Class: BacterialComparator

Inherits:
Object
  • Object
show all
Defined in:
lib/bacterial-comparator.rb

Instance Attribute Summary collapse

Instance Method Summary collapse

Constructor Details

#initialize(options, root) ⇒ BacterialComparator

Initialize BacterialAnnotator options, options, ROOT, options, options)



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# File 'lib/bacterial-comparator.rb', line 22

def initialize options, root

  @root = root
  @outdir = File.expand_path(File.dirname(options[:outdir])) + "/#{File.basename(options[:outdir])}"
  Dir.mkdir(@outdir) if ! Dir.exists? @outdir
  @genomes_list = options[:genomes_list]
  @proc = options[:proc].to_i
  @phylo_nb_genes = options[:phylo_nb_genes]
  @refgenome_file = options[:refgenome]
  @refgenome = ""

  if ["fasttree","raxml"].include? options[:software]
    @software = options[:software]
  else
    @software = "fasttree"
  end

  min_cov = options[:min_cov].to_f
  min_pid = options[:pidentity].to_f
  if min_cov > 1
    min_cov = min_cov/100
  end
  if min_pid > 1
    min_pid = min_pid/100
  end

  @aln_opt = options[:align].downcase
  @run_phylo = 0
  if options[:phylogeny] == 1
    @bootstrap = options[:bootstrap]
    @run_phylo = 1
  end

  @ref_prot = get_ref_prot
  @synteny = read_prot_synteny
  @stats = extract_syntenic_fasta min_cov, min_pid

end

Instance Attribute Details

#genomes_list ⇒ Object (readonly)

Returns the value of attribute genomes_list.



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# File 'lib/bacterial-comparator.rb', line 18

def genomes_list
  @genomes_list
end

#stats ⇒ Object (readonly)

Returns the value of attribute stats.



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# File 'lib/bacterial-comparator.rb', line 18

def stats
  @stats
end

Instance Method Details

#build_multifasta(synteny_list) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 187

def build_multifasta synteny_list

  pep_out_dir = "#{@outdir}/align-genes-pep"

  ref_proteins = load_genome_cds(@ref_prot_file)
  synteny_list.each do |k,v|
    pep_out = File.open(pep_out_dir+"/#{k}.pep", "w")
    pep_out.write(ref_proteins[k])
    pep_out.close
  end

  @genomes_list.each_with_index do |g,i|

    genome_proteins = load_genome_cds("#{g}/Proteins.fa")
    synteny_list.each do |k,v|
      pep_out = File.open(pep_out_dir+"/#{k}.pep", "a")
      pep_out.write(genome_proteins[v[i][:query_prot]])
      pep_out.close
    end

  end

  dna_out_dir = "#{@outdir}/align-genes-dna"
  ref_genes = load_genome_cds(@ref_dna_file)
  synteny_list.each do |k,v|
    dna_out = File.open(dna_out_dir+"/#{k}.dna", "w")
    dna_out.write(ref_genes[k])
    dna_out.close
  end

  @genomes_list.each_with_index do |g,i|

    genome_genes = load_genome_cds("#{g}/Genes.fa")
    synteny_list.each do |k,v|
      dna_out = File.open(dna_out_dir+"/#{k}.dna", "a")
      dna_out.write(genome_genes[v[i][:query_prot]])
      dna_out.close
    end

  end

end

#concat_alignments(outfile, ref_id) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 415

def concat_alignments outfile, ref_id

  if File.exists?("#{outfile}") and File.size("#{outfile}") > 0
    puts "..Alignment concatenated file already exists, skipping."
    return
  end

  fout = File.open("#{outfile}", "w")

  seq = ""
  aln_dir = outfile.split(".")[0..-3].join(".")

