# -*-Perl-*- Test Harness script for Bioperl
# $Id$
use strict;
BEGIN {
use lib '.';
use Bio::Root::Test;
test_begin(-tests => 282 );
use_ok('Bio::SeqIO::genbank');
}
my $verbose = test_debug();
my $ast = Bio::SeqIO->new(-format => 'genbank' ,
-verbose => $verbose,
-file => test_input_file('roa1.genbank'));
isa_ok($ast, 'Bio::SeqIO');
$ast->verbose($verbose);
my $as = $ast->next_seq();
is $as->molecule, 'mRNA',$as->accession_number;
is $as->alphabet, 'dna';
is $as->division, 'EST';
is join(',',$as->get_dates), '27-OCT-1998';
is($as->primary_id, 3598416);
my @class = $as->species->classification;
is $class[$#class],'Eukaryota';
$ast = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('NT_021877.gbk'));
$ast->verbose($verbose);
$as = $ast->next_seq();
is $as->molecule, 'DNA',$as->accession_number;
is $as->alphabet, 'dna';
is $as->division, 'CON';
is join(',',$as->get_dates), '17-OCT-2003';
is($as->primary_id, 37539616);
is($as->accession_number, 'NT_021877');
my ($cds) = grep { $_->primary_tag eq 'CDS' } $as->get_SeqFeatures();
is(($cds->get_tag_values('transl_except'))[1],
'(pos:complement(4224..4226),aa:OTHER)');
# test for a DBSOURCE line
$ast = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('BAB68554.gb'));
$ast->verbose($verbose);
$as = $ast->next_seq();
is $as->molecule, 'PRT',$as->accession_number;
is $as->alphabet, 'protein';
is $as->division, 'VRT';
is join(',',$as->get_dates), '11-APR-2002';
# Though older GenBank releases indicate SOURCE contains only the common name,
# this is no longer true. In general, this line will contain an abbreviated
# form of the full organism name (but may contain the full length name),
# as well as the optional common name and organelle. There is no get/set
# for the abbreviated name but it is accessible via name()
ok defined($as->species->name('abbreviated')->[0]);
is $as->species->name('abbreviated')->[0], 'Aldabra giant tortoise';
is($as->primary_id, 15824047);
my $ac = $as->annotation;
ok defined $ac;
my @dblinks = $ac->get_Annotations('dblink');
is(scalar @dblinks,1);
is($dblinks[0]->database, 'GenBank');
is($dblinks[0]->primary_id, 'AB072353');
is($dblinks[0]->version, '1');
is($dblinks[0]->display_text, 'GenBank:AB072353.1','operator overloading in AnnotationI is deprecated');
# test for multi-line SOURCE
$ast = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('NC_006346.gb'));
$as = $ast->next_seq;
is $as->species->binomial('FULL'), 'Bolitoglossa n. sp. RLM-2004',$as->accession_number;;
@class = $as->species->classification;
is($class[$#class],'Eukaryota');
is($as->species->common_name,'mushroomtongue salamander');
$ast = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('U71225.gb'));
$as = $ast->next_seq;
@class = $as->species->classification;
is($class[$#class],'Eukaryota',$as->accession_number);
is $as->species->common_name,'black-bellied salamander';
# test for unusual common name
$ast = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('AB077698.gb'));
$as = $ast->next_seq;
# again, this is not a common name but is in name('abbreviated')
ok defined($as->species->name('abbreviated')->[0]),$as->accession_number;
is $as->species->name('abbreviated')->[0],'Homo sapiens cDNA to mRNA';
# test for common name with parentheses
$ast = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('DQ018368.gb'));
$as = $ast->next_seq;
is $as->species->scientific_name,'(Populus tomentosa x P. bolleana) x P. tomentosa var. truncata',
$as->accession_number;;
# test secondary accessions
my $seqio = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('D10483.gbk'));
my $seq = $seqio->next_seq;
my @kw = $seq->get_keywords;
is(scalar @kw, 118, $seq->accession_number);
is($kw[-1], 'yabO');
my @sec_acc = $seq->get_secondary_accessions();
is(scalar @sec_acc,14);
is($sec_acc[-1], 'X56742');
# bug #1487
my $str = Bio::SeqIO->new(-verbose => $verbose,
