
required => 0,
lazy => 1,
builder => '_build__output_directory'
);
has '_stats_handle' => (
is => 'ro',
isa => 'FileHandle',
required => 0,
lazy => 1,
builder => '_build__stats_handle'
);
sub BUILD {
my ($self) = @_;
my (
$fastqfile, $tag, $td, $mismatch, $ref,$smalt_n, $essentiality,
$map_score, $smalt_k, $smalt_s, $smalt_y, $smalt_r, $help, $verbose,$samtools_exec
);
GetOptionsFromArray(
$self->args,
'f|fastqfile=s' => \$fastqfile,
't|tag=s' => \$tag,
'td|tagdirection=i' => \$td,
'mm|mismatch=i' => \$mismatch,
'r|reference=s' => \$ref,
'm|mapping_score=i' => \$map_score,
'sk|smalt_k=i' => \$smalt_k,
'ss|smalt_s=i' => \$smalt_s,
'sy|smalt_y=f' => \$smalt_y,
'n|smalt_n=i' => \$smalt_n,
'sr|smalt_r=i' => \$smalt_r,
'v|verbose' => \$verbose,
'samtools_exec=s' => \$samtools_exec,
'e|essentiality' => \$essentiality,
'h|help' => \$help
);
#Essentiality analysis requires different defaults.
if(!defined($smalt_r) && !defined($map_score) && defined($essentiality))
{
$self->essentiality($essentiality) if ( defined($essentiality));
$smalt_r = 0;
$map_score = 0;
}
$self->fastqfile( abs_path($fastqfile) ) if ( defined($fastqfile) );
$self->tag( uc($tag) ) if ( defined($tag) );
$self->tagdirection($td) if ( defined($td) );
$self->mismatch($mismatch) if ( defined($mismatch) );
$self->reference( abs_path($ref) ) if ( defined($ref) );
$self->mapping_score($map_score) if ( defined($map_score) );
$self->smalt_k($smalt_k) if ( defined($smalt_k) );
$self->smalt_s($smalt_s) if ( defined($smalt_s) );
$self->smalt_y($smalt_y) if ( defined($smalt_y) );
$self->smalt_r($smalt_r) if ( defined($smalt_r) );
$self->smalt_n($smalt_n) if ( defined($smalt_n) );
$self->help($help) if ( defined($help) );
$self->verbose($verbose) if ( defined($verbose));
$self->samtools_exec($samtools_exec) if ( defined($samtools_exec) );
# print usage text if required parameters are not present
( $fastqfile && $tag && $ref ) or die $self->usage_text;
}
sub run {
my ($self) = @_;
if ( defined( $self->help ) ) {
#if ( scalar( @{ $self->args } ) == 0 ) {
$self->usage_text;
}
#parse list of files and run pipeline for each one if they all exist
my $fq = $self->fastqfile;
open( FILES, "<", $fq ) or die "Cannot find $fq";
my @filelist = <FILES>;
my $file_dir = $self->get_file_dir;
#check files exist before running
my $line_no = 0;
my $full_path;
foreach my $f1 (@filelist) {
chomp($f1);
$line_no++;
if ( $f1 =~ /^\// ) { $full_path = $f1; }
else { $full_path = "$file_dir/$f1"; }
unless ( -e $full_path ) {
die "File $full_path does not exist ($fq, line $line_no)\n";
}
}
#if all files exist, continue with analysis
my $at_least_one_good_fastq = 0;
foreach my $f2 (@filelist) {
chomp($f2);
if ( $f2 =~ /^\// ) { $full_path = $f2; }
else { $full_path = "$file_dir/$f2"; }
my $analysis = Bio::Tradis::RunTradis->new(
fastqfile => $full_path,
tag => $self->tag,
tagdirection => $self->tagdirection,
mismatch => $self->mismatch,
reference => $self->reference,
mapping_score => $self->mapping_score,
output_directory => $self->_output_directory,
_stats_handle => $self->_stats_handle,
smalt_k => $self->smalt_k,
smalt_s => $self->smalt_s,
smalt_y => $self->smalt_y,
smalt_r => $self->smalt_r,
smalt_n => $self->smalt_n,
verbose => $self->verbose,
samtools_exec => $self->samtools_exec
);
try {
$analysis->run_tradis;
$at_least_one_good_fastq = 1;
}
catch (Bio::Tradis::Exception::TagFilterError $e) {
my $tag = $self->tag;
warn "There was a problem filtering '$full_path' by '$tag'; it looks like the tag was not found in any read\n";
}
}
if ( ! $at_least_one_good_fastq ) {
Bio::Tradis::Exception::TagFilterError->throw( error => "None of the input files contained the specified tag. Please check that your inputs are valid fastq files and that at least one read in one of them starts with the specified tag\n" );
}
$self->_tidy_stats;
close(FILES);
#$self->_combine_plots;
}
sub _build__output_directory {
return cwd();
}
sub _stats_filename {
my ($self) = @_;
my $dir = $self->get_file_dir;
my $output_directory = $self->_output_directory;
my $stats_filename = $self->fastqfile;
$stats_filename =~ s/$dir\///;
$stats_filename =~ s/[^\.]+$/stats/;
return "$output_directory/$stats_filename";
}
sub _build__stats_handle {
my ($self) = @_;
my $stats_filename = $self->_stats_filename();
open( my $stats, ">", $stats_filename );
return $stats;
}
sub _tidy_stats {
my ($self) = @_;
my $output_directory = $self->_output_directory;
my $stats_filename = $self->_stats_filename();
open( STATS, '<', $stats_filename );
open( TMP, '>', "$output_directory/tmp.stats" );
my $header = 0;
while ( my $line = <STATS> ) {
if ( $line =~ /^File/ ) {
if ( $header == 0 ) {
print TMP "$line";
$header = 1;
}
}
else {
print TMP "$line";
}
}
close(TMP);
close(STATS);
system("mv $output_directory/tmp.stats $stats_filename");
}
sub _combine_plots {
my ($self) = @_;
my $filelist = $self->fastqfile;
return 1;
}
sub get_file_dir {
my ($self) = @_;
my $fq = $self->fastqfile;
my @dirs = split( '/', $fq );
pop(@dirs);
return join( '/', @dirs );
}
sub usage_text {
print <<USAGE;
Run a TraDIS analysis. This involves:
1: filtering the data with tags matching that passed via -t option
2: removing the tags from the sequences
3: mapping
4: creating an insertion site plot
5: creating a stats summary
Usage: bacteria_tradis [options]
Options:
-f : text file listing fastq files with tradis tags attached
-t : tag to search for
-r : reference genome in fasta format (.fa)
-td : tag direction - 3 or 5 (optional. default = 3)
-mm : number of mismatches allowed when matching tag (optional. default = 0)
-m : mapping quality cutoff score (optional. default = 30)
--smalt_k : custom k-mer value for SMALT mapping (optional)
--smalt_s : custom step size for SMALT mapping (optional)
--smalt_y : custom y parameter for SMALT (optional. default = 0.96)
--smalt_r : custom r parameter for SMALT (optional. default = -1)
-n : number of threads to use for SMALT and samtools sort (optional. default = 1)
-e : set defaults for essentiality experiment (smalt_r = 0, -m = 0)
-v : verbose debugging output
USAGE
exit;
}
__PACKAGE__->meta->make_immutable;
no Moose;
1;
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