  Dir["#{aln_dir}/*.aln"].each do |f|
    flat = Bio::FlatFile.auto(f)
    ref_seq = flat.entries[0]
    flat.close
    seq += ref_seq.seq
  end

  bioseq = Bio::Sequence.auto(seq)
  out = bioseq.output_fasta("#{ref_id}", 60)
  fout.write(out)

  for i in 1..@genomes_list.length
    seq = ""
    Dir["#{aln_dir}/*.aln"].each do |f|
      flat = Bio::FlatFile.auto(f)
      j=0
      flat.each_entry do |entry|
        if j<i
          j+=1
          next
        elsif i == j
          seq += entry.seq
          j+=1
        else
          break
        end
      end
      flat.close
    end
    bioseq = Bio::Sequence.auto(seq)
    # get the file name without path prefix and extension
    genome_name = genomes_list[i-1].split("/")[-1].split(".")[0]
    out = bioseq.output_fasta(genome_name,60)
    fout.write(out)
  end

  fout.close

end

#extract_syntenic_fasta(min_cov, min_pid) ⇒ Object

extract and dump multifasta for syntenic genes and proteins



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# File 'lib/bacterial-comparator.rb', line 231

def extract_syntenic_fasta min_cov, min_pid

  puts "# Extracting Proteins and Genes multifasta  START.."
  start_time = Time.now

  nb_of_syntenic = 0
  stats = {}
  stats[:syntenic] = []
  fout = File.open("#{@outdir}/cds-synteny.tsv", "w")
  genomes_name = []
  @genomes_list.each do |g|
    genomes_name.push(File.basename(g))
  end
  fout.write("Gene\t"+genomes_name.join("\t")+"\n")

  to_build_multifasta = []

  @synteny.each do |k,v|

    is_syntenic = 1
    v.each do |v_|
      if v_[:query_cov] == "-"
        is_syntenic = 0
        break
      elsif v_[:query_cov] > min_cov and
           v_[:ref_cov] > min_cov and
           v_[:pId] > min_pid
      # synteny -> great !
      else
        is_syntenic = 0
        break
      end
    end

    fout.write("#{k}")
    if is_syntenic == 1
      nb_of_syntenic += 1
      # build_multifasta k, v
      to_build_multifasta << [k,v]
      v.each do |x|
        fout.write("\t#{x[:query_prot]}|#{x[:pId]}|#{x[:query_cov]}|#{x[:ref_cov]}")
        stats[:syntenic] << k
      end
      fout.write("\n")
    else
      v.each do |x|
        if x[:pId] == 0.0
          fout.write("\t-")
        else
          fout.write("\t#{x[:query_prot]}|#{x[:pId]}|#{x[:query_cov]}|#{x[:ref_cov]}")
        end
      end
      fout.write("\n")
    end

  end

  fout.close

  pep_out_dir = "#{@outdir}/align-genes-pep"
  dna_out_dir = "#{@outdir}/align-genes-dna"
  Dir.mkdir(pep_out_dir) if ! Dir.exists? pep_out_dir
  Dir.mkdir(dna_out_dir) if ! Dir.exists? dna_out_dir

  synteny_list = to_build_multifasta.each_slice((to_build_multifasta.length/@proc)+1).to_a

  Parallel.map(synteny_list, in_processes: @proc) { |list|
    build_multifasta list
  }

  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  puts "# Extracting Proteins and Genes multifasta  [DONE] (in #{c_time})"

  stats[:nb_of_syntenic] = nb_of_syntenic
  #puts "   Syntenic genes : " + nb_of_syntenic.to_s + " / " + @ref_prot.length.to_s

end

#fasttree_tree_dna(bt) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 517

def fasttree_tree_dna bt
  puts "# Genes DNA tree creation (FastTree)  START.."
  start_time = Time.now
  ori_dir = Dir.pwd
  Dir.chdir(@outdir)
  Dir.mkdir("tree-genes-dna") if ! Dir.exists?("tree-genes-dna")
  current_dir = Dir.pwd
  cmd = system("export OMP_NUM_THREADS=#{@proc} && #{@root}/fasttree.linux -nosupport -fastest -nt -gtr align-genes-dna.all.fasta > tree-genes-dna.nwk")
  Dir.chdir(ori_dir)
  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  puts "# Genes DNA tree creation (FastTree)  [DONE] (in #{c_time})"
end

