in progress...
[jalview.git] / forester / ruby / evoruby / lib / evo / io / parser / hmmscan_domain_extractor.rb
index b7092e0..89e0fe7 100644 (file)
@@ -1,40 +1,39 @@
 #
 # = lib/evo/io/parser/hmmscan_domain_extractor.rb - HmmscanDomainExtractor class
 #
-# Copyright::  Copyright (C) 2012 Christian M. Zmasek
-# License::    GNU Lesser General Public License (LGPL)
+# Copyright::    Copyright (C) 2017 Christian M. Zmasek
+# License::      GNU Lesser General Public License (LGPL)
 #
-# $Id:  $
-
+# Last modified: 2017/02/16
 
 require 'lib/evo/util/constants'
 require 'lib/evo/msa/msa_factory'
 require 'lib/evo/io/msa_io'
 require 'lib/evo/io/writer/fasta_writer'
 require 'lib/evo/io/parser/fasta_parser'
-
+require 'lib/evo/io/parser/hmmscan_parser'
 
 module Evoruby
-
   class HmmscanDomainExtractor
 
+    ADD_TO_CLOSE_PAIRS = 0
     def initialize
     end
 
     # raises ArgumentError, IOError, StandardError
     def parse( domain_id,
-        hmmscan_output,
-        fasta_sequence_file,
-        outfile,
-        passed_seqs_outfile,
-        failed_seqs_outfile,
-        e_value_threshold,
-        length_threshold,
-        add_position,
-        add_domain_number,
-        add_species,
-        min_linker,
-        log )
+      hmmscan_output,
+      fasta_sequence_file,
+      outfile,
+      passed_seqs_outfile,
+      failed_seqs_outfile,
+      e_value_threshold,
+      length_threshold,
+      add_position,
+      add_domain_number,
+      add_species,
+      min_linker,
+      log )
 
       Util.check_file_for_readability( hmmscan_output )
       Util.check_file_for_readability( fasta_sequence_file )
@@ -58,120 +57,213 @@ module Evoruby
       out_msa_pairs = nil
       out_msa_isolated = nil
       out_msa_singles = nil
+      out_msa_single_domains_protein_seqs = nil
+      out_msa_close_pairs_protein_seqs = nil
+      out_msa_close_pairs_only_protein_seqs = nil
+      out_msa_isolated_protein_seqs = nil
+      out_msa_isolated_only_protein_seqs = nil
+      out_msa_isolated_and_close_pair_protein_seqs = nil
       if min_linker
         out_msa_pairs = Msa.new
         out_msa_isolated = Msa.new
         out_msa_singles = Msa.new
+        out_msa_single_domains_protein_seqs = Msa.new
+        out_msa_close_pairs_protein_seqs = Msa.new
+        out_msa_close_pairs_only_protein_seqs = Msa.new
+        out_msa_isolated_protein_seqs = Msa.new
+        out_msa_isolated_only_protein_seqs = Msa.new
+        out_msa_isolated_and_close_pair_protein_seqs  = Msa.new
       end
 
       ld = Constants::LINE_DELIMITER
 
-      domain_pass_counter     = 0
-      domain_fail_counter     = 0
-      proteins_with_failing_domains = 0
+      domain_pass_counter                = 0
+      domain_fail_counter                = 0
+      proteins_with_failing_domains      = 0
+      domain_not_present_counter         = 0
+      protein_counter                    = 1
       max_domain_copy_number_per_protein = -1
       max_domain_copy_number_sequence    = ""
+      passing_target_length_sum          = 0
+      overall_target_length_sum          = 0
+      overall_target_length_min          = 10000000
+      overall_target_length_max          = 0
+      passing_target_length_min          = 10000000
+      passing_target_length_max          = 0
 
-      hmmscan_datas = Array.new
+      hmmscan_datas = []
 
-      File.open( hmmscan_output ) do | file |
-        while line = file.gets
-          if !is_ignorable?( line ) && line =~ /^\S+\s+/
+      hmmscan_parser = HmmscanParser.new( hmmscan_output )
+      results = hmmscan_parser.parse
 
