Example #1
0
    def getCopy( self ):
        """return a new copy.
        """

        new_entry = Prediction()

        new_entry.mExpand = self.mExpand 
        
        new_entry.mPredictionId = self.mPredictionId 
        new_entry.mQueryToken = self.mQueryToken 
        new_entry.mQueryFrom = self.mQueryFrom 
        new_entry.mQueryTo = self.mQueryTo 
        new_entry.mSbjctToken = self.mSbjctToken 
        new_entry.mSbjctStrand = self.mSbjctStrand 
        new_entry.mSbjctFrom = self.mSbjctFrom 
        new_entry.mSbjctTo = self.mSbjctTo 
        new_entry.mRank = self.mRank 
        new_entry.score = self.score 
        new_entry.mQueryLength = self.mQueryLength 
        new_entry.mQueryCoverage = self.mQueryCoverage 
        new_entry.mNGaps = self.mNGaps 
        new_entry.mNFrameShifts = self.mNFrameShifts 
        new_entry.mNIntrons = self.mNIntrons 
        new_entry.mNSplits = self.mNSplits 
        new_entry.mNStopCodons = self.mNStopCodons 
        new_entry.mPercentIdentity = self.mPercentIdentity 
        new_entry.mPercentSimilarity = self.mPercentSimilarity 
        new_entry.mTranslation = self.mTranslation 
        new_entry.mSbjctGenomeFrom = self.mSbjctGenomeFrom 
        new_entry.mSbjctGenomeTo = self.mSbjctGenomeTo 
        new_entry.mAlignmentString = self.mAlignmentString 
        new_entry.mQueryAli = self.mQueryAli 
        new_entry.mSbjctAli = self.mSbjctAli 

        if self.mExpand:
            new_entry.mMapPeptide2Translation = alignlib.makeAlignmentVector()
            alignlib.copyAlignment( new_entry.mMapPeptide2Translation, self.mMapPeptide2Translation)
            new_entry.mMapPeptide2Genome = Genomics.String2Alignment( new_entry.mAlignmentString) 
        else:
            new_entry.mMapPeptide2Translation = self.mMapPeptide2Translation = None
            new_entry.mMapPeptide2Genome = self.mMapPeptide2Genome = None

        return new_entry
Example #2
0
            map_row2col.clear()
            alignlib.AlignmentFormatExplicit(link.mQueryFrom, link.mQueryAli, link.mSbjctFrom, link.mSbjctAli).copy(
                map_row2col
            )

            ## test all combinations, the alignment might be a suboptimal alignment in case
            ## of repeats.
            for e1 in cds[link.mQueryToken]:
                for e2 in cds[link.mSbjctToken]:
                    tmp_map_row2col.clear()
                    if param_expand:
                        alignlib.copyAlignment(
                            tmp_map_row2col,
                            map_row2col,
                            e1.mPeptideFrom + 1,
                            e1.mPeptideTo,
                            e2.mPeptideFrom + 1,
                            e2.mPeptideTo,
                        )
                    else:
                        alignlib.copyAlignment(
                            tmp_map_row2col,
                            map_row2col,
                            e1.mPeptideFrom / 3 + 1,
                            e1.mPeptideTo / 3 + 1,
                            e2.mPeptideFrom / 3 + 1,
                            e2.mPeptideTo / 3 + 1,
                        )

                    ## in case of split codons, there is an alignment of length 1. Skip that.
                    if tmp_map_row2col.getLength() > 1:
Example #3
0
                ref_genome_from -= ref_exons_offset
                ref_genome_to   -= ref_exons_offset

                ## get percent identity for exon
                exon_percent_identity = 0
                exon_percent_similarity = 0
                
                if query_sequence and sbjct_sequence:
                    
                    tmp_ali = alignlib.makeAlignmentVector()

                    xquery_from = exon_from / 3
                    xquery_to = exon_to / 3

                    alignlib.copyAlignment( tmp_ali, entry.mMapPeptide2Translation, xquery_from, xquery_to )

                    if tmp_ali.getLength() == 0:
                        options.stdlog.write( "# WARNING: empty alignment %s\n" % str((ref_from, exon_from, ref_to, exon_to, xquery_from, xquery_to)))
                        nempty_alignments += 1
                    else:
                        if options.loglevel >= 5:
                            options.stdlog.write( "# %s\n" % str( alignlib.AlignmentFormatExplicit( tmp_ali, query_sequence, sbjct_sequence ) ) )

