3
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1 #!/usr/bin/perl
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2
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3 use strict;
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4
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5 use File::Basename;
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6 my $dirname = dirname(__FILE__);
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7
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8 system("wget https://ftp.ncbi.nlm.nih.gov/genomes/GENOME_REPORTS/prokaryotes.txt");
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9 system("wget https://ftp.ncbi.nlm.nih.gov/genomes/GENOME_REPORTS/eukaryotes.txt");
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10
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11 my %continents;
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12 open(F,"countries.txt");
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13 <F>;
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14 while(my $line =<F>){
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15 chomp($line);
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16 my ($continent,$country) = split(/,/,$line);
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17 $continents{$country} = $continent;
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18 }
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19 close(F);
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20
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21 my $input = $ARGV[0];
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22 my $outdir = $ARGV[1];
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23 my $private_genomes = $ARGV[2];
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24
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25 system("cat $input $private_genomes >$outdir/list.txt");
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26
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27 my $concat = "";
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28 open(O2,">$outdir/genbanks.txt");
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29 open(O,">$outdir/strains.txt");
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30 open(GENES,">$outdir/genes.txt");
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31 open(L,">$outdir/list_genomes.txt");
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32 open(L2,">$outdir/list_genomes2.txt");
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33 open(L3,">$outdir/genomes.txt");
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34 open(L4,">$outdir/genomes2.txt");
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35 open(SEQFILE,">$outdir/seqfile");
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36 open(PanSN,">$outdir/all_genomes.fa");
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37 open(METADATA,">$outdir/metadata_strains.txt");
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38
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39 open(F,"$outdir/list.txt");
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40 #open(TEST,">$outdir/test");
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41 while(my $line =<F>){
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42 chomp($line);
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43 my $genbank = $line;
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44 if (!-e "$genbank"){
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45 my $grep = `grep '$line' prokaryotes.txt`;
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46 #print "$genbank $line aaa $grep\n";exit;
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47 my @infos = split(/\t/,$grep);
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48 my $status = $infos[15];
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49 if ($status !~/Complete Genome/ && $status !~/Chromosome/){
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50 #next;
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51 }
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52 my $ftp_path = $infos[$#infos -2];
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53
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54 $ftp_path =~s/ftp:/http:/g;
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55 my @table = split(/\//,$ftp_path);
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56 my $name = $table[$#table];
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57 my $prot_file = "$ftp_path/$name"."_protein.faa.gz";
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58 my $gbff = "$ftp_path/$name"."_genomic.gbff.gz";
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59 my $gff = "$ftp_path/$name"."_genomic.gff.gz";
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60 my $genome_fasta = "$ftp_path/$name"."_genomic.fna.gz";
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61 my @particules = split(/_/,$name);
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62
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63 `wget -O $outdir/$genbank.fasta.gz $genome_fasta`;
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64 `gunzip $outdir/$genbank.fasta.gz`;
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65 `wget -O $outdir/$genbank.gb.gz $gbff`;
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66 system("gunzip $outdir/$genbank.gb.gz");
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67
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68
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69 ################################################################
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70 # for eukaryotes
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71 ################################################################
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72 if (!$grep){
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73 $grep = `grep '$line' eukaryotes.txt`;
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74 my @infos = split(/\t/,$grep);
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75 my $gca = $infos[8];
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76 if ($gca =~/GCA_(\d\d\d)(\d\d\d)(\d\d\d)/){
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77 my $part1 = $1;
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78 $ftp_path = "https://ftp.ncbi.nlm.nih.gov/genomes/all/GCA/$1/$2/$3";
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79 `wget -O $outdir/$gca.index.html $ftp_path`;
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80 my $grep_name = `grep '$gca' $outdir/$gca.index.html`;
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81 unlink("$outdir/$gca.index.html");
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82 my $name;
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83 if ($grep_name =~/href=\"(.*)\"/){
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84 $name = $1;
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85 $name=~s/\///g;
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86 }
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87 $ftp_path = $ftp_path."/$name";
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88 my $prot_file = "$ftp_path/$name"."_protein.faa.gz";
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89 my $gbff = "$ftp_path/$name"."_genomic.gbff.gz";
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90 my $gff = "$ftp_path/$name"."_genomic.gff.gz";
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91 my $genome_fasta = "$ftp_path/$name"."_genomic.fna.gz";
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92 my @particules = split(/_/,$name);
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93
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94 `wget -O $outdir/$genbank.fasta.gz $genome_fasta`;
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95 `gunzip $outdir/$genbank.fasta.gz`;
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96 `wget -O $outdir/$genbank.gb.gz $gbff`;
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97 `wget -O $outdir/$genbank.faa.gz $prot_file`;
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98 system("gunzip $outdir/$genbank.gb.gz");
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99 system("gunzip $outdir/$genbank.faa.gz");
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100
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101 }
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102 }
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103 }
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104 else{
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105 my $genbank_file = $genbank;
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106 my $grep = `grep 'LOCUS' $genbank_file`;
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107 $genbank = "unknown";
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108 if ($grep =~/LOCUS\s+([\-\:\w]+)/){$genbank = $1;}
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109