-file => test_input_file('D12555.gbk'));
eval {
$seq = $str->next_seq;
};
ok(! $@, 'bug 1487');
# bug 1647 rpt_unit sub-feature with multiple parens
$str = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('mini-AE001405.gb'));
ok($seq = $str->next_seq);
my @rpts = grep { $_->primary_tag eq 'repeat_region' }
$seq->get_SeqFeatures;
is $#rpts, 2, 'bug 1647';
my @rpt_units = grep {$_->has_tag('rpt_unit')} @rpts;
is $#rpt_units, 0;
is(($rpt_units[0]->get_tag_values('rpt_unit'))[0],'(TG)10;A;(TG)7');
# test bug #1673 , RDB-II genbank files
$str = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('Mcjanrna_rdbII.gbk')
);
ok($seq = $str->next_seq, 'bug 1673');
my @refs = $seq->annotation->get_Annotations('reference');
is(@refs, 1);
is($seq->display_id,'Mc.janrrnA');
is($seq->molecule ,'RNA');
is $as->division, 'PLN';
is join(',',$as->get_dates), '23-MAY-2005';
$str = Bio::SeqIO->new(-format => 'genbank',
-file => test_input_file('AF165282.gb'),
-verbose => $verbose);
$seq = $str->next_seq;
my @features = $seq->all_SeqFeatures();
is(@features, 5, $seq->accession_number);
is($features[0]->start, 1);
is($features[0]->end, 226);
my $location = $features[1]->location;
ok($location->isa('Bio::Location::SplitLocationI'));
my @sublocs = $location->sub_Location();
is(@sublocs, 29);
# version and primary ID - believe it or not, this wasn't working
is ($seq->version, 1);
is ($seq->seq_version, 1);
is ($seq->primary_id, "5734104");
# streaming and Bio::RichSeq creation
my $stream = Bio::SeqIO->new(-file => test_input_file('test.genbank'),
-verbose => $verbose,
-format => 'genbank');
$stream->verbose($verbose);
my $seqnum = 0;
my $species;
my @cl;
my $lasts;
my @ids = qw(DDU63596 DDU63595 HUMBDNF);
my @tids = (44689, 44689, 9606);
my @tnames = ("Dictyostelium discoideum","Dictyostelium discoideum",
"Homo sapiens");
while($seq = $stream->next_seq()) {
if($seqnum < 3) {
is $seq->display_id(), $ids[$seqnum];
$species = $seq->species();
@cl = $species->classification();
is( $species->binomial(), $tnames[$seqnum],
'species parsing incorrect for genbank');
is( $cl[3] ne $species->genus(), 1,
'genus duplicated in genbank parsing');
is( $species->ncbi_taxid, $tids[$seqnum] );
}
$seqnum++;
$lasts = $seq;
}
is($seqnum, 5,'streaming');
is $lasts->display_id(), "HUMBETGLOA";
my ($ref) = $lasts->annotation->get_Annotations('reference');
is($ref->medline, 94173918);
$stream->close();
$stream = Bio::SeqIO->new(-file => test_input_file('test.genbank.noseq'),
-verbose => $verbose,
-format => 'genbank' );
$seqnum = 0;
while($seq = $stream->next_seq()) {
if($seqnum < 3) {
is $seq->display_id(), $ids[$seqnum];
} elsif( $seq->display_id eq 'M37762') {
is( ($seq->get_keywords())[0], 'neurotrophic factor');
}
$seqnum++;
}
is $seqnum, 5, "Total number of sequences in test file";
# fuzzy
$seq = Bio::SeqIO->new( -format => 'genbank',
-verbose => $verbose,
-file =>test_input_file('testfuzzy.genbank'));
$seq->verbose($verbose);
ok(defined($as = $seq->next_seq()));
@features = $as->all_SeqFeatures();
is(@features,21,'Fuzzy in');
my $lastfeature = pop @features;
# this is a split location; the root doesn't have strand
is($lastfeature->strand, undef);
$location = $lastfeature->location;
#$location->verbose(-1); # silence the warning of undef seq_id()
# see above; splitlocs roots do not have a strand really
is($location->strand, undef);
is($location->start, 83202);
is($location->end, 84996);
@sublocs = $location->sub_Location();
is(@sublocs, 2);
my $loc = shift @sublocs;
is($loc->start, 83202);
is($loc->end, 83329);
is($loc->strand, -1);
$loc = shift @sublocs;
is($loc->start, 84248);
is($loc->end, 84996);
is($loc->strand,1);
$seq = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file=> ">" .test_output_file());
$seq->verbose($verbose);
ok($seq->write_seq($as),'Fuzzy out');
## now genbank ##
$str = Bio::SeqIO->new(-format =>'genbank',
