#!/usr/bin/env perl #------------------------------------------------------------------------------- # Programmer: Adam M Phillippy, The Institute for Genomic Research # File: nucmer # Date: 04 / 09 / 03 # # Usage: # nucmer [options] # # Try 'nucmer -h' for more information. # # Purpose: To create alignments between two multi-FASTA inputs by using # the MUMmer matching and clustering algorithms. # #------------------------------------------------------------------------------- use lib "./ext/mummer/lintel/scripts"; use Foundation; use File::Spec::Functions; use strict; my $AUX_BIN_DIR = "./ext/mummer/lintel/aux_bin"; my $BIN_DIR = "./ext/mummer/lintel"; my $SCRIPT_DIR = "./ext/mummer/lintel/scripts"; my $VERSION_INFO = q~ NUCmer (NUCleotide MUMmer) version 3.06 ~; my $HELP_INFO = q~ USAGE: nucmer [options] DESCRIPTION: nucmer generates nucleotide alignments between two mutli-FASTA input files. Two output files are generated. The .cluster output file lists clusters of matches between each sequence. The .delta file lists the distance between insertions and deletions that produce maximal scoring alignments between each sequence. MANDATORY: Reference Set the input reference multi-FASTA filename Query Set the input query multi-FASTA filename OPTIONS: --mum Use anchor matches that are unique in both the reference and query --mumcand Same as --mumreference --mumreference Use anchor matches that are unique in in the reference but not necessarily unique in the query (default behavior) --maxmatch Use all anchor matches regardless of their uniqueness -b|breaklen Set the distance an alignment extension will attempt to extend poor scoring regions before giving up (default 200) -c|mincluster Sets the minimum length of a cluster of matches (default 65) --[no]delta Toggle the creation of the delta file (default --delta) --depend Print the dependency information and exit -d|diagfactor Set the clustering diagonal difference separation factor (default 0.12) --[no]extend Toggle the cluster extension step (default --extend) -f --forward Use only the forward strand of the Query sequences -g|maxgap Set the maximum gap between two adjacent matches in a cluster (default 90) -h --help Display help information and exit -l|minmatch Set the minimum length of a single match (default 20) -o --coords Automatically generate the original NUCmer1.1 coords output file using the 'show-coords' program --[no]optimize Toggle alignment score optimization, i.e. if an alignment extension reaches the end of a sequence, it will backtrack to optimize the alignment score instead of terminating the alignment at the end of the sequence (default --optimize) -p|prefix Set the prefix of the output files (default "out") -r --reverse Use only the reverse complement of the Query sequences --[no]simplify Simplify alignments by removing shadowed clusters. Turn this option off if aligning a sequence to itself to look for repeats (default --simplify) -V --version Display the version information and exit ~; my $USAGE_INFO = q~ USAGE: nucmer [options] ~; my @DEPEND_INFO = ( "$BIN_DIR/mummer", "$BIN_DIR/mgaps", "$BIN_DIR/show-coords", "$AUX_BIN_DIR/postnuc", "$AUX_BIN_DIR/prenuc", "$SCRIPT_DIR/Foundation.pm" ); my %DEFAULT_PARAMETERS = ( "OUTPUT_PREFIX" => "out", # prefix for all output files "MATCH_ALGORITHM" => "-mumreference", # match finding algo switch "MATCH_DIRECTION" => "-b", # match