2 # BioPerl module for Bio::PrimarySeqI
4 # Please direct questions and support issues to <bioperl-l@bioperl.org>
6 # Cared for by Ewan Birney <birney@ebi.ac.uk>
8 # Copyright Ewan Birney
10 # You may distribute this module under the same terms as perl itself
12 # POD documentation - main docs before the code
17 Bio::PrimarySeqI - Interface definition for a Bio::PrimarySeq
21 # Bio::PrimarySeqI is the interface class for sequences.
22 # If you are a newcomer to bioperl, you might want to start with
23 # Bio::Seq documentation.
25 # Test if this is a seq object
26 $obj->isa("Bio::PrimarySeqI") ||
27 $obj->throw("$obj does not implement the Bio::PrimarySeqI interface");
30 $string = $obj->seq();
31 $substring = $obj->subseq(12,50);
32 $display = $obj->display_id(); # for human display
33 $id = $obj->primary_id(); # unique id for this object,
34 # implementation defined
35 $unique_key= $obj->accession_number(); # unique biological id
40 $rev = $obj->revcom();
43 $obj->throw( "Could not reverse complement. ".
44 "Probably not DNA. Actual exception\n$@\n" );
47 $trunc = $obj->trunc(12,50);
48 # $rev and $trunc are Bio::PrimarySeqI compliant objects
53 This object defines an abstract interface to basic sequence
54 information - for most users of the package the documentation (and
55 methods) in this class are not useful - this is a developers-only
56 class which defines what methods have to be implmented by other Perl
57 objects to comply to the Bio::PrimarySeqI interface. Go "perldoc
58 Bio::Seq" or "man Bio::Seq" for more information on the main class for
61 PrimarySeq is an object just for the sequence and its name(s), nothing
62 more. Seq is the larger object complete with features. There is a pure
63 perl implementation of this in L<Bio::PrimarySeq>. If you just want to
64 use L<Bio::PrimarySeq> objects, then please read that module first. This
65 module defines the interface, and is of more interest to people who
66 want to wrap their own Perl Objects/RDBs/FileSystems etc in way that
67 they "are" bioperl sequence objects, even though it is not using Perl
68 to store the sequence etc.
70 This interface defines what bioperl considers necessary to "be" a
71 sequence, without providing an implementation of this, an
72 implementation is provided in L<Bio::PrimarySeq>. If you want to provide
73 a Bio::PrimarySeq-compliant object which in fact wraps another
74 object/database/out-of-perl experience, then this is the correct thing
75 to wrap, generally by providing a wrapper class which would inherit
76 from your object and this Bio::PrimarySeqI interface. The wrapper class
77 then would have methods lists in the "Implementation Specific
78 Functions" which would provide these methods for your object.
84 User feedback is an integral part of the evolution of this and other
85 Bioperl modules. Send your comments and suggestions preferably to one
86 of the Bioperl mailing lists. Your participation is much appreciated.
88 bioperl-l@bioperl.org - General discussion
89 http://bioperl.org/wiki/Mailing_lists - About the mailing lists
93 Please direct usage questions or support issues to the mailing list:
95 I<bioperl-l@bioperl.org>
97 rather than to the module maintainer directly. Many experienced and
98 reponsive experts will be able look at the problem and quickly
99 address it. Please include a thorough description of the problem
100 with code and data examples if at all possible.
102 =head2 Reporting Bugs
104 Report bugs to the Bioperl bug tracking system to help us keep track
105 the bugs and their resolution. Bug reports can be submitted via the
108 https://redmine.open-bio.org/projects/bioperl/
110 =head1 AUTHOR - Ewan Birney
112 Email birney@ebi.ac.uk
116 The rest of the documentation details each of the object
117 methods. Internal methods are usually preceded with a _
122 package Bio
::PrimarySeqI
;
124 use Bio
::Tools
::CodonTable
;
126 use base
qw(Bio::Root::RootI);
129 =head1 Implementation-specific Functions
131 These functions are the ones that a specific implementation must
137 Usage : $string = $obj->seq()
138 Function: Returns the sequence as a string of letters. The
139 case of the letters is left up to the implementer.
