line |
stmt |
bran |
cond |
sub |
pod |
time |
code |
1
|
|
|
|
|
|
|
# |
2
|
|
|
|
|
|
|
# BioPerl module for Bio::SeqFeature::Gene::Intron |
3
|
|
|
|
|
|
|
# |
4
|
|
|
|
|
|
|
# Please direct questions and support issues to |
5
|
|
|
|
|
|
|
# |
6
|
|
|
|
|
|
|
# Cared for by David Block |
7
|
|
|
|
|
|
|
# |
8
|
|
|
|
|
|
|
# Copyright David Block |
9
|
|
|
|
|
|
|
# |
10
|
|
|
|
|
|
|
# You may distribute this module under the same terms as perl itself |
11
|
|
|
|
|
|
|
|
12
|
|
|
|
|
|
|
# POD documentation - main docs before the code |
13
|
|
|
|
|
|
|
|
14
|
|
|
|
|
|
|
=head1 NAME |
15
|
|
|
|
|
|
|
|
16
|
|
|
|
|
|
|
Bio::SeqFeature::Gene::Intron - An intron feature |
17
|
|
|
|
|
|
|
|
18
|
|
|
|
|
|
|
=head1 SYNOPSIS |
19
|
|
|
|
|
|
|
|
20
|
|
|
|
|
|
|
Give standard usage here |
21
|
|
|
|
|
|
|
|
22
|
|
|
|
|
|
|
=head1 DESCRIPTION |
23
|
|
|
|
|
|
|
|
24
|
|
|
|
|
|
|
Describe the object here |
25
|
|
|
|
|
|
|
|
26
|
|
|
|
|
|
|
=head1 FEEDBACK |
27
|
|
|
|
|
|
|
|
28
|
|
|
|
|
|
|
=head2 Mailing Lists |
29
|
|
|
|
|
|
|
|
30
|
|
|
|
|
|
|
User feedback is an integral part of the evolution of this and other |
31
|
|
|
|
|
|
|
Bioperl modules. Send your comments and suggestions preferably to |
32
|
|
|
|
|
|
|
the Bioperl mailing list. Your participation is much appreciated. |
33
|
|
|
|
|
|
|
|
34
|
|
|
|
|
|
|
bioperl-l@bioperl.org - General discussion |
35
|
|
|
|
|
|
|
http://bioperl.org/wiki/Mailing_lists - About the mailing lists |
36
|
|
|
|
|
|
|
|
37
|
|
|
|
|
|
|
=head2 Support |
38
|
|
|
|
|
|
|
|
39
|
|
|
|
|
|
|
Please direct usage questions or support issues to the mailing list: |
40
|
|
|
|
|
|
|
|
41
|
|
|
|
|
|
|
I |
42
|
|
|
|
|
|
|
|
43
|
|
|
|
|
|
|
rather than to the module maintainer directly. Many experienced and |
44
|
|
|
|
|
|
|
reponsive experts will be able look at the problem and quickly |
45
|
|
|
|
|
|
|
address it. Please include a thorough description of the problem |
46
|
|
|
|
|
|
|
with code and data examples if at all possible. |
47
|
|
|
|
|
|
|
|
48
|
|
|
|
|
|
|
=head2 Reporting Bugs |
49
|
|
|
|
|
|
|
|
50
|
|
|
|
|
|
|
Report bugs to the Bioperl bug tracking system to help us keep track |
51
|
|
|
|
|
|
|
of the bugs and their resolution. Bug reports can be submitted via the |
52
|
|
|
|
|
|
|
web: |
53
|
|
|
|
|
|
|
|
54
|
|
|
|
|
|
|
