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# POD documentation - main docs before the code |
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3
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=head1 NAME |
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GenOO::Spliceable - Role for a region that can be spliced |
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7
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=head1 SYNOPSIS |
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# This role provides regions with the splicing attributes and methods |
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=head1 DESCRIPTION |
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13
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An object that consumes this role gets splicing attributes and methods such as exons and |
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introns. The key attributes of this class are "splice_starts" and "splice_stops" |
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which are sorted arrays of coordinates that define the intervals for exons. |
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-------------EXON_1----------- ------------EXON_2------------ |
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SPLICE_START_1...SPLICE_STOP_1...INTRON...SPLICE_START_2...SPLICE_STOP_2...INTRON... |
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=head1 EXAMPLES |
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# Get the location information on the reference sequence |
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$obj_with_role->exons; |
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$obj_with_role->introns; |
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26
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# Check if a position is within an exon or an intron |
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$obj_with_role->is_position_within_exon(120); # 1/0 |
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$obj_with_role->is_position_within_intron(120); # 0/1 |
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29
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30
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# Get the length of the exonic region |
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31
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$obj_with_role->exonic_length; |
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32
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33
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=cut |
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34
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35
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# Let the code begin... |
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36
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37
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package GenOO::Spliceable; |
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38
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$GenOO::Spliceable::VERSION = '1.4.6'; |
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39
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1
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1
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636
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use Moose::Role; |
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1
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1
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1
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8
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40
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1
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1
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3914
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use Moose::Util::TypeConstraints; |
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1
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1
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1
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7
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41
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1
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1
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1551
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use namespace::autoclean; |
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2
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1
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10
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42
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43
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1
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1
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410
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use GenOO::Exon; |
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1
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2
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1
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41
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44
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1
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1
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482
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use GenOO::Intron; |
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1
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3
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1
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40
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45
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1
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1
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495
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use GenOO::Junction; |
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1
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2
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1
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1310
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46
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47
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# Define new data type |
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48
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subtype 'SortedArrayRef', as 'ArrayRef', where { _sorted_array() }; |
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49
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50