-            #         tn      acc     tlen    query   acc     qlen    Evalue  score   bias    #       of      c-E     i-E     score   bias    hf      ht      af      at      ef      et      acc     desc
-            #         1       2       3       4       5       6       7       8       9       10      11      12      13      14      15      16      17      18      19      20      21      22      23
-            line =~ /^(\S+)\s+(\S+)\s+(\d+)\s+(\S+)\s+(\S+)\s+(\d+)\s+(\S+)\s+(\S+)\s+(\S+)\s+(\d+)\s+(\d+)\s+(\S+)\s+(\S+)\s+(\S+)\s+(\S+)\s+(\d+)\s+(\d+)\s+(\d+)\s+(\d+)\s+(\d+)\s+(\d+)\s+(\S+)\s+(.*)/
+      prev_query = nil
+      saw_target = false
 
-            if domain_id != $1
-              next
-            end
+      results.each do | r |
 
-            sequence = $4
-            number   = $10.to_i
-            out_of   = $11.to_i
-            env_from = $20.to_i
-            env_to   = $21.to_i
-            i_e_value  = $13.to_f
-
-            if ( ( ( e_value_threshold < 0.0 ) || ( i_e_value <= e_value_threshold ) ) &&
-                 ( ( length_threshold <= 0 )   || ( env_to - env_from + 1 ) >= length_threshold.to_f ) )
-              hmmscan_datas << HmmsearchData.new( sequence, number, out_of, env_from, env_to, i_e_value )
-              if ( number > max_domain_copy_number_per_protein )
-                max_domain_copy_number_sequence    = sequence
-                max_domain_copy_number_per_protein = number
-              end
-            else # failed
-              print( domain_fail_counter.to_s + ": " + sequence.to_s + " did not meet threshold(s)" )
-              log << domain_fail_counter.to_s + ": " + sequence.to_s + " did not meet threshold(s)"
-              if ( ( e_value_threshold.to_f >= 0.0 ) && ( i_e_value > e_value_threshold ) )
-                print( " iE=" + i_e_value.to_s )
-                log << " iE=" + i_e_value.to_s
-              end
-              if ( ( length_threshold.to_f > 0 ) && ( env_to - env_from + 1 ) < length_threshold.to_f )
-                le = env_to - env_from + 1
-                print( " l=" + le.to_s )
-                log << " l=" + le.to_s
-              end
-              print( ld )
-              log << ld
-              domain_fail_counter  += 1
-            end
+        if ( prev_query != nil ) && ( r.query != prev_query )
+          protein_counter += 1
+          if !saw_target
+            log << domain_not_present_counter.to_s + ": " + prev_query.to_s + " lacks target domain" + ld
+            domain_not_present_counter += 1
+          end
+          saw_target = false
+        end
+
+        prev_query = r.query
+
+        if domain_id != r.model
+          next
+        end
+
+        saw_target = true
+
+        sequence  = r.query
+        number    = r.number
+        out_of    = r.out_of
+        env_from  = r.env_from
+        env_to    = r.env_to
+        i_e_value = r.i_e_value
+        prev_query = r.query
 