https://github.com/bioperl/bioperl-live/issues |
55
|
|
|
|
|
|
|
|
56
|
|
|
|
|
|
|
=head1 AUTHOR - David Block |
57
|
|
|
|
|
|
|
|
58
|
|
|
|
|
|
|
Email dblock@gene.pbi.nrc.ca |
59
|
|
|
|
|
|
|
|
60
|
|
|
|
|
|
|
=head1 APPENDIX |
61
|
|
|
|
|
|
|
|
62
|
|
|
|
|
|
|
The rest of the documentation details each of the object methods. |
63
|
|
|
|
|
|
|
Internal methods are usually preceded with a _ |
64
|
|
|
|
|
|
|
|
65
|
|
|
|
|
|
|
=cut |
66
|
|
|
|
|
|
|
|
67
|
|
|
|
|
|
|
|
68
|
|
|
|
|
|
|
# Let the code begin... |
69
|
|
|
|
|
|
|
|
70
|
|
|
|
|
|
|
|
71
|
|
|
|
|
|
|
package Bio::SeqFeature::Gene::Intron; |
72
|
8
|
|
|
8
|
|
27
|
use strict; |
|
8
|
|
|
|
|
8
|
|
|
8
|
|
|
|
|
184
|
|
73
|
|
|
|
|
|
|
|
74
|
8
|
|
|
8
|
|
24
|
use Bio::SeqFeature::Gene::Exon; |
|
8
|
|
|
|
|
9
|
|
|
8
|
|
|
|
|
202
|
|
75
|
|
|
|
|
|
|
|
76
|
8
|
|
|
8
|
|
24
|
use base qw(Bio::SeqFeature::Gene::NC_Feature); |
|
8
|
|
|
|
|
8
|
|
|
8
|
|
|
|
|
5485
|
|
77
|
|
|
|
|
|
|
|
78
|
|
|
|
|
|
|
sub new { |
79
|
3
|
|
|
3
|
1
|
5
|
my($class,@args) = @_; |
80
|
|
|
|
|
|
|
|
81
|
|
|
|
|
|
|
# introns are non-coding by default |
82
|
3
|
50
|
|
|
|
4
|
if(! grep { lc($_) eq '-is_coding'; } @args) { |
|
6
|
|
|
|
|
12
|
|
83
|
3
|
|
|
|
|
3
|
push(@args, '-is_coding', 0); |
84
|
|
|
|
|
|
|
} |
85
|
3
|
|
|
|
|
10
|
my $self = $class->SUPER::new(@args); |
86
|
|
|
|
|
|
|
|
87
|
3
|
|
|
|
|
8
|
my ($primary, $prim) = |
88
|
|
|
|
|
|
|
$self->_rearrange([qw(PRIMARY PRIMARY_TAG)],@args); |
89
|
3
|
50
|
33
|
|
|
8
|
$self->primary_tag('intron') unless $primary || $prim; |
90
|
|
|
|
|
|
|
|
91
|
3
|
|
|
|
|
6
|
return $self; |
92
|
|
|
|
|
|
|
} |
93
|
|
|
|
|
|
|
|
94
|
|
|
|
|
|
|
=head2 upstream_Exon |
95
|
|
|
|
|
|
|
|
96
|
|
|
|
|
|
|
Title : upstream_Exon |
97
|
|
|
|
|
|
|
Usage : $intron->upstream_Exon() |
98
|
|
|
|
|
|
|
Function: exon upstream of the intron |
99
|
|
|
|
|
|
|
Returns : Bio::EnsEMBL::Exon |
100
|
|
|
|
|
|
|
Args : |
101
|
|
|
|
|
|
|
|
102
|
|
|
|
|
|
|
=cut |
103
|
|
|
|
|
|
|
|
104
|
|
|
|
|
|
|
sub upstream_Exon { |
105
|
6
|
|
|
6
|
1
|
6
|
my( $self, $exon ) = @_; |
106
|
|
|
|
|
|
|
|
107
|
6
|
100
|
|
|
|
8
|
if ($exon) { |
108
|
3
|
|
|
|
|
3
|
$self->{'_intron_location'} = undef; |
109
|
3
|
50
|
|
|
|
8
|
$self->throw("'$exon' is not a Bio::SeqFeature::Gene::ExonI") |
110
|
|
|
|
|
|
|
