#
# BioPerl module for Bio::Restriction::IO::base
#
# Please direct questions and support issues to <bioperl-l@bioperl.org>
#
# Cared for by Rob Edwards <redwards@utmem.edu>
#
# Copyright Rob Edwards
#
# You may distribute this module under the same terms as perl itself
#
# POD documentation - main docs before the code
=head1 NAME
Bio::Restriction::IO::base - base enzyme set
=head1 SYNOPSIS
Do not use this module directly. Use it via the Bio::Restriction::IO class.
=head1 DESCRIPTION
This class defines some base methods for restriction enzyme input and
at the same time gives a base list of common enzymes.
=head1 FEEDBACK
=head2 Mailing Lists
User feedback is an integral part of the evolution of this and other
Bioperl modules. Send your comments and suggestions preferably to the
Bioperl mailing lists Your participation is much appreciated.
bioperl-l@bioperl.org - General discussion
http://bioperl.org/wiki/Mailing_lists - About the mailing lists
=head2 Support
Please direct usage questions or support issues to the mailing list:
I<bioperl-l@bioperl.org>
rather than to the module maintainer directly. Many experienced and
reponsive experts will be able look at the problem and quickly
address it. Please include a thorough description of the problem
with code and data examples if at all possible.
=head2 Reporting Bugs
Report bugs to the Bioperl bug tracking system to help us keep track
the bugs and their resolution. Bug reports can be submitted via the
web:
https://redmine.open-bio.org/projects/bioperl/
=head1 AUTHOR
Rob Edwards, redwards@utmem.edu
=head1 CONTRIBUTORS
Heikki Lehvaslaiho, heikki-at-bioperl-dot-org
Mark A. Jensen, maj-at-fortinbras-dot-us
=head1 APPENDIX
The rest of the documentation details each of the object
methods. Internal methods are usually preceded with a _
=cut
# Let the code begin...
package Bio::Restriction::IO::base;
use strict;
use Bio::Restriction::Enzyme;
use Bio::Restriction::EnzymeCollection;
use Bio::Restriction::Enzyme::MultiCut;
use Bio::Restriction::Enzyme::MultiSite;
use base qw(Bio::Restriction::IO);
my $offset; # class variable
sub new {
my($class, @args) = @_;
$class = ref $class ? ref $class : $class;
my $self = bless {}, $class;
$self->_initialize(@args);
return $self;
}
{
my %FILE_FORMAT = (
#'itype2' => 'itype2', # itype2 format doesn't work with 'current'
#'8' => 'itype2',
'withrefm' => 'withrefm',
'31' => 'withrefm',
#'bairoch' => 'bairoch', # bairoch format doesn't work with 'current'
#'19' => 'bairoch',
#'macvector' => 'bairoch',
#'vectorNTI' => 'bairoch',
'neo' => 'neos',
'prototype' => 'proto'
);
sub _initialize {
my($self,@args) = @_;
my ($current, $url, $file, $fh, $format, $verbose) =
$self->_rearrange([qw(CURRENT URL FILE FH FORMAT VERBOSE)],@args);
$verbose || 0;
$self->verbose($verbose);
if ($current && $format) {
$self->throw("Can't use -current with file, fh, or url set") if ($url || $file || $fh);
$self->throw("Format $format not retrievable using 'current'") if (!exists $FILE_FORMAT{$format});
my $io = $self->new(-url => 'ftp://ftp.neb.com/pub/rebase/VERSION');
chomp (my $version = $io->_readline);
push @args, (-url => "ftp://ftp.neb.com/pub/rebase/$FILE_FORMAT{$format}.$version", -retries => 1);
}
$self->_companies;
return unless $self->SUPER::_initialize(@args);
}
}
=head2 read
Title : read
Usage : $renzs = $stream->read
Function: reads all the restrction enzymes from the stream
Returns : a Bio::Restriction::Restriction object
Args : none
=cut
sub read {
my $self = shift;
my $renzs = Bio::Restriction::EnzymeCollection->new(-empty => 1);
seek DATA,($offset||=tell DATA), 0;
while (<DATA>) {
chomp;
next if /^\s*$/;
my ($name, $site, $cut) = split /\s+/;
my $re = Bio::Restriction::Enzyme->new(-name => $name,
-site => $site,
-cut => $cut);
$renzs->enzymes($re);
}
return $renzs;
}
=head2 _xln_sub
Title : _xln_sub
Function: Translates withrefm coords to Bio::Restriction coords
Args : Bio::Restriction::Enzyme object, scalar integer (cut posn)
Note : Used internally; pass as a coderef to the B:R::Enzyme
constructor
Note : It is convenient for each format module to have its own
version of this; not currently demanded by the interface.