#fasttree_tree_pep(bt) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 532

def fasttree_tree_pep bt
  puts "# Proteins AA tree creation (FastTree)  START.."
  start_time = Time.now
  ori_dir = Dir.pwd
  Dir.chdir(@outdir)
  Dir.mkdir("tree-genes-pep") if ! Dir.exists?("tree-genes-pep")
  current_dir = Dir.pwd
  cmd = system("export OMP_NUM_THREADS=#{@proc} && #{@root}/fasttree.linux -nosupport -fastest align-genes-pep.all.fasta > tree-proteins-aa.nwk")
  Dir.chdir(ori_dir)
  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  puts "# Proteins AA tree creation (FastTree)  [DONE] (in #{c_time})"
end

#get_fasta_length(fasta) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 572

def get_fasta_length fasta
  flatfile = Bio::FlatFile.auto(fasta)
  prot_lengths = {}
  flatfile.each_entry do |entry|
    prot_id = entry.definition.split(" ")[0]
    prot_length = entry.length
    prot_lengths[prot_id] = prot_length
  end
  flatfile.close
  prot_lengths
end

#get_ref_prot ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 129

def get_ref_prot

  ref_prot = []
  if File.exist?("#{@genomes_list[0]}/*.pep")
    pep_file = Dir["#{@genomes_list[0]}/*.pep"]
    @ref_prot_file = pep_file[0]
    flatfile = Bio::FlatFile.auto("#{@ref_prot_file}")
    flatfile.each_entry do |entry|
      ref_prot << entry.definition.split(" ")[0]
    end
    flatfile.close
    dna_file = Dir["#{@genomes_list[0]}/*.dna"]
    @ref_dna_file = dna_file[0]
  else
    if @refgenome_file == ""
      abort "You need to provide a reference genome to add a non-annotated genome"
    elsif @refgenome == ""
      new_genomes_dir = @outdir+"/new-genomes/"
      Dir.mkdir(new_genomes_dir) if ! Dir.exists? new_genomes_dir
      refgenome_dir = new_genomes_dir + "/" + File.basename(@refgenome_file).gsub(".gbk","")
      Dir.mkdir(refgenome_dir) if ! Dir.exists? refgenome_dir
      @refgenome = SequenceAnnotation.new(@root,
                                          refgenome_dir,
                                          @refgenome_file,
                                          "refGbk")
      pep_file = Dir["#{refgenome_dir}/*.pep"]
      @ref_prot_file = pep_file[0]
      flatfile = Bio::FlatFile.auto("#{@ref_prot_file}")
      flatfile.each_entry do |entry|
        ref_prot << entry.definition.split(" ")[0]
      end
      flatfile.close
      dna_file = Dir["#{refgenome_dir}/*.dna"]
      @ref_dna_file = dna_file[0]
    end
  end

  ref_prot

end

#load_genome_cds(file) ⇒ Object

load all id => sequences from multifasta



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# File 'lib/bacterial-comparator.rb', line 171

def load_genome_cds file

  proteins = {}
  flatfile = Bio::FlatFile.auto(file)
  flatfile.each_entry do |entry|
    name = entry.definition.split(" ")[0]
    bioseq = Bio::Sequence.auto(entry.seq)
    out = bioseq.output_fasta("#{name}",60)
    proteins[name] = out
  end

  proteins

end

#mafft_align(f) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 310

def mafft_align f

  trying = 0
  begin
    cmd = system("#{@root}/mafft.linux --quiet #{f} > #{f}.aln")
    if File.size("#{f}.aln") == 0
      puts "File size of 0.. --#{f}--"
      puts "Command used : #{@root}/mafft.linux --quiet #{f} > #{f}.aln"
      fail
    else
      status = "OK"
      status = "FAILED" if cmd != true
      # puts "Alignment #{f} : #{status}"
    end
  rescue
    if trying < 3
      trying += 1
      retry
    end
    status = "FAILED"
    puts "Alignment #{f} : #{status}"
  end

end

#mafft_align_all_dna ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 375

def mafft_align_all_dna

  puts "# Sequence alignments - individual genes dna (MAFFT)  START.."
  start_time = Time.now

  ori_dir = Dir.pwd
  Dir.chdir("#{@outdir}/align-genes-dna/")

  is_done = 1
  if Dir["*.dna"].length == Dir["*.aln"].length
    Dir["*.aln"].each do |a|
      if File.size(a) == 0
        is_done = 0
      end
    end
  else
    is_done = 0
  end

  if is_done == 0
    Parallel.map(Dir["*.dna"], in_processes: @proc) { |f|
      mafft_align f
    }
  end