unless $exon->isa('Bio::SeqFeature::Gene::ExonI'); |
111
|
3
|
|
|
|
|
7
|
$self->{'_upstream_exon'} = $exon; |
112
|
|
|
|
|
|
|
} |
113
|
6
|
|
|
|
|
8
|
return $self->{'_upstream_exon'}; |
114
|
|
|
|
|
|
|
} |
115
|
|
|
|
|
|
|
|
116
|
|
|
|
|
|
|
|
117
|
|
|
|
|
|
|
=head2 downstream_Exon |
118
|
|
|
|
|
|
|
|
119
|
|
|
|
|
|
|
Title : downstream_Exon |
120
|
|
|
|
|
|
|
Usage : $intron->downstream_Exon() |
121
|
|
|
|
|
|
|
Function: exon downstream of the intron |
122
|
|
|
|
|
|
|
Returns : Bio::EnsEMBL::Exon |
123
|
|
|
|
|
|
|
Args : |
124
|
|
|
|
|
|
|
|
125
|
|
|
|
|
|
|
=cut |
126
|
|
|
|
|
|
|
|
127
|
|
|
|
|
|
|
sub downstream_Exon { |
128
|
6
|
|
|
6
|
1
|
6
|
my( $self, $exon ) = @_; |
129
|
|
|
|
|
|
|
|
130
|
6
|
100
|
|
|
|
8
|
if ($exon) { |
131
|
3
|
|
|
|
|
3
|
$self->{'_intron_location'} = undef; |
132
|
3
|
50
|
|
|
|
7
|
$self->throw("'$exon' is not a Bio::SeqFeature::Gene::ExonI") |
133
|
|
|
|
|
|
|
unless $exon->isa('Bio::SeqFeature::Gene::ExonI'); |
134
|
3
|
|
|
|
|
8
|
$self->{'_downstream_exon'} = $exon; |
135
|
|
|
|
|
|
|
} |
136
|
6
|
|
|
|
|
5
|
return $self->{'_downstream_exon'}; |
137
|
|
|
|
|
|
|
} |
138
|
|
|
|
|
|
|
|
139
|
|
|
|
|
|
|
=head2 phase |
140
|
|
|
|
|
|
|
|
141
|
|
|
|
|
|
|
Title : phase |
142
|
|
|
|
|
|
|
Usage : $intron->phase() |
143
|
|
|
|
|
|
|
Function: returns the phase of the intron(where it interrupts the codon) |
144
|
|
|
|
|
|
|
Returns : int(0,1,2) |
145
|
|
|
|
|
|
|
Args : |
146
|
|
|
|
|
|
|
|
147
|
|
|
|
|
|
|
=cut |
148
|
|
|
|
|
|
|
|
149
|
|
|
|
|
|
|
sub phase { |
150
|
0
|
|
|
0
|
1
|
0
|
my ($self) = @_; |
151
|
0
|
|
|
|
|
0
|
return $self->downstream_Exon->phase; |
152
|
|
|
|
|
|
|
} |
153
|
|
|
|
|
|
|
|
154
|
|
|
|
|
|
|
|
155
|
|
|
|
|
|
|
=head2 acceptor_splice_site |
156
|
|
|
|
|
|
|
|
157
|
|
|
|
|
|
|
Title : acceptor_splice_site |
158
|
|
|
|
|
|
|
Usage : $intron->acceptor_splice_site(21,3) |
159
|
|
|
|
|
|
|
Function: returns the sequence corresponding to the |
160
|
|
|
|
|
|
|
consensus acceptor splice site. If start and |
161
|
|
|
|
|
|
|
end are provided, it will number of base pairs |
162
|
|
|
|
|
|
|
left and right of the canonical AG. Here 21 means |
163
|
|
|
|
|
|
|
21 bp into intron and 3 means 3 bp into the exon. |
164
|
|
|
|
|
|
|
--Intron--21----|AG|-3-----Exon |
165
|
|
|
|
|
|
|
Defaults to 21,3 |