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110 #$genbank =~s/\:/_/g;
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111
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112 my $cmd = "cp -rf $genbank_file $outdir/$genbank.gb";
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113 system($cmd);
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114
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115 my %genome_seqs;
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116 my $current_chr;
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117 my $go = 0;
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118 open(G,"$outdir/$genbank.gb");
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119 while(<G>){
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120 if ($go == 1 && /(\d+) (.*)$/){
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121 my $line = $2;
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122 $line =~s/ //g;
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123 $genome_seqs{$current_chr}.=$line;
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124 }
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125 if (/LOCUS ([^\s]+)/){
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126 $current_chr = $1;
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127 }
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128 if (/ORIGIN/){$go = 1;}
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129 if (/^\/\//){$go = 0;}
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130 }
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131 close(G);
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132
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133 open(FASTA,">$outdir/$genbank.fasta");
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134 foreach my $ch(keys(%genome_seqs)){
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135 print FASTA ">$ch\n";
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136 my $seq = $genome_seqs{$ch};
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137 print FASTA "$seq\n";
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138 }
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139 close(FASTA);
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140 }
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141 #my $get_organism_line = `head -10 $outdir/$genbank.gb | grep DEFINITION `;
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142 my $get_organism_line = `head -10 $outdir/$genbank.gb | grep -A 1 DEFINITION `;
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143
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144 # if several lines for DEFINITION, concatenate the lines
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145 my @lines_organism = split(/\n/,$get_organism_line);
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146 my $first_line = $lines_organism[0];
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147 my $second_line = $lines_organism[1];
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148 if ($second_line =~/^ (.*)/){
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149 $get_organism_line = $first_line. " ".$1;
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150 }
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151 else{
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152 $get_organism_line = $first_line;
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153 }
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154
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155 my $strain;
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156 my $genus;
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157 if ($get_organism_line =~/DEFINITION (.*)$/){
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158 $strain = $1;
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159 ($genus) = split(/\s/,$strain);
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160 }
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161 my $country = `head -100 $outdir/$genbank.gb | grep country `;
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162 $country =~s/^\s+//g;
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163 $country =~s/\/country=//g;
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164 $country =~s/\"//g;
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165 $country =~s/\n//g;$country =~s/\r//g;
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166 if ($country =~/:/){
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167 my $city;
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168 ($country,$city) = split(/:/,$country);
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169 }
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170 if ($country eq ""){$country = "unresolved";}
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171 my $continent = "unresolved";
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172 if ($continents{$country}){
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173 $continent = $continents{$country};
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174 }
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175 $strain =~s/\.//g;
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176 my ($info1,$info2 ) = split(",",$strain);
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177 $strain = $info1;
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178 $strain =~s/ /_/g;
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179 $strain =~s/strain_//g;
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180 $strain =~s/_chromosome//g;
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181 $strain =~s/_genome//g;
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182 $strain =~s/_str_/_/g;
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183 $strain =~s/[^\w\-\_]//g;
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184 $strain =~s/\-/_/g;
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185
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186 print O "$genbank $strain\n";
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187 $concat .= "$genbank,";
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188 print L "$genbank $outdir/$genbank.gb\n";
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189 print L2 "$genbank\n";
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190 print L3 "$outdir/$genbank.fasta\n";
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191 print L4 "$outdir/$genbank.fasta $strain\n";
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192 print SEQFILE "$genbank $outdir/$genbank.fasta\n";
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193 print METADATA "$strain\t$genus\t$country\t$continent\n";
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194
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195
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196 my %genome_sequences;
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197 my $genome = "";
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198 my $seqid = "";
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199 open(GENOME,"$outdir/$genbank.fasta");
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200 while(<GENOME>){
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201 if (!/^>/){
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202 my $line = $_;
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203 $line =~s/\n//g;$line =~s/\r//g;
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204 $genome_sequences{$seqid} .= $line;
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205 $genome .= $line;
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206 print PanSN $_;
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207 }
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208 elsif (/>([^\s]+)/){
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209 $seqid = $1;
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210 $seqid =~s/\.\d+$//g;
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211 print PanSN ">$strain#$seqid\n";
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212 }
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213 }
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214 close(GENOME);
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215
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216
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217 open(N,">$outdir/$genbank.nuc");
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218 open(P,">$outdir/$genbank.pep");
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219 open(FUNC,">$outdir/$genbank.func");
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220 my $go = 0;
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221 my $start;
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222 my $end;
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223 my $product;
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224 my $complement = 0;
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225 my $end_gene = "no";