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# Define coercions to new data type |
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51
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coerce 'SortedArrayRef', from 'ArrayRef', via { [sort {$a <=> $b} @{$_}] }; |
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52
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coerce 'SortedArrayRef', from 'Str' , via { [sort {$a <=> $b} (split(/\D+/,$_))] }; |
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53
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54
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# Define attributes |
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55
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has 'splice_starts' => ( |
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56
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isa => 'SortedArrayRef', |
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57
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is => 'ro', |
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58
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writer => '_set_splice_starts', |
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59
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required => 1, |
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60
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coerce => 1 |
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61
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); |
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62
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63
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has 'splice_stops' => ( |
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64
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isa => 'SortedArrayRef', |
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65
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is => 'ro', |
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66
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writer => '_set_splice_stops', |
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67
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required => 1, |
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68
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coerce => 1 |
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69
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); |
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70
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71
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has 'exons' => ( |
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72
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isa => 'ArrayRef', |
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73
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is => 'ro', |
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74
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builder => '_create_exons', |
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75
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init_arg => undef, |
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76
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lazy => 1, |
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77
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); |
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78
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79
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has 'introns' => ( |
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80
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isa => 'ArrayRef', |
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81
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is => 'ro', |
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82
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builder => '_create_introns', |
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83
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init_arg => undef, |
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84
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lazy => 1, |
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85
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); |
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86
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87
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# Define consumed roles |
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88
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with 'GenOO::Region'; |
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89
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90
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91
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sub BUILD { |
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92
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1049
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1049
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0
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1334
|
my $self = shift; |
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93
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94
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1049
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2882
|
$self->_sanitize_splice_starts_and_stops; |
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95
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} |
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96
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97
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####################################################################### |
|
98
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######################## Interface Methods ######################## |
|
99
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####################################################################### |
|
100
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sub is_position_within_exon { |
|
101
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4
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4
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0
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600
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my ($self, $position) = @_; |
|
102
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103
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4
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144
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my $exons = $self->exons; |