                        exon_percent_identity = alignlib.calculatePercentIdentity( tmp_ali,
                                                                                   query_sequence,
                                                                                   sbjct_sequence ) * 100
                        exon_percent_similarity = alignlib.calculatePercentSimilarity( tmp_ali ) * 100

                if exon_percent_identity >= min_pide:
                    is_good_exon = 1
def main( argv = None ):
    """script main.

    parses command line options in sys.argv, unless *argv* is given.
    """

    if argv == None: argv = sys.argv

    parser = E.OptionParser( version = "%prog version: $Id: gpipe/compare_predictions2exons.py 2011 2008-07-04 10:40:51Z andreas $",
                                    usage = globals()["__doc__"] )

    parser.add_option( "-g", "--genome-file", dest="genome_file", type="string",
                       help="filename with genome."  )

    parser.add_option( "-b", "--boundaries", dest="filename_boundaries", type="string",
                       help="filename with exon boundaries."  )

    parser.add_option( "-e", "--exons", dest="filename_exons", type="string",
                       help="filename with exons (output)."  )

    parser.add_option( "-p", "--peptides", dest="filename_peptides", type="string",
                       help="filename with peptide sequences."  )

    parser.add_option( "-w", "--write-notfound", dest="write_notfound", action="store_true",
                       help="print exons for predictions not found in reference."  )

    parser.add_option( "-q", "--quality-pide", dest="quality_threshold_pide", type="int",
                       help="quality threshold (pide) for exons."  )

    parser.set_defaults( 
        genome_file = "genome",
        filename_boundaries = None,
        filename_exons = None,
        filename_peptides = None,
        quality_threshold_pide = 0,
        write_notfound = False,
        ## allowed number of nucleotides for exon boundaries to
        ## be considered equivalent.
        slipping_exon_boundary = 9,
        ## stop codons to search for        
        stop_codons = ("TAG", "TAA", "TGA"), )


    (options, args) = E.Start( parser, add_pipe_options = True )

    if len(args) > 0:
        print USAGE, "no arguments required."
        sys.exit(2)

    reference_exon_boundaries = {}
    if options.filename_boundaries:
        reference_exon_boundaries = Exons.ReadExonBoundaries( open( options.filename_boundaries, "r"),
                                                              do_invert = 1,
                                                              remove_utr = 1)
        E.info( "read exon boundaries for %i queries" % len(reference_exon_boundaries) )
                
    if options.filename_exons:
        outfile_exons = open( options.filename_exons, "w")
        outfile_exons.write( "%s\n" % "\t".join( (
                    "prediction_id",
                    "exon_id",
                    "exon_from",
                    "exon_to",
                    "exon_frame",
                    "reference_id",
                    "reference_from",
                    "reference_to",
                    "reference_phase",
                    "pidentity",
                    "psimilarity",
                    "nframeshifts",
                    "ngaps",
                    "nstopcodons",
                    "is_ok",
                    "genome_exon_from",
                    "genome_exon_to") ) )

    else:
        outfile_exons = None

    if options.filename_peptides:
        peptide_sequences = Genomics.ReadPeptideSequences( open(options.filename_peptides, "r") )
        E.info("read peptide sequences for %i queries" % len(peptide_sequences) )
    else:
        peptide_sequences = {}

    entry = PredictionParser.PredictionParserEntry()
    last_filename_genome = None
    
    nfound, nmissed_exons, nmissed_length = 0, 0, 0
    nempty_alignments = 0

    fasta = IndexedFasta.IndexedFasta( options.genome_file )

    options.stdout.write( "%s\n" % "\t".join( (
                "prediction_id", 
                "number",
                "dubious_exons",
                "boundaries_sum",
                "boundaries_max",
                "identical_exons",
                "inserted_exons",
                "deleted_exons",
                "inserted_introns",
                "deleted_introns",
                "truncated_Nterminus",
                "truncated_Cterminus",
                "deleted_Nexons",
                "deleted_Cexons",
                "inserted_Nexons",
                "inserted_Cexons" ) ) )

    for line in sys.stdin:

        if line[0] == "#": continue
        
        try:
            entry.Read(line)
        except ValueError, msg:
            print "# parsing failed with msg %s in line %s" % (msg, line[:-1])
            sys.exit(1)