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226 my $protein = "";
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227
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228 #`sed -i "s/'//g" $outdir/$genbank.gb`;
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229
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230 my $has_translation = `grep -c 'translation=' $outdir/$genbank.gb`;
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231 $has_translation =~s/\n//g;$has_translation =~s/\r//g;
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232 open(G,"$outdir/$genbank.gb");
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233 my $current_gene;
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234 my $current_chrom;
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235 while(<G>){
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236 if (/^\s+ORGANISM\s+(.*)$/){
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237 }
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238 if (/protein_id=\"(.*)\"/){
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239 $current_gene = $1;
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240 }
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241 if (/LOCUS ([^\s]+)/){
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242 $current_chrom = $1;
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243 }
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244 if (/locus_tag=\"(.*)\"/){
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245 $current_gene = $1;
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246 }
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247 if ($go == 1){
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248 my $line = $_;
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249 $line =~s/ //g;
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250 $line =~s/\n//g;$line =~s/\r//g;
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251 $protein .= $line;
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252 if (/\"$/){
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253 $protein =~s/\"//g;
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254 $end_gene = "yes";
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255
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256 }
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257 }
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258 if (/\/translation=\"(.*)/ or ($has_translation == 0 && /protein_id=/)){
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259 $go = 1;
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260 $protein .= $1;
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261 print P ">$current_gene\n";
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262 print N ">$current_gene\n";
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263 print GENES "$current_gene $product [$strain]\n";
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264
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265 if ($protein =~/\"$/){
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266 $end_gene = "yes";
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267 }
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268 if ($has_translation == 0){$end_gene = "yes";}
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269 }
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270 if ($end_gene eq "yes"){
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271 $protein =~s/\"//g;
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272 print P "$protein\n";
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273 $protein = "";
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274 my $length = $end - $start + 1;
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275
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276 #print "okkkk $current_chrom\n";
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277 #my $geneseq = substr($genome,$start-1,$length);
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278 my $geneseq = substr($genome_sequences{$current_chrom},$start-1,$length);
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279
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280
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281 if ($complement){
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282 my $revcomp = reverse $geneseq;
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283 $revcomp =~ tr/ATGCatgc/TACGtacg/;
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284 $geneseq = $revcomp;
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285 }
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286 print N "$geneseq\n";
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287 print FUNC "$current_gene - $product\n";
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288 $go = 0;
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289 $end_gene = "no";
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290 }
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291 if (/\/product=\"(.*)\"/){
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292 $product = $1;
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293 }
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294 if (/^\s+CDS\s+(\d+)\.\.(\d+)\s*$/){
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295 $start = $1;
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296 $end = $2;
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297 $complement = 0;
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298 }
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299 if (/^\s+CDS\s+complement\((\d+)\.\.(\d+)\)\s*$/){
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300 $start = $1;
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301 $end = $2;
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302 $complement = 1;
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303 }
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304 }
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305 close(G);
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306 close(P);
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307 close(N);
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308 close(FUNC);
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309
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310 if ($has_translation == 0){
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311 system("perl $dirname/translate.pl $outdir/$genbank.nuc $outdir/$genbank.pep");
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312 }
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313
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314 my $prot_num = 0;
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315 open(PRT,">$outdir/$genbank.prt");
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316 open(P,"$outdir/$genbank.pep");
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317 while(<P>){
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318 if (/>(.*)/){
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319 my $prot_id = $1;
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320 $prot_num++;
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321 my $new_id = "$strain"."_".$prot_num;
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322 print PRT ">$new_id\n";
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323 }
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324 else{
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325 print PRT $_;
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326 }
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327 }
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328 close(P);
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329 close(PRT);
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330 }
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331 close(F);
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332 close(O);
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333 close(METADATA);
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334 chop($concat);
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335 print O2 $concat;
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336 close(O2);
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337 close(L);
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338 close(L2);
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339 close(L3);
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340 close(L4);
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341 close(GENES);
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342 close(SEQFILE);
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343 close(PanSN);
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344 #close(TEST);
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345
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346 unlink("prokaryotes.txt");
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347 unlink("eukaryotes.txt");
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