-            if number > out_of
-              error_msg = "number > out_of ! (this should not have happened)"
+        length = env_to - env_from + 1
+
+        overall_target_length_sum += length
+        if length > overall_target_length_max
+          overall_target_length_max = length
+        end
+        if length < overall_target_length_min
+          overall_target_length_min = length
+        end
+
+        if ( ( ( e_value_threshold < 0.0 ) || ( i_e_value <= e_value_threshold ) ) &&
+        ( ( length_threshold <= 0 ) || ( length >= length_threshold.to_f ) ) )
+          hmmscan_datas << HmmsearchData.new( sequence, number, out_of, env_from, env_to, i_e_value )
+          passing_target_length_sum += length
+          if length > passing_target_length_max
+            passing_target_length_max = length
+          end
+          if length < passing_target_length_min
+            passing_target_length_min = length
+          end
+          if ( number > max_domain_copy_number_per_protein )
+            max_domain_copy_number_sequence    = sequence
+            max_domain_copy_number_per_protein = number
+          end
+        else # no pass
+          log << domain_fail_counter.to_s + ": " + sequence.to_s + " fails threshold(s)"
+          if ( ( e_value_threshold.to_f >= 0.0 ) && ( i_e_value > e_value_threshold ) )
+            log << " iE=" + i_e_value.to_s
+          end
+          if ( ( length_threshold.to_f > 0 ) && ( env_to - env_from + 1 ) < length_threshold.to_f )
+            le = env_to - env_from + 1
+            log << " l=" + le.to_s
+          end
+          log << ld
+          domain_fail_counter += 1
+        end
+
+        if number > out_of
+          error_msg = "number > out_of (this should not have happened)"
+          raise StandardError, error_msg
+        end
+
+        if number == out_of
+          if !hmmscan_datas.empty?
+            process_hmmscan_datas( hmmscan_datas,
+            in_msa,
+            add_position,
+            add_domain_number,
+            add_species,
+            out_msa,
+            out_msa_singles,
+            out_msa_pairs,
+            out_msa_isolated,
+            min_linker,
+            out_msa_single_domains_protein_seqs,
+            out_msa_close_pairs_protein_seqs,
+            out_msa_close_pairs_only_protein_seqs,
+            out_msa_isolated_protein_seqs,
+            out_msa_isolated_only_protein_seqs,
+            out_msa_isolated_and_close_pair_protein_seqs )
+            domain_pass_counter += hmmscan_datas.length
+            if passed_seqs.find_by_name_start( sequence, true ).length < 1
+              add_sequence( sequence, in_msa, passed_seqs )
+            else
+              error_msg = "this should not have happened"
               raise StandardError, error_msg
             end
-
-            if number == out_of
-              if !hmmscan_datas.empty?
-                process_hmmscan_datas( hmmscan_datas,
-                  in_msa,
-                  add_position,
-                  add_domain_number,
-                  add_species,
-                  out_msa,
-                  out_msa_singles,
-                  out_msa_pairs,
-                  out_msa_isolated,
-                  min_linker )
-                domain_pass_counter += hmmscan_datas.length
-                if passed_seqs.find_by_name_start( sequence, true ).length < 1
-                  add_sequence( sequence, in_msa, passed_seqs )
-                else
-                  error_msg = "this should not have happened"
-                  raise StandardError, error_msg
-                end
-              else
-                if failed_seqs.find_by_name_start( sequence, true ).length < 1
-                  add_sequence( sequence, in_msa, failed_seqs )
-                  proteins_with_failing_domains += 1
-                else
-                  error_msg = "this should not have happened"
-                  raise StandardError, error_msg
-                end
-              end
-              hmmscan_datas.clear
+          else # no pass
+            if failed_seqs.find_by_name_start( sequence, true ).length < 1
+              add_sequence( sequence, in_msa, failed_seqs )
+              proteins_with_failing_domains += 1
+            else
+              error_msg = "this should not have happened"
+              raise StandardError, error_msg
             end
-
           end
-        end #  while line = file.gets
-      end # File.open( hmmsearch_output ) do | file |
+          hmmscan_datas.clear
+        end
 
+      end # results.each do | r |
 
+      if (prev_query != nil) && (!saw_target)
+        log << domain_not_present_counter.to_s + ": " + prev_query.to_s + " lacks target domain" + ld
+        domain_not_present_counter += 1
+      end
 
       if domain_pass_counter < 1
         error_msg = "no domain sequences were extracted"
         raise IOError, error_msg
       end
 