-verbose => $verbose,
-file => test_input_file('BK000016-tpa.gbk'));
$seq = $str->next_seq;
ok(defined $seq, $seq->accession_number);
ok(defined $seq->seq);
is($seq->accession_number, 'BK000016',$seq->accession_number);
is($seq->alphabet, 'dna');
is($seq->display_id, 'BK000016');
is($seq->length, 1162);
is($seq->division, 'ROD');
is($seq->get_dates, 1);
is($seq->keywords, 'Third Party Annotation; TPA');
is($seq->desc, 'TPA: Mus musculus pantothenate kinase 4 mRNA, partial cds.');
is($seq->seq_version, 1);
is($seq->feature_count, 2);
my $spec_obj = $seq->species;
is ($spec_obj->common_name, 'house mouse');
is ($spec_obj->species, 'musculus');
is ($spec_obj->genus, 'Mus');
is ($spec_obj->binomial, 'Mus musculus');
$ac = $seq->annotation;
my $reference = ($ac->get_Annotations('reference') )[0];
is ($reference->pubmed, '11479594');
is ($reference->medline, '21372465',$seq->accession_number);
# validate that what is written is what is read
my $testfile = test_output_file();
my $out = Bio::SeqIO->new(-file => ">$testfile",
-format => 'genbank');
$out->write_seq($seq);
$out->close();
$str = Bio::SeqIO->new(-format =>'genbank',
-file => $testfile);
$seq = $str->next_seq;
ok(defined $seq,'roundtrip');
ok(defined $seq->seq);
is($seq->accession_number, 'BK000016');
is($seq->alphabet, 'dna');
is($seq->display_id, 'BK000016');
is($seq->length, 1162);
is($seq->division, 'ROD');
is($seq->get_dates, 1);
is($seq->keywords, 'Third Party Annotation; TPA');
is($seq->desc, 'TPA: Mus musculus pantothenate kinase 4 mRNA, partial cds.');
is($seq->seq_version, 1);
is($seq->feature_count, 2);
$spec_obj = $seq->species;
is ($spec_obj->common_name, 'house mouse');
is ($spec_obj->species, 'musculus');
is ($spec_obj->genus, 'Mus');
is ($spec_obj->binomial, 'Mus musculus');
$ac = $seq->annotation;
$reference = ($ac->get_Annotations('reference') )[0];
is ($reference->pubmed, '11479594');
is ($reference->medline, '21372465');
# write revcomp split location
my $gb = Bio::SeqIO->new(-format => 'genbank',
# This sequence has an odd LOCUS line which sets off a warning, setting
# verbose to -1.
# The newest Ensembl seq lacks this. Maybe update? cjfields 6-5-07
-verbose => $verbose ? $verbose : -1,
-file => test_input_file('revcomp_mrna.gb'));
$seq = $gb->next_seq();
$gb = Bio::SeqIO->new(-format => 'genbank',
-file => ">$testfile");
$gb->write_seq($seq);
undef $gb;
ok(! -z $testfile, 'revcomp split location');
# bug 1925, continuation of long ORGANISM line ends up in @classification:
# ORGANISM Salmonella enterica subsp. enterica serovar Paratyphi A str. ATCC
# 9150
# Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacteriales;
# Enterobacteriaceae; Salmonella.
$gb = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('NC_006511-short.gbk'));
$seq = $gb->next_seq;
is $seq->species->common_name, undef, "Bug 1925";
is $seq->species->scientific_name, "Salmonella enterica subsp. enterica serovar Paratyphi A str. ATCC 9150";
@class = $seq->species->classification;
is $class[$#class], "Bacteria";
# WGS tests
$gb = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('O_sat.wgs'));
$seq = $gb->next_seq;
my @tests = ('wgs' => 'AAAA02000001-AAAA02050231',
'wgs_scafld' => 'CM000126-CM000137',
'wgs_scafld' => 'CH398081-CH401163');
my @wgs = map {$seq->annotation->get_Annotations(lc($_))} qw(WGS WGS_SCAFLD);
my $ct=0;
for my $wgs (@wgs) {
my ($tagname, $value) = (shift @tests, shift @tests);
is($wgs->tagname, $tagname, $tagname);
is($wgs->value, $value);
$ct++;
}
is ($ct, 3);
# make sure we can retrieve a feature with a primary tag of 'misc_difference'
$gb = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('BC000007.gbk'));
$seq = $gb->next_seq;
($cds) = grep { $_->primary_tag eq 'misc_difference' } $seq->get_SeqFeatures;
my @vals = $cds->get_tag_values('gene');
is $vals[0], 'PX19', $seq->accession_number;