=cut
sub _xln_sub { # for base.pm, a no-op
my ($z,$c) = @_;
return $c;
}
=head2 write
Title : write
Usage : $stream->write($renzs)
Function: writes restriction enzymes into the stream
Returns : 1 for success and 0 for error
Args : a Bio::Restriction::Enzyme
or a Bio::Restriction::EnzymeCollection object
=cut
sub write {
my $self = shift;
foreach (@_) {
map { printf "%s\t%s\t%s\n", $_->name, $_->string, $_->cut
} sort {$a->name cmp $b->name} $_->each_enzyme
if $_->isa('Bio::Restriction::EnzymeCollection');
printf "%s\t%s\t%s\n", $_->name, $_->string, $_->cut
if $_->isa('Bio::Restriction::Enzyme');
}
}
=head2 verify_prototype
Title : verify_prototype
Purpose : checks enzyme against current prototype list (retrieved remotely)
Returns : returns TRUE if enzyme is prototype
Argument : Bio::Restriction::EnzymeI
Comments : This is an auxiliary method to retrieve and check an enzyme
as a prototype. It retrieves the current list, stores it
as a singleton instance, then uses it to check the prototype
and modify is_prototype() to true or false. Use as follows:
my $col = $io->read;
for my $enz ($col->each_enzyme) {
print $enz->name.":".$enz->site."\n";
print "\t".$io->verify_prototype($enz)."\n";
}
=cut
my $protodb;
sub verify_prototype {
my ($self, $enz) = @_;
$self->throw("Must pass a Bio::Restriction::EnzymeI") unless
$enz && ref $enz && $enz->isa("Bio::Restriction::EnzymeI");
if (!defined $protodb) {
my $io = Bio::Restriction::IO->new(-format => 'prototype',
-current => 1);
$protodb = $io->read;
}
if ($protodb->get_enzyme($enz->name)) {
$enz->is_prototype(1);
} else {
$enz->is_prototype(0);
}
$enz->is_prototype;
}
=head2 Common REBASE parsing methods
The rest of the methods in this file are to be used by other REBASE
parsers. They are not to be used outside subclasses of this base
class. (They are 'protected' in the sense the word is used in Java.)
=cut
=head2 _cuts_from_site
Title : _cuts_from_site
Usage : ($site, $cut, $comp_cut) = _cuts_from_site('ACGCGT(4/5)');
Function: Separates cut positions from a single site string.
Does nothing to site if it does not have the cut string
Returns : array of site_string, forward_cut_position, reverse_cut_position
Args : recognition site string
Note : Not used in withrefm refactor/maj
=cut
sub _cuts_from_site {
my ($self, $site) = @_;
my ($cut, $comp_cut) = $site =~ /\((-?\d+)\/(-?\d+)\)/;
$site =~ s/\(.*\)$//;
return ($site, $cut, $comp_cut);
}
=head2 _meth
Title : _meth
Usage : ($pos, $meth) = $self->_meth('2(5)');
Function: Separates methylation postion and coce from a string.
Adjusts the postion depending on enzyme site length
and symmetry
Returns : array of position and methylation code
Args : 1. reference to Enzyme object
2. methylation description string
=cut
sub _meth {
my ($self, $re, $meth) = @_;
$meth =~ /(\S+)\((\d+)\)/;
my ($pos, $m) = ($1, $2);
$pos = 0 if $pos eq '?';
$pos = $re->seq->length + $pos if $pos and ! $re->palindromic;
return ($pos, $m);
$self->warn("Unknown methylation format [$meth]") if $self->verbose >0;
}
=head2 _coordinate_shift_to_cut
Title : _coordinate_shift_to_cut
Usage : $cut = $self->_coordinate_shift_to_cut($oricut, offset);
Function: Adjust cut position coordinates to start from the
first nucleotides of site
Returns : Cut position in correct coordinates
Args : 1. Original cut position
2. Length of the recognition site
Note : Not used in withrefm.pm refactor/maj
=cut
sub _coordinate_shift_to_cut {
my ($self, $cut, $site_length) = @_;
return $cut + $site_length;
}
=head2 _make_multisites
Title : _make_multisites
Usage : $self->_make_multisites($first_enzyme, \@sites, \@mets)
Function: Bless a Bio::Restriction::Enzyme into
Bio::Restriction::Enzyme::MultiSite and clone it as many
times as there are alternative sites.