166
|
|
|
|
|
|
|
|
167
|
|
|
|
|
|
|
Returns : Bio::Seq |
168
|
|
|
|
|
|
|
Args : start and end |
169
|
|
|
|
|
|
|
|
170
|
|
|
|
|
|
|
=cut |
171
|
|
|
|
|
|
|
|
172
|
|
|
|
|
|
|
sub acceptor_splice_site { |
173
|
0
|
|
|
0
|
1
|
0
|
my ($self,$ss_start,$ss_end) = @_; |
174
|
0
|
0
|
|
|
|
0
|
$ss_start = 21 unless defined $ss_start; |
175
|
0
|
0
|
|
|
|
0
|
$ss_end = 3 unless defined $ss_end; |
176
|
0
|
0
|
|
|
|
0
|
if($self->strand < 0){ |
177
|
0
|
|
|
|
|
0
|
my $tmp= $ss_start; |
178
|
0
|
|
|
|
|
0
|
$ss_start = $ss_end; |
179
|
0
|
|
|
|
|
0
|
$ss_end = $tmp; |
180
|
|
|
|
|
|
|
} |
181
|
0
|
|
|
|
|
0
|
my $intron_end= $self->location->end; |
182
|
0
|
|
|
|
|
0
|
my $down_exon = $self->downstream_Exon; |
183
|
0
|
|
|
|
|
0
|
my $acceptor; |
184
|
0
|
0
|
|
|
|
0
|
if($self->strand < 0){ |
185
|
0
|
0
|
|
|
|
0
|
$ss_start= $ss_start > $down_exon->length ? $down_exon->length: $ss_start; |
186
|
0
|
0
|
|
|
|
0
|
$ss_end= $ss_end > $self->length-2 ? $self->length-2 : $ss_end; |
187
|
0
|
|
|
|
|
0
|
$acceptor = Bio::SeqFeature::Generic->new(-start=>$self->start - ($ss_start) , |
188
|
|
|
|
|
|
|
-end=>$self->start + ($ss_end+1), |
189
|
|
|
|
|
|
|
-strand=>$self->strand, |
190
|
|
|
|
|
|
|
-primary_tag=>"donor splice site"); |
191
|
|
|
|
|
|
|
} |
192
|
|
|
|
|
|
|
else { |
193
|
0
|
0
|
|
|
|
0
|
$ss_start = $ss_start > $self->length-2 ? $self->length-2 : $ss_start; |
194
|
0
|
0
|
|
|
|
0
|
$ss_end = $ss_end > $down_exon->length ? $down_exon->length : $ss_end; |
195
|
|
|
|
|
|
|
|
196
|
|
|
|
|
|
|
|
197
|
0
|
|
|
|
|
0
|
$acceptor = Bio::SeqFeature::Generic->new(-start=>$self->end - ($ss_start + 1), |
198
|
|
|
|
|
|
|
-end=>$self->end + $ss_end, |
199
|
|
|
|
|
|
|
-strand=>$self->strand, |
200
|
|
|
|
|
|
|
-primary_tag=>"donor splice site"); |
201
|
|
|
|
|
|
|
} |
202
|
0
|
|
|
|
|
0
|
$acceptor->attach_seq($self->entire_seq); |
203
|
|
|
|
|
|
|
|
204
|
0
|
|
|
|
|
0
|
return $acceptor; |
205
|
|
|
|
|
|
|
|
206
|
|
|
|
|
|
|
} |
207
|
|
|
|
|
|
|
|
208
|
|
|
|
|
|
|
=head2 donor_splice_site |
209
|
|
|
|
|
|
|
|
210
|
|
|
|
|
|
|
Title : donor_splice_site |
211
|
|
|
|
|
|
|
Usage : $intron->donor_splice_site(3,6) |
212
|
|
|
|
|
|
|
Function: returns the sequence corresponding to the |
213
|
|
|
|
|
|
|
consensus donor splice site. If start and |
214
|
|
|
|
|
|
|