# Check that the source,organism section is identical between input and output.
# - test an easy one where organism is species, then two different formats of
# subspecies, then a species with a format that used to be mistaken for
# subspecies, then a bacteria with no genus, and finally a virus with a genus.
# These tests are now somewhat out-of-date since we are moving to a Bio::Taxon-
# based system for verifying taxonomic information. Right now they just verify
# changes so are really useless; I will change them to verify common name,
# organelle, scientific name, etc.
my $outfile = test_output_file();
# output always adds a period (GenBank std), but two of these files do not use them.
foreach my $in ('BK000016-tpa.gbk', 'ay116458.gb', 'ay149291.gb', 'NC_006346.gb', 'ay007676.gb', 'dq519393.gb') {
my $infile = test_input_file($in);
$str = Bio::SeqIO->new(-format =>'genbank',
-verbose => $verbose,
-file => $infile);
$seq = $str->next_seq;
$out = Bio::SeqIO->new(-file => ">$outfile", -format => 'genbank');
$out->write_seq($seq);
$out->close();
open (IN, $infile);
my @in = <IN>;
close(IN);
open (RESULT, $outfile);
my $line = 0;
my $check = 0;
my $is = 1;
FILECHECK:
while (my $result = <RESULT>) {
if ($result =~ /^KEYWORDS/) {
$check = 1;
next;
}
if ($result =~ /^REFERENCE/) {
last FILECHECK;
}
if ($check) {
# end periods don't count (not all input files have them)
$result =~ s{\.$}{};
$in[$line] =~ s{\.$}{};
if ($result ne $in[$line]) {
$is = 0;
last;
}
}
} continue { $line++ }
close(RESULT);
ok $is, $in;
}
# NB: there should probably be full testing on all lines to ensure that output
# matches input.
# 20061117: problem with *double* colon in some annotation-dblink values
$ct = 0;
foreach my $in ('P35527.gb') {
my $infile = test_input_file($in);
$str = Bio::SeqIO->new(-format =>'genbank',
-verbose => $verbose,
-file => $infile);
$seq = $str->next_seq;
my $ac = $seq->annotation(); # Bio::AnnotationCollection
foreach my $key ($ac->get_all_annotation_keys() ) {
my @values = $ac->get_Annotations($key);
foreach my $ann (@values) {
my $value = $ann->display_text;
$ct++;
if ($key eq 'dblink') {
ok (index($value,'::') < 0); # this should never be true
ok ($value, $value); # check value is not empty
# print " ann/", sprintf('%12s ',$key), '>>>', $value , '<<<', "\n";
# print " index double colon: ",index($value ,'::'), "\n";
# check db name:
my @parts = split(/:/,$value);
if ( $parts[0] =~ /^(?:
# not an exhaustive list of databases;
# just the db's referenced in P35527.gb:
swissprot | GenBank | GenPept | HSSP| IntAct | Ensembl | KEGG | HGNC | MIM | ArrayExpress
| GO | InterPro | Pfam| PRINTS | PROSITE
)$/x )
{
ok 1;
}
else {
ok 0;
}
ok ( $parts[1], "$parts[0]" );
}
}
}
}
is($ct, 46);
# bug 2195
$str = Bio::SeqIO->new(-format =>'genbank',
-verbose => $verbose,
-file => test_input_file('AF305198.gb')
);
$species = $str->next_seq->species;
is($species->scientific_name, 'Virginia creeper phytoplasma', 'Bug 2195');
is(join(', ',$species->classification), 'Virginia creeper phytoplasma, '.
'16SrV (Elm yellows group), Candidatus Phytoplasma, '.
'Acholeplasmataceae, Acholeplasmatales, Mollicutes, '.