+      if ( domain_not_present_counter + passed_seqs.get_number_of_seqs + proteins_with_failing_domains ) != protein_counter
+        error_msg = "not present + passing + not passing != proteins in sequence (fasta) file (this should not have happened)"
+        raise StandardError, error_msg
+      end
+
+      puts
+      log << ld
+
+      log << ld
+      avg_pass = ( passing_target_length_sum / domain_pass_counter )
+      puts( "Passing target domain lengths: average: " + avg_pass.to_s  )
+      log << "Passing target domain lengths: average: " + avg_pass.to_s
+      log << ld
+      puts( "Passing target domain lengths: min-max: " + passing_target_length_min.to_s + "-"  + passing_target_length_max.to_s)
+      log << "Passing target domain lengths: min-max: " + passing_target_length_min.to_s + "-"  + passing_target_length_max.to_s
+      log << ld
+      puts( "Passing target domain lengths:     sum: " + domain_pass_counter.to_s  )
+      log << "Passing target domain lengths:     sum: " + domain_pass_counter.to_s
+      log << ld
+      log << ld
+      puts
+      sum = domain_pass_counter + domain_fail_counter
+      avg_all = overall_target_length_sum / sum
+      puts( "All target domain lengths:     average: " + avg_all.to_s  )
+      log << "All target domain lengths:     average: " +avg_all.to_s
+      log << ld
+      puts( "All target domain lengths:     min-max: " + overall_target_length_min.to_s + "-"  + overall_target_length_max.to_s)
+      log << "All target domain lengths:     min-max: " + overall_target_length_min.to_s + "-"  + overall_target_length_max.to_s
+      log << ld
+      puts( "All target domain lengths:         sum: " + sum.to_s  )
+      log << "All target domain lengths:         sum: " + sum.to_s
+
+      puts
+      puts( "Proteins with passing target domain(s): " + passed_seqs.get_number_of_seqs.to_s )
+      puts( "Proteins with no passing target domain: " + proteins_with_failing_domains.to_s )
+      puts( "Proteins with no target domain        : " + domain_not_present_counter.to_s )
+
       log << ld
-      puts( "Max domain copy number per protein : " + max_domain_copy_number_per_protein.to_s )
-      log << "Max domain copy number per protein : " + max_domain_copy_number_per_protein.to_s
+      log << ld
+      puts
+      puts( "Max target domain copy number per protein (includes non-passing): " + max_domain_copy_number_per_protein.to_s )
+      log << "Max target domain copy number per protein (includes non-passing): " + max_domain_copy_number_per_protein.to_s
       log << ld
 
       if ( max_domain_copy_number_per_protein > 1 )
-        puts( "First protein with this copy number: " + max_domain_copy_number_sequence )
-        log << "First protein with this copy number: " + max_domain_copy_number_sequence
+        puts( "First target protein with this copy number: " + max_domain_copy_number_sequence )
+        log << "First target protein with this copy number: " + max_domain_copy_number_sequence
         log << ld
       end
 
@@ -180,35 +272,78 @@ module Evoruby
       write_msa( failed_seqs, failed_seqs_outfile )
 
       if out_msa_pairs
-        write_msa( out_msa_pairs, outfile +"_" + min_linker.to_s + ".fasta")
+        write_msa( out_msa_pairs, outfile + "_" + min_linker.to_s + ".fasta")
       end
 
       if out_msa_singles
-        write_msa( out_msa_singles, outfile +"_singles" + ".fasta")
+        write_msa( out_msa_singles, outfile + "_singles.fasta")
       end
 
       if out_msa_isolated
-        write_msa( out_msa_isolated, outfile + "_" + min_linker.to_s + "_isolated" + ".fasta");
+        write_msa( out_msa_isolated, outfile + "_" + min_linker.to_s + "_isolated.fasta");
+      end
+
+      if out_msa_single_domains_protein_seqs
+        write_msa( out_msa_single_domains_protein_seqs, outfile + "_proteins_with_singles.fasta" )
+      end
+
+      if out_msa_close_pairs_protein_seqs
+        write_msa( out_msa_close_pairs_protein_seqs, outfile + "_" + min_linker.to_s + "_proteins_close_pairs.fasta" )
+      end
+
+      if out_msa_close_pairs_only_protein_seqs
+        write_msa( out_msa_close_pairs_only_protein_seqs, outfile + "_" + min_linker.to_s + "_proteins_with_close_pairs_only.fasta" )
+      end
+
+      if  out_msa_isolated_protein_seqs
+        write_msa(  out_msa_isolated_protein_seqs, outfile + "_" + min_linker.to_s + "_proteins_with_isolated_domains.fasta" )
+      end
+
+      if out_msa_isolated_only_protein_seqs
+        write_msa( out_msa_isolated_only_protein_seqs, outfile + "_" + min_linker.to_s + "_proteins_with_isolated_domains_only.fasta" )
       end
 