  # ugly hack to find out the reference genome FIXME
  Dir.chdir(ori_dir)
  ref_id = @ref_prot_file.split('/')[-1].gsub(".pep","")

  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  puts "# Sequence alignments - individual genes dna (MAFFT)  [DONE] (in #{c_time})"

  concat_alignments "#{@outdir}/align-genes-dna.all.fasta", ref_id

  Dir.chdir(ori_dir)

end

#mafft_align_all_pep ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 336

def mafft_align_all_pep

  puts "# Sequence alignments - individual proteins a.a. (MAFFT)  START.."
  start_time = Time.now

  ori_dir = Dir.pwd
  Dir.chdir("#{@outdir}/align-genes-pep/")

  is_done = 1
  if Dir["*.pep"].length == Dir["*.aln"].length
    Dir["*.aln"].each do |a|
      if File.size(a) == 0
        is_done = 0
      end
    end
  else
    is_done = 0
  end

  if is_done==0
    Parallel.map(Dir["*.pep"], in_processes: @proc) { |f|
      mafft_align f
    }
  end

  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  puts "# Sequence alignments - individual proteins a.a. (MAFFT)  [DONE] (in #{c_time})"

  # FIXME ugly hack to find out the reference genome
  Dir.chdir(ori_dir)
  ref_id = @ref_prot_file.split('/')[-1].gsub(".pep","")

  concat_alignments "#{@outdir}/align-genes-pep.all.fasta", ref_id

  Dir.chdir(ori_dir)

end

#raxml_tree_dna(bt) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 480

def raxml_tree_dna bt
  puts "# Genes DNA tree creation (RAXML)  START.."
  start_time = Time.now
  ori_dir = Dir.pwd
  Dir.chdir(@outdir)
  Dir.mkdir("tree-genes-dna") if ! Dir.exists?("tree-genes-dna")
  current_dir = Dir.pwd
  tree_dir = "#{current_dir}/tree-genes-dna"
  cmd = system("#{@root}/raxml.linux -T #{@proc} -f d -N #{bt} -s align-genes-dna.all.fasta  -m GTRGAMMA -p 123454321 -n DnaTree -w #{tree_dir} > /dev/null 2>&1")
  cmd = system("cat #{tree_dir}/RAxML_result.DnaTree.RUN.* >> #{tree_dir}/RAxML_result.BS")
  cmd = system("#{@root}/raxml.linux -T #{@proc} -f b -z #{tree_dir}/RAxML_result.BS -t #{tree_dir}/RAxML_bestTree.DnaTree -m GTRGAMMA -n DNA_BS_TREE -w #{tree_dir} > /dev/null 2>&1")
  cmd = system("ln -s #{tree_dir}/RAxML_bipartitionsBranchLabels.DNA_BS_TREE #{tree_dir}/../tree-genes-dna.nwk")
  Dir.chdir(ori_dir)
  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  puts "# Genes DNA tree creation (RAXML)  [DONE] (in #{c_time})"
end

