line |
stmt |
bran |
cond |
sub |
pod |
time |
code |
1
|
|
|
|
|
|
|
# |
2
|
|
|
|
|
|
|
# GeneDesign module for sequence segmentation |
3
|
|
|
|
|
|
|
# |
4
|
|
|
|
|
|
|
|
5
|
|
|
|
|
|
|
=head1 NAME |
6
|
|
|
|
|
|
|
|
7
|
|
|
|
|
|
|
Bio::GeneDesign::Oligo |
8
|
|
|
|
|
|
|
|
9
|
|
|
|
|
|
|
=head1 VERSION |
10
|
|
|
|
|
|
|
|
11
|
|
|
|
|
|
|
Version 5.52 |
12
|
|
|
|
|
|
|
|
13
|
|
|
|
|
|
|
=head1 DESCRIPTION |
14
|
|
|
|
|
|
|
|
15
|
|
|
|
|
|
|
|
16
|
|
|
|
|
|
|
=head1 AUTHOR |
17
|
|
|
|
|
|
|
|
18
|
|
|
|
|
|
|
Sarah Richardson <SMRichardson@lbl.gov>. |
19
|
|
|
|
|
|
|
|
20
|
|
|
|
|
|
|
=cut |
21
|
|
|
|
|
|
|
|
22
|
|
|
|
|
|
|
package Bio::GeneDesign::Oligo; |
23
|
|
|
|
|
|
|
require Exporter; |
24
|
|
|
|
|
|
|
|
25
|
11
|
|
|
11
|
|
176
|
use Bio::GeneDesign::Basic qw(:GD); |
|
11
|
|
|
|
|
20
|
|
|
11
|
|
|
|
|
3003
|
|
26
|
11
|
|
|
11
|
|
70
|
use strict; |
|
11
|
|
|
|
|
22
|
|
|
11
|
|
|
|
|
474
|
|
27
|
11
|
|
|
11
|
|
64
|
use warnings; |
|
11
|
|
|
|
|
29
|
|
|
11
|
|
|
|
|
709
|
|
28
|
|
|
|
|
|
|
|
29
|
|
|
|
|
|
|
our $VERSION = 5.52; |
30
|
|
|
|
|
|
|
|
31
|
11
|
|
|
11
|
|
65
|
use base qw(Exporter); |
|
11
|
|
|
|
|
20
|
|
|
11
|
|
|
|
|
7050
|
|
32
|
|
|
|
|
|
|
our @EXPORT_OK = qw( |
33
|
|
|
|
|
|
|
_make_amplification_primers |
34
|
|
|
|
|
|
|
_filter_homopolymer |
35
|
|
|
|
|
|
|
_check_for_homopolymer |
36
|
|
|
|
|
|
|
$VERSION |
37
|
|
|
|
|
|
|
); |
38
|
|
|
|
|
|
|
our %EXPORT_TAGS = (GD => \@EXPORT_OK); |
39
|
|
|
|
|
|
|
|
40
|
|
|
|
|
|
|
=head1 Functions |
41
|
|
|
|
|
|
|
|
42
|
|
|
|
|
|
|
=head2 _filter_homopolymer() |
43
|
|
|
|
|
|
|
|
44
|
|
|
|
|
|
|
=cut |
45
|
|
|
|
|
|
|
|
46
|
|
|
|
|
|
|
sub _filter_homopolymer |
47
|
|
|
|
|
|
|
{ |
48
|
0
|
|
|
0
|
|
|
my ($seqarr, $length) = @_; |
49
|
0
|
|
|
|
|
|
my @newarr = (); |
50
|
0
|
|
|
|
|
|
foreach my $seq (@{$seqarr}) |
|
0
|
|
|
|
|
|
|
51
|
|
|
|
|
|
|
{ |
52
|
0
|
0
|
|
|
|
|
push @newarr, $seq if (! _check_for_homopolymer($seq, $length)); |
53
|
|
|
|
|
|
|
} |
54
|
0
|
|
|
|
|
|
return \@newarr; |
55
|
|
|
|
|
|
|
} |
56
|
|
|
|
|
|
|
|
57
|
|
|
|
|
|
|
=head2 _check_for_homopolymer() |
58
|
|
|
|
|
|
|
|
59
|
|
|
|
|
|
|
=cut |
60
|
|
|
|
|
|
|
|
61
|
|
|
|
|
|
|
sub _check_for_homopolymer |
62
|
|
|
|
|
|
|
{ |
63
|
0
|
|
|
0
|
|
|
my ($seq, $length) = @_; |
64
|
0
|
|
0
|
|
|
|
$length = $length || 5; |
65
|
0
|
0
|
|
|
|
|
return 1 if ($length <= 1); |
66
|
0
|
0
|
|
|
|
|
return 1 if $seq =~ m{A{$length}|T{$length}|C{$length}|G{$length}}msxi; |
67
|
0
|
|
|
|
|
|
return 0; |
68
|
|
|
|
|
|
|
} |
69
|
|
|
|
|
|
|
|
70
|
|
|
|
|
|
|
=head2 _make_amplification_primers() |
71
|
|
|
|
|
|
|
|
72
|
|
|
|
|
|
|
=cut |
73
|
|
|
|
|
|
|
|
74
|
|
|
|
|
|
|
sub _make_amplification_primers |
75
|
|
|
|
|
|
|
{ |
76
|
0
|
|
|
0
|
|
|
my ($sequence, $temperature) = @_; |
77
|
|
|
|
|
|
|
|
78
|
0
|
|
|
|
|
|
my $left_length = 5; |
79
|
0
|
|
|
|
|
|
my $lprimer = substr($sequence, 0, $left_length); |
80
|
0
|
|
|
|
|
|
while (_melt($lprimer) < $temperature) |
81
|
|
|
|
|
|
|
{ |
82
|
0
|
|
|
|
|
|
$left_length++; |
83
|
0
|
0
|