+      if out_msa_isolated_and_close_pair_protein_seqs
+        write_msa( out_msa_isolated_and_close_pair_protein_seqs, outfile + "_" + min_linker.to_s + "_proteins_with_isolated_and_close_pairs.fasta" )
+      end
+
+      if min_linker
+        if ( out_msa_single_domains_protein_seqs.get_number_of_seqs +
+        out_msa_close_pairs_only_protein_seqs.get_number_of_seqs +
+        out_msa_isolated_only_protein_seqs.get_number_of_seqs +
+        out_msa_isolated_and_close_pair_protein_seqs.get_number_of_seqs ) != passed_seqs.get_number_of_seqs
+          error_msg = "this should not have happened"
+          raise StandardError, error_msg
+        end
+      end
 
       log << ld
-      log << "passing domains                 : " + domain_pass_counter.to_s + ld
-      if ( min_linker )
-        log << "single domains                  : " + out_msa_singles.get_number_of_seqs.to_s + ld
-        log << "domains in close pairs          : " + out_msa_pairs.get_number_of_seqs.to_s + ld
-        log << "isolated domains                : " + out_msa_isolated.get_number_of_seqs.to_s + ld
+      log << "passing target domains                       : " + domain_pass_counter.to_s + ld
+      log << "failing target domains                       : " + domain_fail_counter.to_s + ld
+      log << "proteins in sequence (fasta) file            : " + in_msa.get_number_of_seqs.to_s + ld
+      log << "proteins in hmmscan outputfile               : " + protein_counter.to_s + ld
+      log << "proteins with passing target domain(s)       : " + passed_seqs.get_number_of_seqs.to_s + ld
+      log << "proteins with no passing target domain       : " + proteins_with_failing_domains.to_s + ld
+      log << "proteins with no target domain               : " + domain_not_present_counter.to_s + ld
+      if min_linker
+        log << "min linker length                            : " + min_linker.to_s + ld
+        log << "single domains                               : " + out_msa_singles.get_number_of_seqs.to_s + ld
+        log << "domains in close pairs                       : " + (2 * out_msa_pairs.get_number_of_seqs).to_s + ld
+        log << "isolated domains                             : " + out_msa_isolated.get_number_of_seqs.to_s + ld
+        log << "proteins with single domains                 : " + out_msa_single_domains_protein_seqs.get_number_of_seqs.to_s + ld
+        log << "proteins with close pair domains             : " + out_msa_close_pairs_protein_seqs.get_number_of_seqs.to_s + ld
+        log << "proteins with close pair domains only        : " + out_msa_close_pairs_only_protein_seqs.get_number_of_seqs.to_s + ld
+        log << "proteins with isolated domains               : " + out_msa_isolated_protein_seqs.get_number_of_seqs.to_s + ld
+        log << "proteins with isolated domains only          : " + out_msa_isolated_only_protein_seqs.get_number_of_seqs.to_s + ld
+        log << "proteins with close pair and isolated domains: " + out_msa_isolated_and_close_pair_protein_seqs.get_number_of_seqs.to_s + ld
       end
-      log << "failing domains                 : " + domain_fail_counter.to_s + ld
-      log << "proteins with passing domains   : " + passed_seqs.get_number_of_seqs.to_s + ld
-      log << "proteins with no passing domains: " + proteins_with_failing_domains.to_s + ld
+
       log << ld
 
       return domain_pass_counter
 
     end # parse
 
-
     private
 
     def write_msa( msa, filename )
@@ -224,7 +359,6 @@ module Evoruby
       end
     end
 
-
     def add_sequence( sequence_name, in_msa, add_to_msa )
       seqs = in_msa.find_by_name_start( sequence_name, true )
       if ( seqs.length < 1 )
@@ -240,17 +374,28 @@ module Evoruby
     end
 
     def process_hmmscan_datas( hmmscan_datas,
-        in_msa,
-        add_position,
-        add_domain_number,
-        add_species,
-        out_msa,
-        out_msa_singles,
-        out_msa_pairs,
-        out_msa_isolated,
-        min_linker )
+      in_msa,
+      add_position,
+      add_domain_number,
+      add_species,
+      out_msa,
+      out_msa_singles,
+      out_msa_pairs,
+      out_msa_isolated,
+      min_linker,
+      out_msa_single_domains_protein_seqs,
+      out_msa_close_pairs_protein_seqs,
+      out_msa_close_pairs_only_protein_seqs,
+      out_msa_isolated_protein_seqs,
+      out_msa_isolated_only_protein_seqs,
+      out_msa_isolated_and_close_pair_protein_seqs )
 
       actual_out_of = hmmscan_datas.size
+      saw_close_pair = false
+      saw_isolated = false
+
+      seq_name = ""
+      prev_seq_name = nil
 
       hmmscan_datas.each_with_index do |hmmscan_data, index|
         if hmmscan_data.number < ( index + 1 )
@@ -258,82 +403,153 @@ module Evoruby
           raise StandardError, error_msg
         end
 