end are provided, it will number of base pairs |
215
|
|
|
|
|
|
|
left and right of the canonical GT. Here 3 means |
216
|
|
|
|
|
|
|
3 bp into exon and 6 means 6 bp into the intron. |
217
|
|
|
|
|
|
|
--Exon-3--|GT|-6----Intron- |
218
|
|
|
|
|
|
|
Defaults to 3,6 |
219
|
|
|
|
|
|
|
|
220
|
|
|
|
|
|
|
Returns : Bio::Seq |
221
|
|
|
|
|
|
|
Args : start and end |
222
|
|
|
|
|
|
|
|
223
|
|
|
|
|
|
|
=cut |
224
|
|
|
|
|
|
|
|
225
|
|
|
|
|
|
|
sub donor_splice_site { |
226
|
0
|
|
|
0
|
1
|
0
|
my ($self,$ss_start,$ss_end) = @_; |
227
|
0
|
0
|
|
|
|
0
|
$ss_start = 3 unless defined $ss_start; |
228
|
0
|
0
|
|
|
|
0
|
$ss_end = 10 unless defined $ss_end; |
229
|
0
|
0
|
|
|
|
0
|
if($self->strand < 0){ |
230
|
0
|
|
|
|
|
0
|
my $tmp= $ss_start; |
231
|
0
|
|
|
|
|
0
|
$ss_start = $ss_end; |
232
|
0
|
|
|
|
|
0
|
$ss_end = $tmp; |
233
|
|
|
|
|
|
|
} |
234
|
0
|
|
|
|
|
0
|
my $up_exon = $self->upstream_Exon; |
235
|
0
|
|
|
|
|
0
|
my $donor; |
236
|
0
|
0
|
|
|
|
0
|
if($self->strand < 0){ |
237
|
0
|
0
|
|
|
|
0
|
$ss_end = $ss_end > $up_exon->length ? $up_exon->length : $ss_end; |
238
|
0
|
0
|
|
|
|
0
|
$ss_start = $ss_start> $self->length -2 ? $self->length -2 : $ss_start; |
239
|
0
|
|
|
|
|
0
|
$donor = Bio::SeqFeature::Generic->new(-start=>$self->end - ($ss_start+1), |
240
|
|
|
|
|
|
|
-end => $self->end + ($ss_end), |
241
|
|
|
|
|
|
|
-strand=>$self->strand, |
242
|
|
|
|
|
|
|
-primary_tag=>"acceptor splice site"); |
243
|
|
|
|
|
|
|
} |
244
|
|
|
|
|
|
|
else { |
245
|
0
|
0
|
|
|
|
0
|
$ss_start = $ss_start > $up_exon->length ? $up_exon->length : $ss_start; |
246
|
0
|
0
|
|
|
|
0
|
$ss_end = $ss_end > $self->length -2 ? $self->length -2 : $ss_end; |
247
|
0
|
|
|
|
|
0
|
$donor = Bio::SeqFeature::Generic->new(-start=>$self->start - $ss_start, |
248
|
|
|
|
|
|
|
-end => $self->start +($ss_end+1), |
249
|
|
|
|
|
|
|
-strand=>$self->strand, |
250
|
|
|
|
|
|
|
-primary_tag=>"acceptor splice site"); |
251
|
|
|
|
|
|
|
} |
252
|
0
|
|
|
|
|
0
|
$donor->attach_seq($self->entire_seq); |
253
|
0
|
|
|
|
|
0
|
return $donor; |
254
|
|
|
|
|
|
|
} |
255
|
|
|
|
|
|
|
|
256
|
|
|
|
|
|
|
sub location { |
257
|
9
|
|
|
9
|
1
|
8
|
my( $self ) = @_; |
258
|
|
|
|
|
|
|
|
259
|
9
|
100
|
|
|
|
18
|
unless ($self->{'_intron_location'}) { |
260
|
3
|
|
|
|
|
8
|
my $loc = Bio::Location::Simple->new; |