direction switch "MIN_MATCH" => "20", # minimum match size "MAX_GAP" => "90", # maximum gap between matches "MIN_CLUSTER" => "65", # minimum cluster size "DIAG_FACTOR" => ".12", # diagonal difference fraction "BREAK_LEN" => "200", # extension break length "POST_SWITCHES" => "" # switches for the post processing ); sub main ( ) { my $tigr; # TIGR::Foundation object my @err; # Error variable my $ref_file; # path of the reference input file my $qry_file; # path of the query input file #-- The command line options for the various programs my $pfx = $DEFAULT_PARAMETERS { "OUTPUT_PREFIX" }; my $algo = $DEFAULT_PARAMETERS { "MATCH_ALGORITHM" }; my $mdir = $DEFAULT_PARAMETERS { "MATCH_DIRECTION" }; my $size = $DEFAULT_PARAMETERS { "MIN_MATCH" }; my $gap = $DEFAULT_PARAMETERS { "MAX_GAP" }; my $clus = $DEFAULT_PARAMETERS { "MIN_CLUSTER" }; my $diff = $DEFAULT_PARAMETERS { "DIAG_FACTOR" }; my $blen = $DEFAULT_PARAMETERS { "BREAK_LEN" }; my $psw = $DEFAULT_PARAMETERS { "POST_SWITCHES" }; my $fwd; # if true, use forward strand my $rev; # if true, use reverse strand my $maxmatch; # matching algorithm switches my $mumreference; my $mum; my $extend = 1; # if true, extend clusters my $delta = 1; # if true, create the delta file my $optimize = 1; # if true, optimize alignment scores my $simplify = 1; # if true, simplify shadowed alignments my $generate_coords; #-- Initialize TIGR::Foundation $tigr = new TIGR::Foundation; if ( !defined ($tigr) ) { print (STDERR "ERROR: TIGR::Foundation could not be initialized"); exit (1); } #-- Set help and usage information $tigr->setHelpInfo ($HELP_INFO); $tigr->setUsageInfo ($USAGE_INFO); $tigr->setVersionInfo ($VERSION_INFO); $tigr->addDependInfo (@DEPEND_INFO); #-- Get command line parameters $err[0] = $tigr->TIGR_GetOptions ( "maxmatch" => \$maxmatch, "mumcand" => \$mumreference, "mumreference" => \$mumreference, "mum" => \$mum, "b|breaklen=i" => \$blen, "c|mincluster=i" => \$clus, "delta!" => \$delta, "d|diagfactor=f" => \$diff, "extend!" => \$extend, "f|forward" => \$fwd, "g|maxgap=i" => \$gap, "l|minmatch=i" => \$size, "o|coords" => \$generate_coords, "optimize!" => \$optimize, "p|prefix=s" => \$pfx, "r|reverse" => \$rev, "simplify!" => \$simplify ); #-- Check if the parsing was successful if ( $err[0] == 0 || $#ARGV != 1 ) { $tigr->printUsageInfo( ); print (STDERR "Try '$0 -h' for more information.\n"); exit (1); } $ref_file = File::Spec->rel2abs ($ARGV[0]); $qry_file = File::Spec->rel2abs ($ARGV[1]); #-- Set up the program parameters if ( $fwd && $rev ) { $mdir = "-b"; } elsif ( $fwd ) { $mdir = ""; } elsif ( $rev ) { $mdir = "-r"; } if ( ! $extend ) { $psw .= "-e "; } if ( ! $delta ) { $psw .= "-d "; } if ( ! $optimize ) { $psw .= "-t "; } if ( ! $simplify ) { $psw .= "-s "; } undef (@err); $err[0] = 0; if ( $mum ) { $err[0] ++; $algo = "-mum"; } if ( $mumreference ) { $err[0] ++; $algo = "-mumreference"; } if ( $maxmatch ) { $err[0] ++; $algo = "-maxmatch"; } if ( $err[0] > 1 ) { $tigr->printUsageInfo( ); print (STDERR "ERROR: Multiple matching algorithms selected\n"); print (STDERR "Try '$0 -h' for more information.\n"); exit (1); } #-- Set up the program path names my $algo_path = "$BIN_DIR/mummer"; my $mgaps_path = "$BIN_DIR/mgaps"; my $prenuc_path = "$AUX_BIN_DIR/prenuc"; my $postnuc_path = "$AUX_BIN_DIR/postnuc"; my $showcoords_path = "$BIN_DIR/show-coords"; #-- Check that the files needed are all there and readable/writable { undef (@err); if ( !