-        extract_domain( hmmscan_data.seq_name,
-          index + 1,
-          actual_out_of,
-          hmmscan_data.env_from,
-          hmmscan_data.env_to,
-          in_msa,
-          out_msa,
-          add_position,
-          add_domain_number,
-          add_species )
+        seq_name =  hmmscan_data.seq_name
+
+        extract_domain( seq_name,
+        index + 1,
+        actual_out_of,
+        hmmscan_data.env_from,
+        hmmscan_data.env_to,
+        in_msa,
+        out_msa,
+        add_position,
+        add_domain_number,
+        add_species )
 
         if min_linker
           if actual_out_of == 1
-            extract_domain( hmmscan_data.seq_name,
-              1,
-              1,
+            extract_domain( seq_name,
+            1,
+            1,
+            hmmscan_data.env_from,
+            hmmscan_data.env_to,
+            in_msa,
+            out_msa_singles,
+            add_position,
+            add_domain_number,
+            add_species )
+            if out_msa_single_domains_protein_seqs.find_by_name_start( seq_name, true ).length < 1
+              add_sequence( seq_name, in_msa, out_msa_single_domains_protein_seqs )
+            else
+              error_msg = "this should not have happened"
+              raise StandardError, error_msg
+            end
+
+          else
+            first = index == 0
+            last = index == hmmscan_datas.length - 1
+
+            if ( ( first && ( ( hmmscan_datas[ index + 1 ].env_from - hmmscan_data.env_to ) > min_linker) )  ||
+            ( last && ( ( hmmscan_data.env_from - hmmscan_datas[ index - 1 ].env_to ) > min_linker ) ) ||
+            ( !first && !last &&  ( ( hmmscan_datas[ index + 1 ].env_from - hmmscan_data.env_to ) > min_linker ) &&
+            ( ( hmmscan_data.env_from - hmmscan_datas[ index - 1 ].env_to ) > min_linker ) ) )
+
+              extract_domain( seq_name,
+              index + 1,
+              actual_out_of,
               hmmscan_data.env_from,
               hmmscan_data.env_to,
               in_msa,
-              out_msa_singles,
+              out_msa_isolated,
               add_position,
               add_domain_number,
               add_species )
-          else
-            first = index == 0
-            last = index == hmmscan_datas.length - 1
-
-            if ( ( first && ( ( hmmscan_datas[ index + 1 ].env_from - hmmscan_data.env_to ) > min_linker) )  ||
-                 ( last && ( ( hmmscan_data.env_from - hmmscan_datas[ index - 1 ].env_to ) > min_linker ) ) ||
-                 ( !first && !last &&  ( ( hmmscan_datas[ index + 1 ].env_from - hmmscan_data.env_to ) > min_linker ) &&
-                   ( ( hmmscan_data.env_from - hmmscan_datas[ index - 1 ].env_to ) > min_linker ) ) )
-
-              extract_domain(  hmmscan_data.seq_name,
-                index + 1,
-                actual_out_of,
-                hmmscan_data.env_from,
-                hmmscan_data.env_to,
-                in_msa,
-                out_msa_isolated,
-                add_position,
-                add_domain_number,
-                add_species )
+              saw_isolated = true
 