Returns : nothing, does in place editing
Args : 1. a Bio::Restriction::Enzyme
2. reference to an array of recognition site strings
3. reference to an array of methylation code strings, optional
=cut
# removed the enzyme collection from arg list /maj
sub _make_multisites {
my ($self, $re, $sites, $meths, $xln_sub) = @_;
bless $re, 'Bio::Restriction::Enzyme::MultiSite';
my $count = 0;
while ($count < scalar @{$sites}) {
# this should probably be refactored to use the constructor
# too, rather than the clone/accessor method /maj
# my $re2 = $re->clone;
# my $re2;
my $site = @{$sites}[$count];
my ($precut, $recog, $postcut) = ( $site =~ m/^(?:\((\w+\/\w+)\))?([\w^]+)(?:\((\w+\/\w+)\))?/ );
# set the site attribute
# $re2->site($recog);
# set the recog attribute (which will make the regexp transformation
# if necessary:
# $re2->recog($recog);
# $recog = $re2->string;
# no warnings; # avoid 'uninitialized value' warning against $postcut
# my ($cut, $comp_cut) = ( $postcut =~ /(-?\d+)\/(-?\d+)/ );
# use warnings;
# note the following hard codes the coordinate transformation
# used for rebase/itype2 : this method will break on the
# base.pm format.
# if ($cut) {
# $re2->cut($cut + length $recog);
# $re2->complementary_cut($comp_cut + length $recog);
# }
my $re2 = Bio::Restriction::Enzyme::MultiSite->new(
-name => $re->name,
-site => $recog,
-recog => $recog,
-precut => $precut,
-postcut => $postcut,
-xln_sub => $xln_sub
);
if ($meths and @$meths) {
$re2->purge_methylation_sites;
$re2->methylation_sites($self->_meth($re2, @{$meths}[$count]));
}
$re->others($re2);
$count++;
}
foreach my $enz ($re->others) {
$enz->others($re, grep {$_ ne $enz} $re->others);
}
1;
}
=head2 _make_multicuts
Title : _make_multicuts
Usage : $self->_make_multicuts($first_enzyme, $precuts)
Function:
Bless a Bio::Restriction::Enzyme into
Bio::Restriction::Enzyme::MultiCut and clone it. The precut
string is processed to replase the cut sites in the cloned
object. Both objects refer to each other through others() method.
Returns : nothing, does in place editing
Args : 1. a Bio::Restriction::Enzyme
2. precut string, e.g. '12/7'
The examples we have of multiply cutting enzymes cut only four
times. This protected method deals only with a string of two
integers separated with a slash, e.g. '12/7'. The numbers represent the postions
BEFORE the start of the recognition site, i.e. negative positions.