$tigr->isExecutableFile ($algo_path) ) { push (@err, $algo_path); } if ( !$tigr->isExecutableFile ($mgaps_path) ) { push (@err, $mgaps_path); } if ( !$tigr->isExecutableFile ($prenuc_path) ) { push (@err, $prenuc_path); } if ( !$tigr->isExecutableFile ($postnuc_path) ) { push (@err, $postnuc_path); } if ( !$tigr->isReadableFile ($ref_file) ) { push (@err, $ref_file); } if ( !$tigr->isReadableFile ($qry_file) ) { push (@err, $qry_file); } if ( !$tigr->isCreatableFile ("$pfx.ntref") ) { if ( !$tigr->isWritableFile ("$pfx.ntref") ) { push (@err, "$pfx.ntref"); } } if ( !$tigr->isCreatableFile ("$pfx.mgaps") ) { if ( !$tigr->isWritableFile ("$pfx.mgaps") ) { push (@err, "$pfx.mgaps"); } } if ( !$tigr->isCreatableFile ("$pfx.cluster") ) { if ( !$tigr->isWritableFile ("$pfx.cluster") ) { push (@err, "$pfx.cluster"); } } if ( !$tigr->isCreatableFile ("$pfx.delta") ) { if ( !$tigr->isWritableFile ("$pfx.delta") ) { push (@err, "$pfx.delta"); } } if ( $generate_coords ) { if ( !$tigr->isExecutableFile ($showcoords_path) ) { push (@err, $showcoords_path); } if ( !$tigr->isCreatableFile ("$pfx.coords") ) { if ( !$tigr->isWritableFile ("$pfx.coords") ) { push (@err, "$pfx.coords"); } } } #-- If 1 or more files could not be processed, terminate script if ( $#err >= 0 ) { $tigr->logError ("ERROR: The following critical files could not be used", 1); while ( $#err >= 0 ) { $tigr->logError (pop(@err), 1); } $tigr->logError ("Check your paths and file permissions and try again", 1); $tigr->bail( ); } } #-- Run prenuc and assert return value is zero print (STDERR "1: PREPARING DATA\n"); $err[0] = $tigr->runCommand ("$prenuc_path $ref_file > $pfx.ntref"); if ( $err[0] != 0 ) { $tigr->bail ("ERROR: prenuc returned non-zero, please file a bug report\n"); } #-- Run mummer | mgaps and assert return value is zero print (STDERR "2,3: RUNNING mummer AND CREATING CLUSTERS\n"); open(ALGO_PIPE, "$algo_path $algo $mdir -l $size -n $pfx.ntref $qry_file |") or $tigr->bail ("ERROR: could not open $algo_path output pipe $!"); open(CLUS_PIPE, "| $mgaps_path -l $clus -s $gap -f $diff > $pfx.mgaps") or $tigr->bail ("ERROR: could not open $mgaps_path input pipe $!"); while ( ) { print CLUS_PIPE or $tigr->bail ("ERROR: could not write to $mgaps_path pipe $!"); } $err[0] = close(ALGO_PIPE); $err[1] = close(CLUS_PIPE); if ( $err[0] == 0 || $err[1] == 0 ) { $tigr->bail ("ERROR: mummer and/or mgaps returned non-zero,". " please file a bug report\n"); } #-- Run postnuc and assert return value is zero print (STDERR "4: FINISHING DATA\n"); $err[0] = $tigr->runCommand ("$postnuc_path $psw -b $blen $ref_file $qry_file $pfx < $pfx.mgaps"); if ( $err[0] != 0 ) { $tigr->bail ("ERROR: postnuc returned non-zero,". " please file a bug report\n"); } #-- If the -o flag was set, run show-coords using NUCmer1.1 settings if ( $generate_coords ) { print (STDERR "5: GENERATING COORDS FILE\n"); $err[0] = $tigr->runCommand ("$showcoords_path -r $pfx.delta > $pfx.coords"); if ( $err[0] != 0 ) { $tigr->bail ("ERROR: show-coords returned non-zero,". " please file a bug report\n"); } } #-- Remove the temporary output $err[0] = unlink ("$pfx.ntref", "$pfx.mgaps"); if ( $err[0] != 2 ) { $tigr->logError ("WARNING: there was a problem deleting". " the temporary output files", 1); } #-- Return success return (0); } exit ( main ( ) ); #-- END OF SCRIPT