#raxml_tree_pep(bt) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 498

def raxml_tree_pep bt
  puts "# Proteins AA tree creation (RAXML)  START.."
  start_time = Time.now
  ori_dir = Dir.pwd
  Dir.chdir(@outdir)
  Dir.mkdir("tree-genes-pep") if ! Dir.exists?("tree-genes-pep")
  current_dir = Dir.pwd
  tree_dir = "#{current_dir}/tree-genes-pep"
  cmd = system("#{@root}/raxml.linux -T #{@proc} -f d -N #{bt} -s align-genes-pep.all.fasta  -m PROTGAMMAAUTO -p 123454321 -n PepTree -w #{tree_dir} > /dev/null 2>&1")
  cmd = system("cat #{tree_dir}/RAxML_result.PepTree.RUN.* >> #{tree_dir}/RAxML_result.BS")
  cmd = system("#{@root}/raxml.linux -T #{@proc} -f b -z #{tree_dir}/RAxML_result.BS -t #{tree_dir}/RAxML_bestTree.PepTree -m PROTGAMMAAUTO -n PEP_BS_TREE -w #{tree_dir} > /dev/null 2>&1")
  cmd = system("ln -s #{tree_dir}/RAxML_bipartitionsBranchLabels.PEP_BS_TREE #{tree_dir}/../tree-proteins-aa.nwk")
  Dir.chdir(ori_dir)
  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  puts "# Proteins AA tree creation (RAXML)  [DONE] (in #{c_time})"
end

#read_prot_synteny ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 70

def read_prot_synteny

  puts "# Reading genome synteny files START.."
  start_time = Time.now
  synteny = {}

  # If genome is genbank (.gbk or .gb) then do syntheny first

  @genomes_list.each_with_index do |g,i|

    genome_synteny = []

    if g =~ /.gbk/i
      new_genomes_dir = @outdir+"/new-genomes/"
      Dir.mkdir(new_genomes_dir) if ! Dir.exists? new_genomes_dir
      genome_dir = new_genomes_dir + "/" + File.basename(g).gsub(".gbk","")
      Dir.mkdir(genome_dir) if ! Dir.exists? genome_dir
      genome_to_annotate = SequenceAnnotation.new(@root,
                                                  genome_dir,
                                                  g,
                                                  "refGbk")
      query_prot_file = Dir["#{genome_dir}/*.pep"][0]
      query_gene_file = Dir["#{genome_dir}/*.dna"][0]

      File.symlink(query_prot_file, File.dirname(query_prot_file)+"/Proteins.fa")
      File.symlink(query_gene_file, File.dirname(query_gene_file)+"/Genes.fa")

      run_synteny_prot @root, genome_dir, @ref_prot_file, query_prot_file

      g = genome_dir
      @genomes_list[i] = genome_dir
    end

    file = File.open("#{g}/Prot-Synteny.tsv", "r")
    l = file.gets             # skip header
    while l = file.gets
      # AAK98805.1      spr0001 453     1.0     100.0   ABAC01000005_14 453     1.0	1|0
      lA = l.chomp.split("\t")
      synteny[lA[0]] =  [] if ! synteny.has_key? lA[0]
      synteny[lA[0]] << {ref_cov: lA[3].to_f, pId: lA[4].to_f, query_prot: lA[5], query_cov: lA[7].to_f}
      genome_synteny << lA[0]
    end

    @ref_prot.each do |ref_prot|
      if ! genome_synteny.include? ref_prot
        synteny[lA[0]] << {ref_cov: "-", pId: "-", query_prot: "-", query_cov: "-"}
      end
    end
    file.close
  end
  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)

  puts "# Reading genome synteny files [DONE] (in #{c_time})"

  synteny

end

#run_comparison ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 62

def run_comparison

  run_mafft_aln

  run_phylo if @run_phylo != 0

end

#run_mafft_aln ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 467

def run_mafft_aln

  if @aln_opt == "both"
    mafft_align_all_pep
    mafft_align_all_dna
  elsif @aln_opt == "prot"
    mafft_align_all_pep
  elsif @aln_opt == "dna"
    mafft_align_all_dna
  end

end

#run_phylo ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 547

def run_phylo

  if @software == "raxml"
    if @aln_opt == "both"
      raxml_tree_dna @bootstrap
      raxml_tree_pep @bootstrap
    elsif @aln_opt == "prot"
      raxml_tree_pep @bootstrap
    elsif @aln_opt == "dna"
      raxml_tree_dna @bootstrap
    end
  elsif @software == "fasttree"
    if @aln_opt == "both"
      fasttree_tree_dna @bootstrap
      fasttree_tree_pep @bootstrap
    elsif @aln_opt == "prot"
      fasttree_tree_pep @bootstrap
    elsif @aln_opt == "dna"
      fasttree_tree_dna @bootstrap
    end
  end