        exons = Genomics.Alignment2ExonBoundaries( entry.mMapPeptide2Genome,
                                                   query_from = entry.mQueryFrom,
                                                   sbjct_from = entry.mSbjctGenomeFrom,
                                                   add_stop_codon = 0 )

        if exons[-1][4] != entry.mSbjctGenomeTo:
            print "# WARNING: discrepancy in exon calculation!!!"
            for e in exons:
                print "#", str(e)
            print "#", str(entry)

        if options.loglevel >= 5:
            for e in exons:
                print "#", str(e)
        
        genomic_fragment = fasta.getSequence( entry.mSbjctToken, entry.mSbjctStrand,
                                              entry.mSbjctGenomeFrom, entry.mSbjctGenomeTo )
        
        skip = False
        if peptide_sequences.has_key( entry.mQueryToken ):
            
            query_sequence = alignlib.makeSequence(peptide_sequences[entry.mQueryToken])
            sbjct_sequence = alignlib.makeSequence(entry.mTranslation)
            
            percent_similarity, percent_identity = 0, 0
            if query_sequence.getLength() < entry.mMapPeptide2Translation.getRowTo():
                print "# WARNING: query sequence %s is too short: %i %i" % ( entry.mQueryToken,
                                                                             query_sequence.getLength(),
                                                                             entry.mMapPeptide2Translation.getRowTo())
                sys.stdout.flush()
                nmissed_length += 1
                skip = True
                
            elif sbjct_sequence.getLength() < entry.mMapPeptide2Translation.getColTo():
                print "# WARNING: sbjct sequence %s is too short: %i %i" % ( entry.mSbjctToken,
                                                                       sbjct_sequence.getLength(),
                                                                       entry.mMapPeptide2Translation.getColTo())
                sys.stdout.flush()                
                nmissed_length += 1
                skip = True
            else:
                alignlib.rescoreAlignment( entry.mMapPeptide2Translation, 
                                           query_sequence, 
                                           sbjct_sequence,
                                           alignlib.makeScorer( query_sequence, sbjct_sequence ) )
                percent_identity = alignlib.calculatePercentIdentity( entry.mMapPeptide2Translation,
                                                                      query_sequence,
                                                                      sbjct_sequence ) * 100
                percent_similarity = alignlib.calculatePercentSimilarity( entry.mMapPeptide2Translation ) * 100
                
            E.debug( "prediction %s: percent identity/similarity: before=%5.2f/%5.2f, realigned=%5.2f/%5.2f" % (
                    str(entry.mPredictionId), 
                    entry.mPercentSimilarity,
                    entry.mPercentIdentity,
                    percent_similarity,
                    percent_identity ) )
                
        else:
            query_sequence = None
            sbjct_sequence = None

        # default values
        exons_num_exons = "na"
        exons_boundaries_sum = "na"
        exons_boundaries_max = "na"
        dubious_exons = "na"

        ndeleted_exons, ninserted_exons, ndeleted_introns, ninserted_introns, nidentical_exons = 0,0,0,0,0
        truncated_Nterminal_exon, truncated_Cterminal_exon = 0,0
        ndeleted_Nexons, ndeleted_Cexons = 0, 0
        ninserted_Nexons, ninserted_Cexons = 0, 0
        
        exons_offset = exons[0][3]

        if not reference_exon_boundaries.has_key( entry.mQueryToken ):
            print "# WARNING: sequence %s has no exon boundaries" % ( entry.mQueryToken )
            sys.stdout.flush()
            nmissed_exons += 1
            skip = True
        
        if not skip:

            nfound += 1
            
            ref_exons = reference_exon_boundaries[entry.mQueryToken]

            ref_exons_offset = ref_exons[0].mGenomeFrom
            
            exons_num_exons = len(ref_exons) - len(exons)
            exons_boundaries_sum = 0
            exons_phase = 0
            exons_boundaries_max = 0
            dubious_exons = 0
            
            inserted_exons = 0
            temp_inserted_exons = 0
            
            if options.loglevel >= 3:
                for e in exons:
                    options.stdlog.write( "# %s\n" % str(e) )
                for e in ref_exons:
                    options.stdlog.write( "# %s\n" % str(e) )

            min_pide = entry.mPercentIdentity * options.quality_threshold_pide / 100

            in_sync = 0
            e,r = 0,0

            while e < len(exons) and r < len(ref_exons):