=cut
# removed the enzyme collection from arg list /maj
sub _make_multicuts {
my ($self, $re, $precut) = @_;
bless $re, 'Bio::Restriction::Enzyme::MultiCut';
my ($cut, $comp_cut) = $precut =~ /(-?\d+)\/(-?\d+)/;
my $re2 = $re->clone;
$re2->cut("-$cut");
$re2->complementary_cut("-$comp_cut");
$re->others($re2);
1;
}
=head2 _companies
Title : _companies
Purpose : Defines the companies that we know about
Returns : A hash
Argument : Nothing
Comments : An internal method to define the companies that we know about
REBASE uses a code, and this converts the code to the real name
(e.g. A = Amersham Pharmacia Biotech)
=cut
sub _companies {
# this is just so it is easy to set up the codes that REBASE uses
my $self=shift;
my %companies=(
'A'=>'Amersham Pharmacia Biotech (1/03)',
'C'=>'Minotech Biotechnology (6/01)',
'E'=>'Stratagene (1/03)',
'F'=>'Fermentas AB (1/03)',
'G'=>'Qbiogene (1/03)',
'H'=>'American Allied Biochemical, Inc. (10/98)',
'I'=>'SibEnzyme Ltd. (1/03)',
'J'=>'Nippon Gene Co., Ltd. (6/00)',
'K'=>'Takara Shuzo Co. Ltd. (1/03)',
'M'=>'Roche Applied Science (1/03)',
'N'=>'New England Biolabs (1/03)',
'O'=>'Toyobo Biochemicals (11/98)',
'P'=>'Megabase Research Products (5/99)',
'Q'=>'CHIMERx (1/03)',
'R'=>'Promega Corporation (1/03)',
'S'=>'Sigma Chemical Corporation (1/03)',
'U'=>'Bangalore Genei (1/03)',
'V'=>'MRC-Holland (1/03)',
'X'=>'EURx Ltd. (1/03)');
$self->{company}=\%companies;
}
1;
__DATA__
AasI GACNNNNNNGTC 7
AatI AGGCCT 3
AccII CGCG 2
AatII GACGTC 5
AauI TGTACA 1
Acc113I AGTACT 3
Acc16I TGCGCA 3
Acc65I GGTACC 1
AccB1I GGYRCC 1
AccB7I CCANNNNNTGG 7
AccI GTMKAC 2
AccIII TCCGGA 1
AciI CCGC 1
AclI AACGTT 2
AcsI RAATTY 1
AcvI CACGTG 3
AcyI GRCGYC 2
AdeI CACNNNGTG 6
AfaI GTAC 2
AfeI AGCGCT 3
AflI GGWCC 1
AflII CTTAAG 1
AflIII ACRYGT 1
AgeI ACCGGT 1
AhaIII TTTAAA 3
AhdI GACNNNNNGTC 6
AhlI ACTAGT 1
AleI CACNNNNGTG 5
AluI AGCT 2
Alw21I GWGCWC 5
Alw44I GTGCAC 1
AlwNI CAGNNNCTG 6
Ama87I CYCGRG 1
AocI CCTNAGG 2
Aor51HI AGCGCT 3
ApaBI GCANNNNNTGC 8
ApaI GGGCCC 5
ApaLI GTGCAC 1
ApoI RAATTY 1
AscI GGCGCGCC 2
AseI ATTAAT 2
AsiAI ACCGGT 1
AsiSI GCGATCGC 5
AsnI ATTAAT 2
Asp700I GAANNNNTTC 5
Asp718I GGTACC 1
AspEI GACNNNNNGTC 6
AspHI GWGCWC 5
AspI GACNNNGTC 4
AspLEI GCGC 3
AspS9I GGNCC 1
AsuC2I CCSGG 2
AsuI GGNCC 1
AsuII TTCGAA 2
AsuNHI GCTAGC 1
AvaI CYCGRG 1
AvaII GGWCC 1
AviII TGCGCA 3
AvrII CCTAGG 1
AxyI CCTNAGG 2
BalI TGGCCA 3
BamHI GGATCC 1
BanI GGYRCC 1
BanII GRGCYC 5
BanIII ATCGAT 2
BbeI GGCGCC 5
BbrPI CACGTG 3
BbuI GCATGC 5
Bbv12I GWGCWC 5
BclI TGATCA 1