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104
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4
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12
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foreach my $exon (@$exons) { |
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105
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10
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100
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33
|
if ($exon->contains_position($position)) { |
|
106
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2
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8
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return 1; |
|
107
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} |
|
108
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} |
|
109
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2
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14
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return 0; |
|
110
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} |
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111
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112
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sub is_position_within_intron { |
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113
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2
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2
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0
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643
|
my ($self, $position) = @_; |
|
114
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115
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2
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66
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my $introns = $self->introns; |
|
116
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2
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8
|
foreach my $intron (@$introns) { |
|
117
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4
|
100
|
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16
|
if ($intron->contains_position($position)) { |
|
118
|
1
|
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5
|
return 1; |
|
119
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} |
|
120
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} |
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121
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1
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8
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return 0; |
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122
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} |
|
123
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124
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sub exon_exon_junctions { |
|
125
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1
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1
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0
|
646
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my ($self) = @_; |
|
126
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127
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1
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4
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my @junctions; |
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128
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my @junction_starts; |
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129
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0
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0
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my @junction_stops; |
|
130
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131
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1
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39
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my $exons = $self->exons; |
|
132
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1
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50
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7
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if (@$exons > 1) { |
|
133
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1
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8
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for (my $i=0;$i<@$exons-1;$i++) { |
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134
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3
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83
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push @junction_starts, $$exons[$i]->stop; |
|
135
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3
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77
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push @junction_stops, $$exons[$i+1]->start; |
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136
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} |
|
137
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} |
|
138
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139
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1
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50
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8
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my $junctions_count = @junction_starts == @junction_stops ? @junction_starts : die "Junctions starts are not of the same size as junction stops\n"; |
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140
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1
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8
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for (my $i=0;$i<$junctions_count;$i++) { |
|
141
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3
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77
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push @junctions, GenOO::Junction->new( |
|
142
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species => $self->species, |
|
143
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strand => $self->strand, |
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144
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chromosome => $self->chromosome, |
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145
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start => $junction_starts[$i], |
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146
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stop => $junction_stops[$i], |
|
147
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part_of => $self, |
|
148
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); |
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149