                this_e, this_r = e+1, r+1
                percent_identity = 0
                percent_similarity = 0
                is_good_exon = 0

                if options.loglevel >= 4:
                    options.stdlog.write( "# current exons: %i and %i\n" % (e, r) )
                    sys.stdout.flush()
                    
                exon_from, exon_to, exon_phase, exon_genome_from, exon_genome_to, exon_ali = exons[e][0:6]
                ref_from, ref_to, ref_phase, ref_genome_from, ref_genome_to = (ref_exons[r].mPeptideFrom,
                                                                               ref_exons[r].mPeptideTo,
                                                                               ref_exons[r].frame,
                                                                               ref_exons[r].mGenomeFrom,
                                                                               ref_exons[r].mGenomeTo)

                ref_genome_from -= ref_exons_offset
                ref_genome_to   -= ref_exons_offset

                ## get percent identity for exon
                exon_percent_identity = 0
                exon_percent_similarity = 0
                
                if query_sequence and sbjct_sequence:
                    
                    tmp_ali = alignlib.makeAlignmentVector()

                    xquery_from = exon_from / 3
                    xquery_to = exon_to / 3

                    alignlib.copyAlignment( tmp_ali, entry.mMapPeptide2Translation, xquery_from, xquery_to )

                    if tmp_ali.getLength() == 0:
                        options.stdlog.write( "# WARNING: empty alignment %s\n" % str((ref_from, exon_from, ref_to, exon_to, xquery_from, xquery_to)))
                        nempty_alignments += 1
                    else:
                        if options.loglevel >= 5:
                            options.stdlog.write( "# %s\n" % str( alignlib.AlignmentFormatExplicit( tmp_ali, query_sequence, sbjct_sequence ) ) )

                        exon_percent_identity = alignlib.calculatePercentIdentity( tmp_ali,
                                                                                   query_sequence,
                                                                                   sbjct_sequence ) * 100
                        exon_percent_similarity = alignlib.calculatePercentSimilarity( tmp_ali ) * 100

                if exon_percent_identity >= min_pide:
                    is_good_exon = 1
                else:
                    is_good_exon = 0
                    
                if e < len(exons) -1 :
                    (next_exon_from, next_exon_to, next_exon_phase,
                     next_exon_genome_from, next_exon_genome_to, next_exon_ali) = exons[e+1][0:6]
                else:
                    (next_exon_from, next_exon_to, next_exon_phase,
                     next_exon_genome_from, next_exon_genome_to, next_exon_ali) = 0, 0, 0, 0, 0, []
                    
                if r < len(ref_exons) - 1:
                    next_ref_from, next_ref_to, next_ref_phase = (ref_exons[r+1].mPeptideFrom,
                                                                  ref_exons[r+1].mPeptideTo,
                                                                  ref_exons[r+1].frame)
                else:
                    next_ref_from, next_ref_to, next_ref_phase = 0, 0, 0
                    
                if options.loglevel >= 2:
                    options.stdlog.write( "# %s\n" % "\t".join( map(str, (entry.mQueryToken,
                                                                          exon_from, exon_to, exon_phase,
                                                                          exon_genome_from, exon_genome_to,
                                                                          ref_from, ref_to, ref_phase ))))
                    sys.stdout.flush()                    

                # beware of small exons.
                # if less than options.slipping_exon_boundary: boundary is 0
                # check if end is more than options.splipping_exon_boundary apart as well.
                if exon_to - exon_from <= options.slipping_exon_boundary or \
                        ref_to - ref_from <= options.slipping_exon_boundary:
                    boundary = 0
                else:
                    boundary = options.slipping_exon_boundary
                    
                if ref_to <= exon_from + boundary and \
                   ref_to <= exon_to - options.slipping_exon_boundary:
                    ## no overlap 
                    is_good_exon = 0
                    if e == 0:
                        ndeleted_Nexons += 1
                    else:
                        ndeleted_exons += 1
                    r += 1
                    exon_from, exon_to, exon_phase, exon_genome_from, exon_genome_to = 0, 0, 0, 0, 0
                    overlap = 0
                elif exon_to <= ref_from + boundary and \
                         exon_to <= ref_to - options.slipping_exon_boundary:
                    ## no overlap
                    is_good_exon = 0
                    if r == 0:
                        ninserted_Nexons += 1
                    else:
                        ninserted_exons += 1
                    e += 1
                    ref_from, ref_to, ref_phase = 0, 0, 0
                    overlap = 0
                else:
                    ## overlap
                    overlap = 1
                    dfrom = int(math.fabs(exon_from - ref_from))
                    dto = int(math.fabs(exon_to - ref_to))