|
|
|
|
last if ($left_length > 45); |
84
|
0
|
|
|
|
|
|
$lprimer = substr($sequence, 0, $left_length) |
85
|
|
|
|
|
|
|
} |
86
|
|
|
|
|
|
|
|
87
|
0
|
|
|
|
|
|
my $right_end = length($sequence); |
88
|
0
|
|
|
|
|
|
my $right_length = 5; |
89
|
0
|
|
|
|
|
|
my $rprimer = substr($sequence, $right_end - $right_length, $right_length); |
90
|
0
|
|
|
|
|
|
while (_melt($rprimer) < $temperature) |
91
|
|
|
|
|
|
|
{ |
92
|
0
|
|
|
|
|
|
$right_length++; |
93
|
0
|
0
|
|
|
|
|
last if ($right_length > 45); |
94
|
0
|
|
|
|
|
|
$rprimer = substr($sequence, $right_end - $right_length, $right_length); |
95
|
|
|
|
|
|
|
} |
96
|
0
|
|
|
|
|
|
$rprimer = _complement($rprimer, 1); |
97
|
|
|
|
|
|
|
|
98
|
0
|
|
|
|
|
|
return ($lprimer, $rprimer); |
99
|
|
|
|
|
|
|
} |
100
|
|
|
|
|
|
|
|
101
|
|
|
|
|
|
|
1; |
102
|
|
|
|
|
|
|
|
103
|
|
|
|
|
|
|
__END__ |
104
|
|
|
|
|
|
|
|
105
|
|
|
|
|
|
|
=head1 COPYRIGHT AND LICENSE |
106
|
|
|
|
|
|
|
|
107
|
|
|
|
|
|
|
Copyright (c) 2013, GeneDesign developers |
108
|
|
|
|
|
|
|
All rights reserved. |
109
|
|
|
|
|
|
|
|
110
|
|
|
|
|
|
|
Redistribution and use in source and binary forms, with or without modification, |
111
|
|
|
|
|
|
|
are permitted provided that the following conditions are met: |
112
|
|
|
|
|
|
|
|
113
|
|
|
|
|
|
|
* Redistributions of source code must retain the above copyright notice, this |
114
|
|
|
|
|
|
|
list of conditions and the following disclaimer. |
115
|
|
|
|
|
|
|
|
116
|
|
|
|
|
|
|
* Redistributions in binary form must reproduce the above copyright notice, this |
117
|
|
|
|
|
|
|
list of conditions and the following disclaimer in the documentation and/or |
118
|
|
|
|
|
|
|
other materials provided with the distribution. |
119
|
|
|
|
|
|
|
|
120
|
|
|
|
|
|
|
* The names of Johns Hopkins, the Joint Genome Institute, the Lawrence Berkeley |
121
|
|
|
|
|
|
|
National Laboratory, the Department of Energy, and the GeneDesign developers may |
122
|
|
|
|
|
|
|
not be used to endorse or promote products derived from this software without |
123
|
|
|
|
|
|
|
specific prior written permission. |
124
|
|
|
|
|
|
|
|
125
|
|
|
|
|
|
|
THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS "AS IS" AND |
126
|
|
|
|
|
|
|
ANY EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED |
127
|
|
|
|
|
|
|
WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE |
128
|
|
|
|
|
|
|
DISCLAIMED. IN NO EVENT SHALL THE DEVELOPERS BE LIABLE FOR ANY DIRECT, INDIRECT, |
129
|
|
|
|
|
|
|
INCIDENTAL, SPECIAL, EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT |
130
|
|
|
|
|
|
|
LIMITED TO, PROCUREMENT OF SUBSTITUTE GOODS OR SERVICES; LOSS OF USE, DATA, OR |
131
|
|
|
|
|
|
|
PROFITS; OR BUSINESS INTERRUPTION) HOWEVER CAUSED AND ON ANY THEORY OF |
132
|
|
|
|
|
|
|
LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR TORT (INCLUDING NEGLIGENCE |
133
|
|
|
|
|
|
|
OR OTHERWISE) ARISING IN ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF |
134
|
|
|
|
|
|
|
ADVISED OF THE POSSIBILITY OF SUCH DAMAGE. |
135
|
|
|
|
|
|
|
|
136
|
|
|
|
|
|
|
=cut |
137
|
|
|
|
|
|
|
|