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} |
|
150
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1
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7
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return \@junctions; |
|
151
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} |
|
152
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153
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sub exonic_sequence { |
|
154
|
1
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1
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0
|
598
|
my ($self) = @_; |
|
155
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156
|
1
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50
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45
|
if (defined $self->sequence) { |
|
157
|
1
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3
|
my $exonic_sequence = ''; |
|
158
|
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159
|
1
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50
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35
|
my $seq = $self->strand == 1 ? $self->sequence : reverse($self->sequence); |
|
160
|
1
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2
|
foreach my $exon (@{$self->exons}) { |
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1
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27
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161
|
4
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96
|
$exonic_sequence .= substr($seq, ($exon->start - $self->start), $exon->length); |
|
162
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} |
|
163
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164
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1
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50
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28
|
if ($self->strand == 1) { |
|
165
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0
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0
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return $exonic_sequence; |
|
166
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} |
|
167
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else { |
|
168
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1
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|
8
|
return reverse($exonic_sequence); |
|
169
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} |
|
170
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} |
|
171
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} |
|
172
|
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173
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sub exonic_length { |
|
174
|
1
|
|
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1
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0
|
628
|
my ($self) = @_; |
|
175
|
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176
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1
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2
|
my $length = 0; |
|
177
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1
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3
|
foreach my $exon (@{$self->exons}) { |
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1
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34
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178
|
4
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|
97
|
$length += $exon->length; |
|
179
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} |
|
180
|
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181
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1
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|
7
|
return $length; |
|
182
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} |
|
183
|
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184
|
|
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sub intronic_length { |
|
185
|
1
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|
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1
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0
|
671
|
my ($self) = @_; |
|
186
|
|
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187
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1
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2
|
my $length = 0; |
|
188
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1
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1
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foreach my $intron (@{$self->introns}) { |
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1
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39
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189
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3
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73
|
$length += $intron->length; |
|
190
|
|
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} |
|
191
|
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192
|
1
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|
5
|
return $length; |
|
193
|
|
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} |
|
194
|
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195
|
|
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|
|
sub relative_exonic_position { |
|
196
|
2
|
|
|
2
|
0
|
665
|
my ($self, $abs_pos) = @_; |
|
197
|
|
|
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|
198
|
2
|
100
|
|
|
|
7
|
if ($self->is_position_within_exon($abs_pos)) { |
|
199
|
1
|
|
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|
|
26
|
my $relative_pos = $abs_pos - $self->start; |
|
200
|
1
|
|
|
|
|
1
|
foreach my $intron (@{$self->introns}) { |
|
|
1
|
|
|
|
|
30
|
|
|
201
|
1
|
50
|
|
|
|
24
|
if ($intron->stop < $abs_pos) { |
|
202
|
0
|
|
|
|
|
0
|
$relative_pos -= $intron->length; |
|
203
|
|
|
|
|
|
|
} |
|
204
|
|
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|
|
else { |
|
205
|
1
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|
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|
2
|
last; |
|
206
|
|
|
|
|
|
|
} |
|
207
|
|
|
|
|
|
|
} |
|
208
|
1
|
|
|
|
|
8
|
return $relative_pos; |
|
209
|
|
|
|
|
|
|
} |
|
210
|
|
|
|
|
|
|
else { |
|
211
|
1
|
|
|
|
|
5
|
return undef; |
|
212
|
|
|
|
|
|
|
} |
|
213
|
|
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|
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|
|
} |
|
214
|
|
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|
|
215
|
|
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|
|
|
|
sub set_splice_starts_and_stops { |
|
216
|
966
|
|
|
966
|
0
|
1111
|
my ($self, $splice_starts, $splice_stops) = @_; |
|
217
|
|
|
|
|
|
|
|
|
218
|
966
|
|
|
|
|
26606
|
$self->_set_splice_starts($splice_starts); |