                    ## get percent identity for overlapping fragment 
                    if query_sequence and sbjct_sequence:
                        ## this the problem
                        tmp_ali = alignlib.makeAlignmentVector()
                        
                        xquery_from = max( ref_from / 3, exon_from / 3)
                        xquery_to = min(ref_to / 3, exon_to / 3)

                        alignlib.copyAlignment( tmp_ali, entry.mMapPeptide2Translation, xquery_from, xquery_to )

                        if tmp_ali.getLength() == 0:
                            options.stdlog.write( "# warning: empty alignment %s\n" % str((ref_from, exon_from, ref_to, exon_to, xquery_from, xquery_to )))
                            percent_identity = 0
                            percent_similarity = 0
                        else:
                            if options.loglevel >= 5:
                                print str( alignlib.AlignmentFormatExplicit( tmp_ali, query_sequence, sbjct_sequence ) )

                            percent_identity = alignlib.calculatePercentIdentity( tmp_ali,
                                                                                  query_sequence,
                                                                                  sbjct_sequence ) * 100
                            percent_similarity = alignlib.calculatePercentSimilarity( tmp_ali ) * 100
                            
                    if percent_identity >= min_pide:
                        is_good_exon = 1
                    else:
                        is_good_exon = 0
                        dubious_exons += 1

                    ## adjust regions for terminal exons
                    if e == 0 and r == 0 and dfrom <= (entry.mQueryFrom - 1) * 3 and dfrom > 0:
                        if is_good_exon:                        
                            truncated_Nterminal_exon = dfrom
                        dfrom = 0
                            
                    ## truncated terminal exons
                    if e == len(exons)-1 and r == len(ref_exons)-1 and dto <= (entry.mQueryLength - entry.mQueryTo) * 3 and dto > 0:
                        if is_good_exon:                        
                            truncated_Cterminal_exon = dto
                        dto = 0

                    ## do not count deviations for terminal query exons
                    if e == 0 and dfrom <= entry.mQueryFrom * 3 and dfrom > 0:
                        dfrom = 0
                            
                    if e == len(exons)-1 and dto <= (entry.mQueryLength - entry.mQueryTo) * 3 and dto > 0:
                        dto = 0

                    ## permit difference of one codon (assumed to be stop)
                    if e == len(exons)-1 and r == len(ref_exons)-1 and dto == 3:
                        dto = 0

                    ## deal with different boundary conditions:
                    if dfrom == 0 and dto == 0:
                        if is_good_exon: nidentical_exons += 1
                        e += 1
                        r += 1
                    ## next exon within this ref_exon
                    elif exon_to < ref_to and next_exon_to and next_exon_to <= ref_to + options.slipping_exon_boundary:
                        if is_good_exon: ninserted_introns += 1
                        e += 1
                        in_sync = 1
                        dto = 0
                    ## next ref_exon within this exon
                    elif ref_to < exon_to and next_ref_to and next_ref_to <= exon_to + options.slipping_exon_boundary:
                        if is_good_exon: ndeleted_introns += 1
                        r += 1
                        in_sync = 1
                        dto = 0
                    else:
                        e += 1
                        r += 1
                        if in_sync:
                            dfrom = 0

                    if is_good_exon:
                        exons_boundaries_sum += dfrom + dto
                        exons_boundaries_max = max( dfrom, exons_boundaries_max )
                        exons_boundaries_max = max( dto, exons_boundaries_max )
                    
                        
                    ###########################################################
                    ## count inserted/deleted introns and misplaced boundaries
                    ##
                    ## if exon and next_exon in ref_exon: inserted intron
                    ## if ref_exon and next_ref_exon in exon: deleted intron
                    
                if outfile_exons:

                    if genomic_fragment and exon_genome_to:
                        nintrons, nframeshifts, ngaps, nsplits, nstopcodons, disruptions = Genomics.CountGeneFeatures( exon_genome_from - entry.mSbjctGenomeFrom,
                                                                                                                       exon_ali,
                                                                                                                       genomic_fragment,
                                                                                                                       border_stop_codon = 0
                                                                                                                       )
                    else:
                        nintrons, nframeshifts, ngaps, nsplits, nstopcodons = 0, 0, 0, 0, 0