140 Suggested cases are upper case for proteins and lower case for
141 DNA sequence (IUPAC standard), but implementations are suggested to
142 keep an open mind about case (some users... want mixed case!)
150 $self->throw_not_implemented();
157 Usage : $substring = $obj->subseq(10,40);
158 Function: Returns the subseq from start to end, where the first base
159 is 1 and the number is inclusive, i.e. 1-2 are the first two
160 bases of the sequence.
162 Start cannot be larger than end but can be equal.
172 $self->throw_not_implemented();
179 Usage : $id_string = $obj->display_id();
180 Function: Returns the display id, also known as the common name of the Sequence
183 The semantics of this is that it is the most likely string
184 to be used as an identifier of the sequence, and likely to
185 have "human" readability. The id is equivalent to the ID
186 field of the GenBank/EMBL databanks and the id field of the
187 Swissprot/sptrembl database. In fasta format, the >(\S+) is
188 presumed to be the id, though some people overload the id
189 to embed other information. Bioperl does not use any
190 embedded information in the ID field, and people are
191 encouraged to use other mechanisms (accession field for
192 example, or extending the sequence object) to solve this.
194 Notice that $seq->id() maps to this function, mainly for
195 legacy/convenience reasons.
204 $self->throw_not_implemented();
208 =head2 accession_number
210 Title : accession_number
211 Usage : $unique_biological_key = $obj->accession_number;
212 Function: Returns the unique biological id for a sequence, commonly
213 called the accession_number. For sequences from established
214 databases, the implementors should try to use the correct
215 accession number. Notice that primary_id() provides the
216 unique id for the implemetation, allowing multiple objects
217 to have the same accession number in a particular implementation.
219 For sequences with no accession number, this method should return
227 sub accession_number
{
228 my ($self,@args) = @_;
229 $self->throw_not_implemented();
236 Usage : $unique_implementation_key = $obj->primary_id;
237 Function: Returns the unique id for this object in this
238 implementation. This allows implementations to manage their
239 own object ids in a way the implementaiton can control
240 clients can expect one id to map to one object.
242 For sequences with no accession number, this method should
243 return a stringified memory location.
252 my ($self,@args) = @_;
253 $self->throw_not_implemented();
260 Usage : if( $obj->can_call_new ) {
261 $newobj = $obj->new( %param );
263 Function: Can_call_new returns 1 or 0 depending
264 on whether an implementation allows new
265 constructor to be called. If a new constructor
266 is allowed, then it should take the followed hashed
269 $myobject->new( -seq => $sequence_as_string,
271 -accession_number => $accession
281 my ($self,@args) = @_;
282 # we default to 0 here
290 Usage : if( $obj->alphabet eq 'dna' ) { /Do Something/ }
291 Function: Returns the type of sequence being one of
292 'dna', 'rna' or 'protein'. This is case sensitive.
294 This is not called "type" because this would cause
295 upgrade problems from the 0.5 and earlier Seq objects.
297 Returns : A string either 'dna','rna','protein'. NB - the object must
298 make a call of the alphabet, if there is no alphabet specified it
307 $self->throw_not_implemented();
314 Usage : Deprecated. Use alphabet() instead.
319 my ($self,@args) = @_;
320 $self->warn("moltype: pre v1.0 method. Calling alphabet() instead...");
321 return $self->alphabet(@args);
325 =head1 Implementation-optional Functions
327 The following functions rely on the above functions. An
328 implementing class does not need to provide these functions, as they
329 will be provided by this class, but is free to override these
332 The revcom(), trunc(), and translate() methods create new sequence
333 objects. They will call new() on the class of the sequence object
334 instance passed as argument, unless can_call_new() returns FALSE. In
335 the latter case a Bio::PrimarySeq object will be created. Implementors
336 which really want to control how objects are created (eg, for object
337 persistence over a database, or objects in a CORBA framework), they
338 are encouraged to override these methods
343 Usage : $rev = $seq->revcom()
344 Function: Produces a new Bio::PrimarySeqI implementing object which
345 is the reversed complement of the sequence. For protein
346 sequences this throws an exception of "Sequence is a
347 protein. Cannot revcom".