'Firmicutes, Bacteria', 'Bug 2195');
# bug 2569, PROJECT line support, read and write, round-tripping
$str = Bio::SeqIO->new(-format =>'genbank',
-verbose => $verbose,
-file => test_input_file('NC_008536.gb'));
$seq = $str->next_seq;
my $project = ($seq->annotation->get_Annotations('project'))[0];
isa_ok($project, 'Bio::Annotation::SimpleValue');
if ($project) {
is($project->value, 'GenomeProject:12638');
} else {
ok(0, "PROJECT not parsed");
}
$outfile = test_output_file();
$gb = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file=> ">$outfile");
$gb->write_seq($seq);
$str = Bio::SeqIO->new(-format =>'genbank',
-verbose => $verbose,
-file => $outfile);
$seq = $str->next_seq;
$project = ($seq->annotation->get_Annotations('project'))[0];
isa_ok($project, 'Bio::Annotation::SimpleValue');
if ($project) {
is($project->value, 'GenomeProject:12638');
} else {
ok(0, "Roundtrip test failed");
}
# test for swissprot/UniProt/UniProtKB DBSOURCE line (Bug : RT 44536)
$ast = Bio::SeqIO->new(-format => 'genbank',
-verbose => $verbose,
-file => test_input_file('P39765.gb'));
$as = $ast->next_seq();
is $as->molecule, 'PRT',$as->accession_number;
is $as->division, 'BCT',$as->accession_number;
is join(',',$as->get_dates), '03-MAR-2009',$as->accession_number;
is $as->alphabet, 'protein';
# Though older GenBank releases indicate SOURCE contains only the common name,
# this is no longer true. In general, this line will contain an abbreviated
# form of the full organism name (but may contain the full length name),
# as well as the optional common name and organelle. There is no get/set
# for the abbreviated name but it is accessible via name()
ok defined($as->species->name('abbreviated')->[0]);
is $as->species->name('abbreviated')->[0], 'Bacillus subtilis';
is($as->primary_id, 20141743);
$ac = $as->annotation;
ok defined $ac;
@dblinks = $ac->get_Annotations('dblink');
is(scalar @dblinks,31);
is($dblinks[0]->database, 'UniProtKB');
is($dblinks[0]->primary_id, 'PYRR_BACSU');
is($dblinks[0]->version, undef);
is($dblinks[0]->display_text, 'UniProtKB:PYRR_BACSU','operator overloading in AnnotationI is deprecated');
#bug 2982 embl/genbank contig handling
$ast = Bio::SeqIO->new( -file => test_input_file('bug2982.gb'),
-format => 'genbank' );
$seq = $ast->next_seq;
ok my @ctg = $seq->annotation->get_Annotations('contig');
like $ctg[0]->value, qr/join\(.*?gap.*?complement/;
# write_seq() and FTHelper duplicate specific tags, need to check a round-trip
$ast = Bio::SeqIO->new(-format => 'genbank' ,
-verbose => $verbose,
-file => test_input_file('singlescore.gbk'));
$as = $ast->next_seq();
($cds) = grep { $_->primary_tag eq 'CDS' } $as->get_SeqFeatures();
my @notes = $cds->get_tag_values('note');
is(scalar @notes, 2);
$testfile = test_output_file();
$out = Bio::SeqIO->new(-file => ">$testfile",
-format => 'genbank');
$out->write_seq($as);
$out->close();
$ast = Bio::SeqIO->new(-format => 'genbank' ,
-verbose => $verbose,
-file => $testfile );
$as = $ast->next_seq;
($cds) = grep { $_->primary_tag eq 'CDS' } $as->get_SeqFeatures();
@notes = $cds->get_tag_values('note');
is(scalar @notes, 2);
#bug 3375
my $in = Bio::SeqIO->new(-format => 'genbank',
-file => test_input_file('NC_002058_multDBLINK_bug3375.gb'));
$seq = $in->next_seq(); # should not throw a warning now
@dblinks = $seq->annotation->get_Annotations('dblink'); # contains 5 dblink references
# testing DBLINK BioProject: PRJNA15288
is($dblinks[0]->database, 'BioProject', 'bug3375 database is BioProject');
is($dblinks[0]->primary_id, 'PRJNA15288', 'bug3375 primary_id is PRJNA15288');
# testing DBLINK Project:100,200,300
is($dblinks[3]->database, 'Project');
is($dblinks[3]->primary_id, '300');
# testing DBLINK NC_002058.3
is($dblinks[4]->database, 'GenBank');
is($dblinks[4]->primary_id, 'NC_002058');
is($dblinks[4]->version, '3');
# long labels handled
{
# Create sequence with feature with a long label qualifier
my $seq=Bio::Seq->new(-seq => 'actg',
-id => 'abacab');
my $feature=Bio::SeqFeature::Generic->new(-primary=>'CDS', -start=>1, -end=>4);
my $label='1 2 3 4 5 6 7 8 9 a b c d e f g h i j k l m n o p q r';
$feature->add_tag_value(label=>$label);
$seq->add_SeqFeature($feature);
# Write genbank
my $string;
open(my $str_fh, '>', \$string) || skip("Can't open string, skipping", 2);
my $out=Bio::SeqIO->new(-format=>'genbank', -fh => $str_fh);
$out->write_seq($seq);
# Read genbank
my $in=Bio::SeqIO->new(-format=>'genbank', -string => $string);
my $genbank=$in->next_seq;
my ($read_feature)=$genbank->get_SeqFeatures;
my ($read_label)=$read_feature->get_tag_values('label');
is($read_label, $label, 'Label is the same');
}