                    if exon_to == 0: this_e = 0
                    if ref_to == 0: this_r = 0
                    outfile_exons.write( string.join( map(str, (entry.mPredictionId,
                                                                this_e, exon_from, exon_to, exon_phase,
                                                                this_r, ref_from, ref_to, ref_phase,
                                                                percent_identity, percent_similarity,
                                                                nframeshifts, ngaps, nstopcodons,
                                                                is_good_exon,
                                                                exon_genome_from, exon_genome_to,
                                                                )), "\t") + "\n")
                    
            while e < len(exons):
                exon_from, exon_to, exon_phase, exon_genome_from, exon_genome_to = exons[e][0:5]
                e += 1
                ninserted_Cexons += 1

                if outfile_exons:
                    outfile_exons.write( string.join( map(str, (entry.mPredictionId, 
                                                                e, exon_from, exon_to, exon_phase,
                                                                0, 0, 0, 0,
                                                                0, 0,
                                                                0, 0, 0,
                                                                1,
                                                                exon_genome_from, exon_genome_to,
                                                                )), "\t") + "\n")
                    
            while r < len(ref_exons):
                ref_from, ref_to, ref_phase, ref_genome_from, ref_genome_to = (ref_exons[r].mPeptideFrom,
                                                                               ref_exons[r].mPeptideTo,
                                                                               ref_exons[r].frame,
                                                                               ref_exons[r].mGenomeFrom,
                                                                               ref_exons[r].mGenomeTo)
                ndeleted_Cexons += 1
                ref_genome_from -= ref_exons_offset
                ref_genome_to -= ref_exons_offset
                r += 1
                if outfile_exons:
                    outfile_exons.write( string.join( map(str, (entry.mPredictionId,
                                                                0, 0, 0, 0,
                                                                r, ref_from, ref_to, ref_phase, 
                                                                0, 0,
                                                                0, 0, 0,
                                                                0,
                                                                0, 0,
                                                                )), "\t") + "\n")
        else:
            if options.write_notfound:
                this_e = 0
                ## use prediction's identity/similarity for exons.
                ## This will still then flag stop-codons in later analysis
                percent_identity = entry.mPercentIdentity
                percent_similarity = entry.mPercentSimilarity
            
                for exon in exons:
                    this_e += 1
                    exon_from, exon_to, exon_phase, exon_genome_from, exon_genome_to, exon_ali = exon[0:6]
                    if genomic_fragment:
                        nintrons, nframeshifts, ngaps, nsplits, nstopcodons, disruptions = Genomics.CountGeneFeatures( exon_genome_from - entry.mSbjctGenomeFrom,
                                                                                                                       exon_ali,
                                                                                                                       genomic_fragment )
                    
                    outfile_exons.write( string.join( map(str, (entry.mPredictionId,
                                                                this_e, exon_from, exon_to, exon_phase,
                                                                0, 0, 0, 0,
                                                                percent_identity, percent_similarity,
                                                                nframeshifts, ngaps, nstopcodons,
                                                                1,
                                                                exon_genome_from, exon_genome_to,
                                                                )), "\t") + "\n")
            
        options.stdout.write( "\t".join(map(str,
                              (entry.mPredictionId,
                               exons_num_exons,
                               dubious_exons,
                               exons_boundaries_sum,
                               exons_boundaries_max,
                               nidentical_exons,
                               ninserted_exons, ndeleted_exons,
                               ninserted_introns, ndeleted_introns,
                               truncated_Nterminal_exon, truncated_Cterminal_exon,
                               ndeleted_Nexons, ndeleted_Cexons,
                               ninserted_Nexons, ninserted_Cexons))) + "\n" )
Example #5
0
def pslMap( options ):
    """thread psl alignments using intervals.