349 The id is the same id as the original sequence, and the
350 accession number is also indentical. If someone wants to
351 track that this sequence has be reversed, it needs to
352 define its own extensionsj.
354 To do an inplace edit of an object you can go:
356 $seq = $seq->revcom();
358 This of course, causes Perl to handle the garbage
359 collection of the old object, but it is roughly speaking as
360 efficient as an inplace edit.
362 Returns : A new (fresh) Bio::PrimarySeqI object
370 my ($seqclass, $opts) = $self->_setup_class;
371 my $out = $seqclass->new(
372 -seq
=> $self->_revcom_from_string($self->seq, $self->alphabet),
373 -is_circular
=> $self->is_circular,
374 -display_id
=> $self->display_id,
375 -accession_number
=> $self->accession_number,
376 -alphabet
=> $self->alphabet,
377 -desc
=> $self->desc,
378 -verbose
=> $self->verbose,
385 sub _revcom_from_string
{
386 my ($self, $string, $alphabet) = @_;
388 # Check that reverse-complementing makes sense
389 if( $alphabet eq 'protein' ) {
390 $self->throw("Sequence is a protein. Cannot revcom.");
392 if( $alphabet ne 'dna' && $alphabet ne 'rna' ) {
393 my $msg = "Sequence is not dna or rna, but [$alphabet]. Attempting to revcom, ".
394 "but unsure if this is right.";
395 if( $self->can('warn') ) {
398 warn("[$self] $msg");
402 # If sequence is RNA, map to DNA (then map back later)
403 if( $alphabet eq 'rna' ) {
404 $string =~ tr/uU/tT/;
407 # Reverse-complement now
408 $string =~ tr/acgtrymkswhbvdnxACGTRYMKSWHBVDNX/tgcayrkmswdvbhnxTGCAYRKMSWDVBHNX/;
409 $string = CORE
::reverse $string;
411 # Map back RNA to DNA
412 if( $alphabet eq 'rna' ) {
413 $string =~ tr/tT/uU/;
423 Usage : $subseq = $myseq->trunc(10,100);
424 Function: Provides a truncation of a sequence.
425 Returns : A fresh Bio::PrimarySeqI implementing object.
426 Args : Two integers denoting first and last base of the sub-sequence.
432 my ($self,$start,$end) = @_;
435 if( defined $start && ref($start) &&
436 $start->isa('Bio::LocationI') ) {
437 $str = $self->subseq($start); # start is a location actually
439 $self->throw("trunc start,end -- there was no end for $start");
440 } elsif( $end < $start ) {
441 my $msg = "start [$start] is greater than end [$end]. \n".
442 "If you want to truncated and reverse complement, \n".
443 "you must call trunc followed by revcom. Sorry.";
446 $str = $self->subseq($start,$end);
449 my ($seqclass, $opts) = $self->_setup_class;
450 my $out = $seqclass->new(
452 -display_id
=> $self->display_id,
453 -accession_number
=> $self->accession_number,
454 -alphabet
=> $self->alphabet,
455 -desc
=> $self->desc,
456 -verbose
=> $self->verbose,
466 Usage : $protein_seq_obj = $dna_seq_obj->translate
468 Or if you expect a complete coding sequence (CDS) translation,
469 with initiator at the beginning and terminator at the end:
471 $protein_seq_obj = $cds_seq_obj->translate(-complete => 1);
473 Or if you want translate() to find the first initiation
474 codon and return the corresponding protein:
476 $protein_seq_obj = $cds_seq_obj->translate(-orf => 1);
478 Function: Provides the translation of the DNA sequence using full
479 IUPAC ambiguities in DNA/RNA and amino acid codes.
481 The complete CDS translation is identical to EMBL/TREMBL
482 database translation. Note that the trailing terminator
483 character is removed before returning the translated protein
486 Note: if you set $dna_seq_obj->verbose(1) you will get a
487 warning if the first codon is not a valid initiator.