             elsif !first
-              extract_domain(  hmmscan_data.seq_name,
-                index.to_s  + "+" + ( index + 1 ).to_s,
-                actual_out_of,
-                hmmscan_datas[ index - 1 ].env_from,
-                hmmscan_data.env_to,
-                in_msa,
-                out_msa_pairs,
-                add_position,
-                add_domain_number,
-                add_species )
+
+              from = hmmscan_datas[ index - 1 ].env_from
+              to = hmmscan_data.env_to
+
+              if ADD_TO_CLOSE_PAIRS > 0
+                from = from - ADD_TO_CLOSE_PAIRS
+                if from < 1
+                  from = 1
+                end
+                to = to + ADD_TO_CLOSE_PAIRS
+                temp_seqs = in_msa.find_by_name_start( seq_name, true )
+                temp_seq = in_msa.get_sequence( temp_seqs[ 0 ] )
+                if to >  temp_seq.get_length
+                  to =  temp_seq.get_length
+                end
+              end
+
+              extract_domain( seq_name,
+              index.to_s  + "+" + ( index + 1 ).to_s,
+              actual_out_of,
+              from,
+              to,
+              in_msa,
+              out_msa_pairs,
+              add_position,
+              add_domain_number,
+              add_species )
+              saw_close_pair = true
             end
           end
         end
+        if prev_seq_name && prev_seq_name != seq_name
+          error_msg = "this should not have happened"
+          raise StandardError, error_msg
+        end
+        prev_seq_name = seq_name
       end # each
+      if saw_isolated
+        if out_msa_isolated_protein_seqs.find_by_name_start( seq_name, true ).length < 1
+          add_sequence( seq_name, in_msa, out_msa_isolated_protein_seqs )
+        else
+          error_msg = "this should not have happened"
+          raise StandardError, error_msg
+        end
+      end
+      if saw_close_pair
+        if out_msa_close_pairs_protein_seqs.find_by_name_start( seq_name, true ).length < 1
+          add_sequence( seq_name, in_msa, out_msa_close_pairs_protein_seqs )
+        else
+          error_msg = "this should not have happened"
+          raise StandardError, error_msg
+        end
+      end
+      if saw_close_pair && saw_isolated
+        if out_msa_isolated_and_close_pair_protein_seqs.find_by_name_start( seq_name, true ).length < 1
+          add_sequence( seq_name, in_msa, out_msa_isolated_and_close_pair_protein_seqs )
+        else
+          error_msg = "this should not have happened"
+          raise StandardError, error_msg
+        end
+      elsif saw_close_pair
+        if out_msa_close_pairs_only_protein_seqs.find_by_name_start( seq_name, true ).length < 1
+          add_sequence( seq_name, in_msa, out_msa_close_pairs_only_protein_seqs )
+        else
+          error_msg = "this should not have happened"
+          raise StandardError, error_msg
+        end
+      elsif saw_isolated
+        if out_msa_isolated_only_protein_seqs.find_by_name_start( seq_name, true ).length < 1
+          add_sequence( seq_name, in_msa, out_msa_isolated_only_protein_seqs )
+        else
+          error_msg = "this should not have happened"
+          raise StandardError, error_msg
+        end
+      end
     end # def process_hmmscan_data
 
     def extract_domain( sequence,
-        number,
-        out_of,
-        seq_from,
-        seq_to,
-        in_msa,
-        out_msa,
-        add_position,
-        add_domain_number,
-        add_species )
-      if  number.is_a?( Fixnum ) && ( number < 1 || out_of < 1 || number > out_of )
-        error_msg = "impossible: number=" + number.to_s + ", out of=" + out_of.to_s
+      number,
+      out_of,
+      seq_from,
+      seq_to,
+      in_msa,
+      out_msa,
+      add_position,
+      add_domain_number,
+      add_species )
+      if number.is_a?( Fixnum ) && ( number < 1 || out_of < 1 || number > out_of )
+        error_msg = "number=" + number.to_s + ", out of=" + out_of.to_s
         raise StandardError, error_msg
       end
-      if  seq_from < 1 || seq_to < 1 || seq_from >= seq_to
-        error_msg = "impossible: seq-f=" + seq_from.to_s + ", seq-t=" + seq_to.to_s
+      if seq_from < 1 || seq_to < 1 || seq_from >= seq_to
+        error_msg = "impossible: seq-from=" + seq_from.to_s + ", seq-to=" + seq_to.to_s
         raise StandardError, error_msg
       end
       seqs = in_msa.find_by_name_start( sequence, true )
@@ -345,7 +561,7 @@ module Evoruby
         error_msg = "sequence \"" + sequence + "\" not unique in sequence file"
         raise IOError, error_msg
       end
-      # hmmsearch is 1 based, wheres sequences are 0 bases in this package.
+      # hmmscan is 1 based, whereas sequences are 0 bases in this package.
       seq = in_msa.get_sequence( seqs[ 0 ] ).get_subsequence( seq_from - 1, seq_to - 1 )
 
       orig_name = seq.get_name