    """

    if options.format == "gtf":
        use_copy = False
    else:
        use_copy = True

    ninput, noutput, ndiscarded, nskipped, nskipped_small_queries = 0, 0, 0, 0, 0

    min_length = options.min_aligned

    for match, qx, tx in iterator_psl_intervals( options ):

        map_query2target = match.getMapQuery2Target()

        ninput += 1

        ## if no filter on qx or tx, use full segment
        if qx == None:
            qx = [ (match.mQueryFrom,match.mQueryTo,0) ]
        elif tx == None:
            tx = [ (match.mSbjctFrom,match.mSbjctTo,0) ]

        ## if no overlap: return
        if not qx or not tx: 
            nskipped += 1
            continue

        for query in qx:

            qstart, qend, qval = query

            # skip elements that are too small
            if qend - qstart < min_length: 
                E.debug( "query too small - skipped at %s:%i-%i" % (match.mQueryId, qstart, qend) )
                nskipped_small_queries += 1
                continue

            E.debug( "working on query %s:%i-%i" % (match.mQueryId, qstart, qend) )

            mqstart, mqend = ( map_query2target.mapRowToCol(qstart, 
                                                            alignlib.RIGHT), 
                               map_query2target.mapRowToCol(qend, 
                                                            alignlib.LEFT) )
                        
                
            if match.strand == "-":
                qstart, qend = match.mQueryLength - qend, match.mQueryLength - qstart

            for target in tx:

                tstart, tend, tval = target
                if tstart >= mqend or tend <= mqstart: continue
                if tend - tstart < min_length: continue

                new = alignlib.makeAlignmentBlocks()
                    
                if use_copy:
                    # do copy with range filter
                    if options.loglevel >= 3:

                        mtstart, mtend = map_query2target.mapColToRow(tstart), map_query2target.mapColToRow(tend) 
                        E.debug( "query: %i-%i (len=%i)-> %i-%i(len=%i); target: %i-%i (len=%i)-> %i-%i (len=%i)" % \
                                     (qstart, qend,
                                      qend - qstart,
                                      mqstart, mqend,
                                      mqend - mqstart,
                                      tstart, tend,
                                      tend - tstart,
                                      mtstart, mtend,
                                      mtend - mtstart ) )
                                     
                    alignlib.copyAlignment( 
                        new, 
                        map_query2target,
                        qstart, qend,
                        tstart, tend )
                else:
                    # do copy with alignment filter
                    map_query = qval
                    if map_query:
                        tmp = alignlib.makeAlignmentBlocks()                        
                        alignlib.copyAlignment( tmp, map_query2target, map_query, alignlib.RR )
                        if options.loglevel >= 5:
                            options.stdlog.write( "######## mapping query ###########\n" )
                            options.stdlog.write( "# %s\n" % str(alignlib.AlignmentFormatEmissions( map_query2target ) ))
                            options.stdlog.write( "# %s\n" % str(alignlib.AlignmentFormatEmissions( map_query ) ))
                            options.stdlog.write( "# %s\n" % str(alignlib.AlignmentFormatEmissions( tmp ) ))
                    else:
                        tmp = map_query2target
                        
                    map_target = tval
                    if map_target:
                        new = alignlib.makeAlignmentBlocks()
                        alignlib.copyAlignment( new, tmp, map_target, alignlib.CR )                        
                        if options.loglevel >= 5:
                            options.stdlog.write( "######## mapping target ###########\n" )
                            options.stdlog.write( "# before: %s\n" % str(alignlib.AlignmentFormatEmissions( tmp ) ))
                            options.stdlog.write( "# map   : %s\n" % str(alignlib.AlignmentFormatEmissions( map_target ) ))
                            options.stdlog.write( "# after : %s\n" % str(alignlib.AlignmentFormatEmissions( new ) ))
                    else:
                        new = tmp

                if options.loglevel >= 4:
                    E.debug("putative match with intervals: %s and %s: %i-%i" % \
                                (str(query), str(target), qstart, qend ))
                    if options.loglevel >= 5:
                        E.debug( "input : %s" % str(alignlib.AlignmentFormatEmissions( map_query2target ) ))
                        E.debug( "final : %s" % str(alignlib.AlignmentFormatEmissions( new ) ) )

                    if new.getLength() > 0:
                        n = match.copy()
                        n.fromMap( new, use_strand = True )
                        E.info( "match : %s" % (str(n)))

                if new.getNumAligned() > options.min_aligned:
                    n = match.copy()
                    n.fromMap( new, use_strand = True )
                    options.stdout.write( str(n) + "\n" )
                    noutput += 1
                else:
                    ndiscarded += 1

    E.info( "map: ninput=%i, noutput=%i, nskipped=%i, ndiscarded=%i, nsmall_queries=%i" % \
                (ninput, noutput, nskipped, ndiscarded, nskipped_small_queries) )