489 Returns : A Bio::PrimarySeqI implementing object
491 character for terminator, default '*'
493 character for unknown, default 'X'
495 positive integer frame shift (in bases), default 0
497 integer codon table id, default 1
499 boolean, if true, complete CDS is expected. default false
501 boolean, if true, codons which are incomplete are translated if a
502 suitable amino acid is found. For instance, if the incomplete
503 codon is 'GG', the completed codon is 'GGN', which is glycine
504 (G). Defaults to 'false'; setting '-complete' also makes this
507 boolean, throw exception if ORF not complete, default false
509 if 'longest', find longest ORF. other true value, find
512 optional L<Bio::Tools::CodonTable> object to use for
515 optional three-character string to force as initiation
516 codon (e.g. 'atg'). If unset, start codons are
517 determined by the CodonTable. Case insensitive.
519 optional positive integer offset for fuzzy locations.
520 if set, must be either 1, 2, or 3
524 The -start argument only applies when -orf is set to 1. By default all
525 initiation codons found in the given codon table are used but when
526 "start" is set to some codon this codon will be used exclusively as
527 the initiation codon. Note that the default codon table (NCBI
528 "Standard") has 3 initiation codons!
530 By default translate() translates termination codons to the some
531 character (default is *), both internal and trailing codons. Setting
532 "-complete" to 1 tells translate() to remove the trailing character.
534 -offset is used for seqfeatures which contain the the \codon_start tag
535 and can be set to 1, 2, or 3. This is the offset by which the
536 sequence translation starts relative to the first base of the feature
538 For details on codon tables used by translate() see L<Bio::Tools::CodonTable>.
540 Deprecated argument set (v. 1.5.1 and prior versions) where each argument is an
543 1: character for terminator (optional), defaults to '*'.
544 2: character for unknown amino acid (optional), defaults to 'X'.
545 3: frame (optional), valid values are 0, 1, 2, defaults to 0.
546 4: codon table id (optional), defaults to 1.
547 5: complete coding sequence expected, defaults to 0 (false).
548 6: boolean, throw exception if not complete coding sequence
549 (true), defaults to warning (false)
550 7: codontable, a custom Bio::Tools::CodonTable object (optional).
555 my ($self,@args) = @_;
556 my ($terminator, $unknown, $frame, $codonTableId, $complete,
557 $complete_codons, $throw, $codonTable, $orf, $start_codon, $offset);
559 ## new API with named parameters, post 1.5.1
560 if ($args[0] && $args[0] =~ /^-[A-Z]+/i) {
561 ($terminator, $unknown, $frame, $codonTableId, $complete,
562 $complete_codons, $throw,$codonTable, $orf, $start_codon, $offset) =
563 $self->_rearrange([qw(TERMINATOR
574 ## old API, 1.5.1 and preceding versions
576 ($terminator, $unknown, $frame, $codonTableId,
577 $complete, $throw, $codonTable, $offset) = @args;
580 ## Initialize termination codon, unknown codon, codon table id, frame
581 $terminator = '*' unless (defined($terminator) and $terminator ne '');
582 $unknown = "X" unless (defined($unknown) and $unknown ne '');
583 $frame = 0 unless (defined($frame) and $frame ne '');
584 $codonTableId = 1 unless (defined($codonTableId) and $codonTableId ne '');
585 $complete_codons ||= $complete || 0;
587 ## Get a CodonTable, error if custom CodonTable is invalid
589 $self->throw("Need a Bio::Tools::CodonTable object, not ". $codonTable)
590 unless $codonTable->isa('Bio::Tools::CodonTable');
593 # shouldn't this be cached? Seems wasteful to have a new instance
595 $codonTable = Bio
::Tools
::CodonTable
->new( -id
=> $codonTableId);
598 ## Error if alphabet is "protein"
599 $self->throw("Can't translate an amino acid sequence.") if
600 ($self->alphabet =~ /protein/i);
602 ## Error if -start parameter isn't a valid codon
604 $self->throw("Invalid start codon: $start_codon.") if
605 ( $start_codon !~ /^[A-Z]{3}$/i );
610 $self->throw("Offset must be 1, 2, or 3.") if
611 ( $offset !~ /^[123]$/ );
612 my ($start, $end) = ($offset, $self->length);
613 ($seq) = $self->subseq($start, $end);
615 ($seq) = $self->seq();
618 ## ignore frame if an ORF is supposed to be found
620 my ($orf_region) = $self->_find_orfs_nucleotide( $seq, $codonTable, $start_codon, $orf eq 'longest' ?