261
|
|
|
|
|
|
|
|
262
|
3
|
|
|
|
|
3
|
my $up_exon = $self->upstream_Exon; |
263
|
3
|
|
|
|
|
3
|
my $down_exon = $self->downstream_Exon; |
264
|
|
|
|
|
|
|
|
265
|
|
|
|
|
|
|
# Get the PrimarySeqs attached to both and check it is the same sequence |
266
|
3
|
|
|
|
|
7
|
my $up_seq = $up_exon ->entire_seq; |
267
|
3
|
|
|
|
|
4
|
my $down_seq = $down_exon->entire_seq; |
268
|
3
|
50
|
|
|
|
6
|
unless (ref($up_seq) eq ref($down_seq) ) { |
269
|
0
|
|
|
|
|
0
|
$self->throw("upstream and downstream exons are attached to different sequences\n'$up_seq' and '$down_seq'"); |
270
|
|
|
|
|
|
|
} |
271
|
|
|
|
|
|
|
|
272
|
|
|
|
|
|
|
# Check that the exons are on the same strand. (Do I need to bother?) |
273
|
3
|
|
|
|
|
5
|
my $up_strand = $up_exon ->strand; |
274
|
3
|
|
|
|
|
4
|
my $down_strand = $down_exon->strand; |
275
|
3
|
50
|
|
|
|
5
|
unless ($up_strand == $down_strand) { |
276
|
0
|
|
|
|
|
0
|
$self->throw("upstream and downstream exons are on different strands " |
277
|
|
|
|
|
|
|
. "('$up_strand' and '$down_strand')"); |
278
|
|
|
|
|
|
|
} |
279
|
3
|
|
|
|
|
4
|
$loc->strand($up_strand); |
280
|
|
|
|
|
|
|
|
281
|
|
|
|
|
|
|
# $exon_end is the end of the exon which is 5' of the intron on the genomic sequence. |
282
|
|
|
|
|
|
|
# $exon_start is the start of the exon which is 3' of the intron on the genomic sequence. |
283
|
3
|
|
|
|
|
3
|
my( $exon_end, $exon_start ); |
284
|
3
|
50
|
|
|
|
5
|
if ($up_strand == 1) { |
285
|
3
|
|
|
|
|
10
|
$exon_end = $up_exon ->end; |
286
|
3
|
|
|
|
|
9
|
$exon_start = $down_exon->start; |
287
|
|
|
|
|
|
|
} else { |
288
|
0
|
|
|
|
|
0
|
$exon_end = $down_exon->end; |
289
|
0
|
|
|
|
|
0
|
$exon_start = $up_exon ->start; |
290
|
|
|
|
|
|
|
} |
291
|
3
|
50
|
|
|
|
11
|
unless ($exon_end < $exon_start) { |
292
|
0
|
|
|
|
|
0
|
$self->throw("Intron gap begins after '$exon_end' and ends before '$exon_start'"); |
293
|
|
|
|
|
|
|
} |
294
|
3
|
|
|
|
|
10
|
$loc->start($exon_end + 1); |
295
|
3
|
|
|
|
|
6
|
$loc->end ($exon_start - 1); |
296
|
|
|
|
|
|
|
|
297
|
|
|
|
|
|
|
# Attach the sequence and location objects to the intron |
298
|
3
|
|
|
|
|
4
|
$self->{'_intron_location'} = $loc; |
299
|
|
|
|
|
|
|
|
300
|
|
|
|
|
|
|
} |
301
|
9
|
|
|
|
|
17
|
return $self->{'_intron_location'}; |
302
|
|
|
|
|
|
|
} |
303
|
|
|
|
|
|
|
1; |