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author | cpt |
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date | Mon, 05 Jun 2023 02:47:24 +0000 |
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package CPT::Bio::ORF; use strict; use warnings; use autodie; use Moose; has min_gene_length => ( is => 'rw', isa => 'Int', default => sub { 0 }, ); has sc_atg => ( is => 'rw', isa => 'Bool', default => sub { 1 } ); has sc_ttg => ( is => 'rw', isa => 'Bool', default => sub { 1 } ); has sc_ctg => ( is => 'rw', isa => 'Bool', default => sub { 0 } ); has sc_gtg => ( is => 'rw', isa => 'Bool', default => sub { 1 } ); our %code = ( "TTT" => "F", "TTC" => "F", "TTA" => "L", "TTG" => "L", "TCT" => "S", "TCC" => "S", "TCA" => "S", "TCG" => "S", "TAT" => "Y", "TAC" => "Y", "TAA" => "*", "TAG" => "*", "TGT" => "C", "TGC" => "C", "TGA" => "*", "TGG" => "W", "CTT" => "L", "CTC" => "L", "CTA" => "L", "CTG" => "L", "CCT" => "P", "CCC" => "P", "CCA" => "P", "CCG" => "P", "CAT" => "H", "CAC" => "H", "CAA" => "Q", "CAG" => "Q", "CGT" => "R", "CGC" => "R", "CGA" => "R", "CGG" => "R", "ATT" => "I", "ATC" => "I", "ATA" => "I", "ATG" => "M", "ACT" => "T", "ACC" => "T", "ACA" => "T", "ACG" => "T", "AAT" => "N", "AAC" => "N", "AAA" => "K", "AAG" => "K", "AGT" => "S", "AGC" => "S", "AGA" => "R", "AGG" => "R", "GTT" => "V", "GTC" => "V", "GTA" => "V", "GTG" => "V", "GCT" => "A", "GCC" => "A", "GCA" => "A", "GCG" => "A", "GAT" => "D", "GAC" => "D", "GAA" => "E", "GAG" => "E", "GGT" => "G", "GGC" => "G", "GGA" => "G", "GGG" => "G", ); sub run { my ($self, $sequence) = @_; # Read through forward strand my @putative_starts; # 30 seconds with a bioperl object # 5 seconds with string munging. >:| my $dna = uc( $sequence ); my $length = length($sequence); # Pre-create the regular expressions my ( $regex_forward, $regex_backwards ); my $not_statement_f = '^'; my $not_statement_r = '^'; if ( !$self->sc_atg() ) { $not_statement_f .= 'A'; $not_statement_r .= 'T'; } if ( !$self->sc_ctg() ) { $not_statement_f .= 'C'; $not_statement_r .= 'G'; } if ( !$self->sc_ttg() ) { $not_statement_f .= 'T'; $not_statement_r .= 'A'; } if ( !$self->sc_gtg() ) { $not_statement_f .= 'G'; $not_statement_r .= 'C'; } # If any start is acceptable, we re-add them and remove our ^ if($not_statement_r eq '^' && $not_statement_f eq '^'){ $not_statement_f = 'ACTG'; $not_statement_r = 'ACTG'; } $regex_forward = qr/[${not_statement_f}]TG/; $regex_backwards = qr/CA[${not_statement_r}]/; # Collect putative starts for ( my $i = 1 ; $i < $length - 1 ; $i++ ) { my $tri_nt = substr( $dna, $i - 1, 3 ); #$seq_obj->subseq($i,$i+2); if ( $tri_nt =~ $regex_forward ) { push( @putative_starts, [ $i, '+' ] ); } if ( $tri_nt =~ $regex_backwards ) { push( @putative_starts, [ $i + 2, '-' ] ); } } my %ORFs; #Loop through all of the starts we have my $fc = 0; my $rc = 0; foreach (@putative_starts) { my @putative_start = @{$_}; my $final_seq = ""; my $add; my $tri_nt; if ( $putative_start[1] eq "+" ) { my $end; for ( my $k = $putative_start[0] ; $k < $length ; $k = $k + 3 ) { my $tri_nt = substr( $dna, $k, 3 ); my $aa = $code{$tri_nt}; if ( $aa && $aa ne '*' ) { $end = $k + 3; $final_seq .= $tri_nt; } else { last; } } if ( length($final_seq)/3 > $self->min_gene_length() ) { $ORFs{ 'f_' + $fc++ } = [ length($final_seq)/3, $putative_start[0], $end, 'F', $final_seq ]; } } # - strand else { my $end; for ( my $k = $putative_start[0] ; $k >= 2 ; $k = $k - 3 ) { my $tmp = reverse( substr( $dna, $k - 3, 3 ) ); $tmp =~ tr/ACTG/qzAC/; $tmp =~ tr/qz/TG/; my $aa = $code{$tmp}; if ( defined $aa && $aa ne '*' ) { $end = $k - 1; $final_seq .= $tmp; } else { last; } } if ( length($final_seq)/3 > $self->min_gene_length() ) { $ORFs{ 'r_' + $rc++ } = [ length($final_seq)/3, $end, $putative_start[0], 'R', $final_seq ]; } } } my @orfs; for my $orf_key ( sort( keys(%ORFs) ) ) { my @tmp= @{ $ORFs{$orf_key} }; my $seqobj = Bio::Seq->new( -display_id => sprintf( 'orf%05d_%s', ($orf_key + 1), $tmp[3], ), -desc => sprintf( '[%s-%s; %s aa long]' ,$tmp[1], $tmp[2], $tmp[0] ), -seq => $tmp[4] ); push(@orfs, $seqobj); } return @orfs; } no Moose; 1; __END__ =pod =encoding UTF-8 =head1 NAME CPT::Bio::ORF =head1 VERSION version 1.99.4 =function run =head1 AUTHOR Eric Rasche <rasche.eric@yandex.ru> =head1 COPYRIGHT AND LICENSE This software is Copyright (c) 2014 by Eric Rasche. This is free software, licensed under: The GNU General Public License, Version 3, June 2007 =cut