0 : 'first_only' );
621 $seq = $self->_orf_sequence( $seq, $orf_region );
623 ## use frame, error if frame is not 0, 1 or 2
624 $self->throw("Valid values for frame are 0, 1, or 2, not $frame.")
625 unless ($frame == 0 or $frame == 1 or $frame == 2);
626 $seq = substr($seq,$frame);
630 my $output = $codonTable->translate($seq, $complete_codons);
631 # Use user-input terminator/unknown
632 $output =~ s/\*/$terminator/g;
633 $output =~ s/X/$unknown/g;
635 ## Only if we are expecting to translate a complete coding region
637 my $id = $self->display_id;
638 # remove the terminator character
639 if( substr($output,-1,1) eq $terminator ) {
642 $throw && $self->throw("Seq [$id]: Not using a valid terminator codon!");
643 $self->warn("Seq [$id]: Not using a valid terminator codon!");
645 # test if there are terminator characters inside the protein sequence!
646 if ($output =~ /\Q$terminator\E/) {
648 $throw && $self->throw("Seq [$id]: Terminator codon inside CDS!");
649 $self->warn("Seq [$id]: Terminator codon inside CDS!");
651 # if the initiator codon is not ATG, the amino acid needs to be changed to M
652 if ( substr($output,0,1) ne 'M' ) {
653 if ($codonTable->is_start_codon(substr($seq, 0, 3)) ) {
654 $output = 'M'. substr($output,1);
656 $self->throw("Seq [$id]: Not using a valid initiator codon!");
658 $self->warn("Seq [$id]: Not using a valid initiator codon!");
663 my ($seqclass, $opts) = $self->_setup_class;
664 my $out = $seqclass->new(
666 -display_id
=> $self->display_id,
667 -accession_number
=> $self->accession_number,
668 # is there anything wrong with retaining the desc?
669 -desc
=> $self->desc,
670 -alphabet
=> 'protein',
671 -verbose
=> $self->verbose,
681 Usage : $xseq = $seq->transcribe
682 Function: Convert base T to base U
683 Returns : PrimarySeqI object of alphabet 'rna' or
684 undef if $seq->alphabet ne 'dna'
691 return unless $self->alphabet eq 'dna';
694 my $desc = $self->desc || '';
695 my ($seqclass, $opts) = $self->_setup_class;
696 return $seqclass->new(
699 -display_id
=> $self->display_id,
700 -accession_number
=> $self->accession_number,
701 -desc
=> "${desc}[TRANSCRIBED]",
702 -verbose
=> $self->verbose,
708 =head2 rev_transcribe()
710 Title : rev_transcribe
711 Usage : $rtseq = $seq->rev_transcribe
712 Function: Convert base U to base T
713 Returns : PrimarySeqI object of alphabet 'dna' or
714 undef if $seq->alphabet ne 'rna'
721 return unless $self->alphabet eq 'rna';
724 my ($seqclass, $opts) = $self->_setup_class;
725 return $seqclass->new(
728 -display_id
=> $self->display_id,
729 -accession_number
=> $self->accession_number,
730 -desc
=> $self->desc . "[REVERSE TRANSCRIBED]",
731 -verbose
=> $self->verbose,
740 Usage : $id = $seq->id()
741 Function: ID of the sequence. This should normally be (and actually is in
742 the implementation provided here) just a synonym for display_id().