|
219
|
966
|
|
|
|
|
25677
|
$self->_set_splice_stops($splice_stops); |
|
220
|
966
|
|
|
|
|
1898
|
$self->_sanitize_splice_starts_and_stops; |
|
221
|
|
|
|
|
|
|
} |
|
222
|
|
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|
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|
|
223
|
|
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|
|
|
####################################################################### |
|
224
|
|
|
|
|
|
|
####################### Private Methods ############################ |
|
225
|
|
|
|
|
|
|
####################################################################### |
|
226
|
|
|
|
|
|
|
sub _create_exons { |
|
227
|
16
|
|
|
16
|
|
26
|
my ($self) = @_; |
|
228
|
|
|
|
|
|
|
|
|
229
|
16
|
|
|
|
|
392
|
my $exon_starts = $self->splice_starts; |
|
230
|
16
|
|
|
|
|
384
|
my $exon_stops = $self->splice_stops; |
|
231
|
|
|
|
|
|
|
|
|
232
|
16
|
|
|
|
|
17
|
my @exons; |
|
233
|
16
|
|
|
|
|
29
|
for (my $i=0;$i<@{$exon_starts};$i++) { |
|
|
62
|
|
|
|
|
140
|
|
|
234
|
46
|
|
|
|
|
1168
|
push @exons, GenOO::Exon->new({ |
|
235
|
|
|
|
|
|
|
strand => $self->strand, |
|
236
|
|
|
|
|
|
|
chromosome => $self->rname, |
|
237
|
|
|
|
|
|
|
start => $$exon_starts[$i], |
|
238
|
|
|
|
|
|
|
stop => $$exon_stops[$i], |
|
239
|
|
|
|
|
|
|
part_of => $self |
|
240
|
|
|
|
|
|
|
}); |
|
241
|
|
|
|
|
|
|
} |
|
242
|
|
|
|
|
|
|
|
|
243
|
16
|
|
|
|
|
375
|
return \@exons; |
|
244
|
|
|
|
|
|
|
} |
|
245
|
|
|
|
|
|
|
|
|
246
|
|
|
|
|
|
|
sub _create_introns { |
|
247
|
3
|
|
|
3
|
|
5
|
my ($self) = @_; |
|
248
|
|
|
|
|
|
|
|
|
249
|
3
|
|
|
|
|
75
|
my $exon_starts = $self->splice_starts; |
|
250
|
3
|
|
|
|
|
72
|
my $exon_stops = $self->splice_stops; |
|
251
|
|
|
|
|
|
|
|
|
252
|
3
|
|
|
|
|
3
|
my @introns; |
|
253
|
|
|
|
|
|
|
|
|
254
|
3
|
50
|
|
|
|
76
|
if ($self->start < $$exon_starts[0]) { |
|
255
|
0
|
|
|
|
|
0
|
push @introns, GenOO::Intron->new({ |
|
256
|
|
|
|
|
|
|
strand => $self->strand, |
|
257
|
|
|
|
|
|
|
chromosome => $self->rname, |
|
258
|
|
|
|
|
|
|
start => $self->start, |
|
259
|
|
|
|
|
|
|
stop => $$exon_starts[0] - 1, |
|
260
|
|
|
|
|
|
|
part_of => $self, |
|
261
|
|
|
|
|
|
|
}); |
|
262
|
|
|
|
|
|
|
} |
|
263
|
|
|
|
|
|
|
|
|
264
|
3
|
|
|
|
|
5
|
for (my $i=1;$i<@{$exon_starts};$i++) { |
|
|
12
|
|
|
|
|
24
|
|
|
265
|
9
|
|
|
|
|
17
|
push @introns, (GenOO::Intron->new({ |
|
266
|
|
|
|
|
|
|
strand => $self->strand, |
|
267
|
|
|
|
|
|
|
chromosome => $self->rname, |
|
268
|
9
|
|
|
|
|
240
|
start => ${$exon_stops}[$i-1] + 1, |
|
269
|
9
|
|
|
|
|
245
|
stop => ${$exon_starts}[$i] - 1, |
|
270
|
|
|
|
|
|
|
part_of => $self, |
|
271
|
|
|
|
|
|
|
})); |
|
272
|
|
|
|
|
|
|
} |
|
273
|
|
|
|
|
|
|
|
|
274
|
3
|
50
|
|
|
|
78
|
if ($self->stop > $$exon_stops[-1]) { |
|
275
|
0
|
|
|
|
|
0
|
push @introns, (GenOO::Intron->new({ |
|
276
|
|
|
|
|
|
|
strand => $self->strand, |
|
277
|
|
|
|
|
|
|
chromosome => $self->rname, |
|
278
|
|
|
|
|
|
|
start => $$exon_stops[-1] + 1, |
|
279
|
|
|
|
|
|
|
stop => $self->stop, |
|
280
|
|
|
|
|
|
|
part_of => $self, |
|
281
|
|
|
|
|
|
|
})); |
|
282
|
|
|
|
|
|
|
} |
|
283
|
|
|
|
|
|
|
|
|
284
|
3
|
|
|
|
|
76
|
return \@introns; |
|
285
|
|
|
|
|
|
|
} |
|
286
|
|
|
|
|
|
|
|
|
287
|
|
|
|
|
|
|
sub _sanitize_splice_starts_and_stops { |
|
288
|
2015
|
|
|
2015
|
|
2063
|
my ($self) = @_; |
|
289
|
|
|
|
|
|
|
|
|
290
|
2015
|
|
|
|
|
46813
|
my $splice_starts = $self->splice_starts; |
|
291
|
2015
|
|
|
|
|
45560
|
my $splice_stops = $self->splice_stops; |
|
292
|
|
|
|
|
|
|
|
|
293
|
2015
|
50
|
|
|
|
4275
|
if (@$splice_starts != @$splice_stops) { |
|
294
|
0
|
|
|
|
|
0
|
die "Error: Spice starts array is not of the same size as splice_stops (".scalar @$splice_starts."!=".scalar @$splice_stops.")\n"; |
|
295
|
|
|
|
|
|
|
} |
|
296
|
|
|
|
|
|
|
|
|
297
|
2015
|
|
|
|
|
2411
|
my $index = 0; |
|
298
|
2015
|
|
|
|
|
25655
|
while ($index < (@$splice_starts-1)) { |
|
299
|
5547
|
50
|
|
|
|
6989
|
if ($$splice_stops[$index] == $$splice_starts[$index+1] - 1) { |
|
300
|
0
|
|
|
|
|
0
|
$$splice_stops[$index] = $$splice_stops[$index+1]; |
|
301
|
0
|
|
|
|
|
0
|
splice @$splice_starts, $index+1, 1; |
|
302
|
0
|
|
|
|
|
0
|
splice @$splice_stops, $index+1, 1; |
|
303
|
|
|
|
|
|
|
} |
|
304
|
|
|
|
|
|
|
else { |
|
305
|
5547
|
|
|
|
|
9971
|
$index++; |
|
306
|
|
|
|
|
|
|
} |
|
307
|
|
|
|
|
|
|
} |
|
308
|
|
|
|
|
|
|
} |
|
309
|
|
|
|
|
|
|
|
|
310
|
|
|
|
|
|
|
####################################################################### |
|
311
|
|
|
|
|
|
|
####################### Private Routines ########################### |
|
312
|
|
|
|
|
|
|
####################################################################### |
|
313
|
|
|
|
|
|
|
sub _sanitize_splice_coords_within_limits { |
|
314
|
6
|
|
|
6
|
|
7
|
my ($pre_splice_starts, $pre_splice_stops, $start, $stop) = @_; |
|
315
|
|
|
|
|
|
|
|
|
316
|
6
|
|
|
|
|
7
|
my @splice_starts; |
|
317
|
|
|
|
|
|
|
my @splice_stops; |
|
318
|
6
|
|
|
|
|
15
|
for (my $i=0;$i<@$pre_splice_starts;$i++) { |
|
319
|
48
|
100
|
|
|
|
74
|
if ($$pre_splice_stops[$i] < $start) { |
|
|
|
100
|
|
|
|
|
|
|
320
|
14
|
|
|
|
|
20
|
next; |
|
321
|
|
|
|
|
|
|
} |
|
322
|
|
|
|
|
|
|
elsif ($$pre_splice_starts[$i] > $stop) { |
|
323
|
14
|
|
|
|
|
19
|
next; |
|
324
|
|
|
|
|
|
|
} |
|
325
|
|
|
|
|
|
|
else { #if the exon overlaps or is contained in the UTR5 |
|
326
|
20
|
100
|
|
|
|
27
|
if ($start >= $$pre_splice_starts[$i]) { |
|
327
|
6
|
|
|
|
|
9
|
push @splice_starts, $start; |
|
328
|
|
|
|
|
|
|
} |
|
329
|
|
|
|
|
|
|
else { |
|
330
|
14
|
|
|
|
|
12
|
push @splice_starts, $$pre_splice_starts[$i]; |
|
331
|
|
|
|
|
|
|
} |
|
332
|
20
|
100
|
|
|
|
25
|
if ($stop < $$pre_splice_stops[$i]) { |
|
333
|
4
|
|
|
|
|
8
|
push @splice_stops, $stop; |
|
334
|
|
|
|
|
|
|
} |
|
335
|
|
|
|
|
|
|
else { |
|
336
|
16
|
|
|
|
|
25
|
push @splice_stops, $$pre_splice_stops[$i]; |
|
337
|
|
|
|
|
|
|
} |
|
338
|
|
|
|
|
|
|
} |
|
339
|
|
|
|
|
|
|
} |
|
340
|
6
|
|
|
|
|
19
|
return \@splice_starts, \@splice_stops; |
|
341
|
|
|
|
|
|
|
} |
|
342
|
|
|
|
|
|
|
|
|
343
|
|
|
|
|
|
|
sub _sorted_array { |
|
344
|
8060
|
|
|
8060
|
|
6942
|
my $arrayref = $_; |
|
345
|
|
|
|
|
|
|
|
|
346
|
8060
|
100
|
|
|
|
5897
|
if (@{$arrayref} > 1){ |
|
|
8060
|
|
|
|
|
13801
|
|
|
347
|
3472
|
|
|
|
|
2965
|
for (my $i = 1; $i < @{$arrayref}; $i++){ |
|
|
25620
|
|
|
|
|
33315
|
|
|
348
|
22168
|
100
|
|
|
|
34740
|
if ($$arrayref[$i] < $$arrayref[$i-1]){ |
|
349
|
20
|
|
|
|
|
51
|
return 0; |
|
350
|
|
|
|
|
|
|
} |
|
351
|
|
|
|
|
|
|
} |
|
352
|
3452
|
|
|
|
|
6082
|
return 1; |
|
353
|
|
|
|
|
|
|
} |
|
354
|
4588
|
|
|
|
|
7310
|
return 1; |
|
355
|
|
|
|
|
|
|
} |
|
356
|
|
|
|
|
|
|
|
|
357
|
|
|
|
|
|
|
1; |