end

#run_synteny_prot(root, outdir, ref_prot_file, query_prot_file) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 585

def run_synteny_prot root, outdir, ref_prot_file, query_prot_file

  puts query_prot_file
  puts ref_prot_file

  ref_synteny_prot = SequenceSynteny.new(root,
                                         outdir,
                                         query_prot_file,
                                         ref_prot_file,
                                         "Prot-Ref",
                                         0.80,
                                         0.80,
                                         "prot")

  print "# Running alignment with Reference Genome CDS (diamond).."
  start_time = Time.now
  ref_synteny_prot.run_diamond
  end_time = Time.now
  c_time = Helper.sec2str(end_time - start_time)
  print "done (#{c_time})\n"

  ref_synteny_prot.extract_hits :refgenome

  synteny_file = File.open("#{outdir}/Prot-Synteny.tsv","w")
  synteny_file.write("RefLocusTag\tRefProtID\tRefLength\tRefCoverage\tIdentity\tQueryGene\tQueryLength\tQueryCoverage\tQueryPartial\n")
  ref_annotated = {}

  ref_synteny_prot.query_sequences.each do |prot, syn_val|
    next if ! syn_val.has_key? :homology
    next if syn_val[:homology][:assert_cutoff].inject(:+) < 3
    next if ref_annotated.has_key? syn_val[:homology][:hits][0] and ref_annotated[syn_val[:homology][:hits][0]][:partial] == 0
    ref_annotated[syn_val[:homology][:hits][0]] = {
      key: prot,
      pId: syn_val[:homology][:pId],
      cov_query: syn_val[:homology][:cov_query],
      cov_subject: syn_val[:homology][:cov_subject],
      assert_cutoff: syn_val[:homology][:assert_cutoff],
      length: syn_val[:homology][:length][0],
      partial: (syn_val[:partial] ? 1 : 0)
    }
    # ref_annotated[syn_val[:homology][:hits][0]] = {
    #   key: prot,
    #   pId: syn_val[:homology][:pId],
    #   cov_query: syn_val[:homology][:cov_query],
    #   cov_subject: syn_val[:homology][:cov_subject],
    #   assert_cutoff: syn_val[:homology][:assert_cutoff],
    #   length: syn_val[:homology][:length][0],
    #   partial: (syn_val[:partial] ? 1 : 0)
    # }
  end

  # print ref_annotated
  query_lengths = get_fasta_length query_prot_file

  @refgenome.coding_seq.each do |ref_k, ref_v|
    gene = ""
    coverage_ref = ""
    coverage_query = ""
    query_length = ""
    pId = ""
    if ref_annotated[ref_v[:protId]] != nil
      gene = ref_annotated[ref_v[:protId]][:key]
      coverage_ref = ref_annotated[ref_v[:protId]][:cov_subject]
      query_length = query_lengths[ref_annotated[ref_v[:protId]][:key]]
      coverage_query = ref_annotated[ref_v[:protId]][:cov_query]
      pId = ref_annotated[ref_v[:protId]][:pId]
      partial = ref_annotated[ref_v[:protId]][:partial]
    end

    _locus_tag = ref_v[:locustag] || ""
    _seq_len = "NA"
    # _seq_len = ref_v[:bioseq].seq.length.to_s if ! ref_v[:bioseq].nil?
    _seq_len = ref_v[:length].to_s if ! ref_v[:length].nil?

    synteny_file.write(ref_v[:protId])
    synteny_file.write("\t"+_locus_tag)
    synteny_file.write("\t"+_seq_len)
    synteny_file.write("\t"+coverage_ref.to_s)
    synteny_file.write("\t"+pId.to_s)
    synteny_file.write("\t"+gene)
    synteny_file.write("\t"+query_length.to_s)
    synteny_file.write("\t"+coverage_query.to_s)
    synteny_file.write("\t"+partial.to_s)
    synteny_file.write("\n")

  end

  synteny_file.close

end