750 return $self->display_id();
757 Usage : $len = $seq->length()
759 Returns : Integer representing the length of the sequence.
766 $self->throw_not_implemented();
773 Usage : $seq->desc($newval);
774 $description = $seq->desc();
775 Function: Get/set description text for a seq object
776 Returns : Value of desc
777 Args : newvalue (optional)
782 shift->throw_not_implemented();
789 Usage : if( $obj->is_circular) { # Do something }
790 Function: Returns true if the molecule is circular
791 Returns : Boolean value
797 shift->throw_not_implemented;
801 =head1 Private functions
803 These are some private functions for the PrimarySeqI interface. You do not
804 need to implement these functions
806 =head2 _find_orfs_nucleotide
808 Title : _find_orfs_nucleotide
810 Function: Finds ORF starting at 1st initiation codon in nucleotide sequence.
811 The ORF is not required to have a termination codon.
813 Returns : a list of string coordinates of ORF locations (0-based half-open),
814 sorted descending by length (so that the longest is first)
815 as: [ start, end, frame, length ], [ start, end, frame, length ], ...
816 Args : Nucleotide sequence,
818 (optional) alternative initiation codon (e.g. 'ATA'),
819 (optional) boolean that, if true, stops after finding the
824 sub _find_orfs_nucleotide
{
825 my ( $self, $sequence, $codon_table, $start_codon, $first_only ) = @_;
826 $sequence = uc $sequence;
827 $start_codon = uc $start_codon if $start_codon;
829 my $is_start = $start_codon
830 ?
sub { shift eq $start_codon }
831 : sub { $codon_table->is_start_codon( shift ) };
833 # stores the begin index of the currently-running ORF in each
835 my @current_orf_start = (-1,-1,-1);
837 #< stores coordinates of longest observed orf (so far) in each
841 # go through each base of the sequence, and each reading frame for each base
842 my $seqlen = CORE
::length $sequence;
843 for( my $j = 0; $j <= $seqlen-3; $j++ ) {
846 my $this_codon = substr( $sequence, $j, 3 );
848 # if in an orf and this is either a stop codon or the last in-frame codon in the string
849 if ( $current_orf_start[$frame] >= 0 ) {
850 if ( $codon_table->is_ter_codon( $this_codon ) ||( my $is_last_codon_in_frame = ($j >= $seqlen-5)) ) {
851 # record ORF start, end (half-open), length, and frame
852 my @this_orf = ( $current_orf_start[$frame], $j+3, undef, $frame );
853 my $this_orf_length = $this_orf[2] = ( $this_orf[1] - $this_orf[0] );
855 $self->warn( "Translating partial ORF "
856 .$self->_truncate_seq( $self->_orf_sequence( $sequence, \
@this_orf ))
857 .' from end of nucleotide sequence'
859 if $first_only && $is_last_codon_in_frame;
861 return \
@this_orf if $first_only;
862 push @orfs, \
@this_orf;
863 $current_orf_start[$frame] = -1;
866 # if this is a start codon
867 elsif ( $is_start->($this_codon) ) {
868 $current_orf_start[$frame] = $j;
872 return sort { $b->[2] <=> $a->[2] } @orfs;
877 my ($self, $seq) = @_;
878 return CORE
::length($seq) > 200 ?
substr($seq,0,50).'...'.substr($seq,-50) : $seq;
883 my ($self, $seq, $orf ) = @_;
884 return '' unless $orf;
885 return substr( $seq, $orf->[0], $orf->[2] )
889 =head2 _attempt_to_load_Seq
891 Title : _attempt_to_load_Seq
900 sub _attempt_to_load_Seq
{
903 if( $main::{'Bio::PrimarySeq'} ) {
907 require Bio
::PrimarySeq
;
910 my $text = "Bio::PrimarySeq could not be loaded for [$self]\n".
911 "This indicates that you are using Bio::PrimarySeqI ".
912 "without Bio::PrimarySeq loaded or without providing a ".
913 "complete implementation.\nThe most likely problem is that there ".
914 "has been a misconfiguration of the bioperl environment\n".
915 "Actual exception:\n\n";
916 $self->throw("$text$@\n");
925 # Return name of class and setup some default parameters
928 if ($self->can_call_new()) {
929 $seqclass = ref($self);
931 $seqclass = 'Bio::PrimarySeq';
932 $self->_attempt_to_load_Seq();
935 if ($seqclass eq 'Bio::PrimarySeq') {
936 # Since sequence is in a Seq object, it has already been validated.
937 # We do not need to validate its trunc(), revcom(), etc
938 $opts{ -direct
} = 1;
940 return $seqclass, \
%opts;