BcnI CCSGG 2
BcoI CYCGRG 1
BcuI ACTAGT 1
BetI WCCGGW 1
BfaI CTAG 1
BfmI CTRYAG 1
BfrBI ATGCAT 3
BfrI CTTAAG 1
BfuCI GATC 0
BglI GCCNNNNNGGC 7
BglII AGATCT 1
BlnI CCTAGG 1
BloHII CTGCAG 5
BlpI GCTNAGC 2
Bme1390I CCNGG 2
Bme1580I GKGCMC 5
Bme18I GGWCC 1
BmtI GCTAGC 5
BmyI GDGCHC 5
BoxI GACNNNNGTC 5
Bpu1102I GCTNAGC 2
Bpu14I TTCGAA 2
Bsa29I ATCGAT 2
BsaAI YACGTR 3
BsaBI GATNNNNATC 5
BsaHI GRCGYC 2
BsaJI CCNNGG 1
BsaOI CGRYCG 4
BsaWI WCCGGW 1
Bsc4I CCNNNNNNNGG 7
BscBI GGNNCC 3
BscFI GATC 0
BscI ATCGAT 2
Bse118I RCCGGY 1
Bse21I CCTNAGG 2
Bse8I GATNNNNATC 5
BseAI TCCGGA 1
BseBI CCWGG 2
BseCI ATCGAT 2
BseDI CCNNGG 1
BseJI GATNNNNATC 5
BseLI CCNNNNNNNGG 7
BsePI GCGCGC 1
BseSI GKGCMC 5
BseX3I CGGCCG 1
Bsh1236I CGCG 2
Bsh1285I CGRYCG 4
BshFI GGCC 2
BshI GGCC 2
BshNI GGYRCC 1
BshTI ACCGGT 1
BsiBI GATNNNNATC 5
BsiCI TTCGAA 2
BsiEI CGRYCG 4
BsiHKAI GWGCWC 5
BsiHKCI CYCGRG 1
BsiLI CCWGG 2
BsiMI TCCGGA 1
BsiQI TGATCA 1
BsiSI CCGG 1
BsiWI CGTACG 1
BsiXI ATCGAT 2
BsiYI CCNNNNNNNGG 7
BsiZI GGNCC 1
BslI CCNNNNNNNGG 7
BsoBI CYCGRG 1
Bsp106I ATCGAT 2
Bsp119I TTCGAA 2
Bsp120I GGGCCC 1
Bsp1286I GDGCHC 5
Bsp13I TCCGGA 1
Bsp1407I TGTACA 1
Bsp143I GATC 0
Bsp143II RGCGCY 5
Bsp1720I GCTNAGC 2
Bsp19I CCATGG 1
Bsp68I TCGCGA 3
BspA2I CCTAGG 1
BspCI CGATCG 4
BspDI ATCGAT 2
BspEI TCCGGA 1
BspHI TCATGA 1
BspLI GGNNCC 3
BspLU11I ACATGT 1
BspMII TCCGGA 1
BspT104I TTCGAA 2
BspT107I GGYRCC 1
BspTI CTTAAG 1
BspXI ATCGAT 2
BsrBRI GATNNNNATC 5
BsrFI RCCGGY 1
BsrGI TGTACA 1
BssAI RCCGGY 1
BssECI CCNNGG 1
BssHI CTCGAG 1
BssHII GCGCGC 1
BssKI CCNGG 0
BssNAI GTATAC 3
BssT1I CCWWGG 1
Bst1107I GTATAC 3
Bst2UI CCWGG 2
Bst4CI ACNGT 3
Bst98I CTTAAG 1
BstACI GRCGYC 2
BstAPI GCANNNNNTGC 7
BstBAI YACGTR 3
BstBI TTCGAA 2
BstC8I GCNNGC 3
BstDEI CTNAG 1
BstDSI CCRYGG 1
BstEII GGTNACC 1
BstENI CCTNNNNNAGG 5
BstENII GATC 0
BstFNI CGCG 2
BstH2I RGCGCY 5
BstHHI GCGC 3
BstHPI GTTAAC 3
BstKTI GATC 3
BstMAI CTGCAG 5
BstMCI CGRYCG 4
BstMWI GCNNNNNNNGC 7
BstNI CCWGG 2
BstNSI RCATGY 5
BstOI CCWGG 2
BstPAI GACNNNNGTC 5
BstPI GGTNACC 1
BstSCI CCNGG 0
BstSFI CTRYAG 1
BstSNI TACGTA 3
BstUI CGCG 2
BstX2I RGATCY 1
BstXI CCANNNNNNTGG 8
BstYI RGATCY 1
BstZ17I GTATAC 3
BstZI CGGCCG 1
Bsu15I ATCGAT 2
Bsu36I CCTNAGG 2
BsuRI GGCC 2
BsuTUI ATCGAT 2
BtgI CCRYGG 1
BthCI GCNGC 4
Cac8I GCNNGC 3
CaiI CAGNNNCTG 6
CauII CCSGG 2
CciNI GCGGCCGC 2
CelII GCTNAGC 2
CfoI GCGC 3
Cfr10I RCCGGY 1
Cfr13I GGNCC 1
Cfr42I CCGCGG 4
Cfr9I CCCGGG 1
CfrI YGGCCR 1
ChaI GATC 4
ClaI ATCGAT 2
CpoI CGGWCCG 2
Csp45I TTCGAA 2
Csp6I GTAC 1
CspAI ACCGGT 1
CspI CGGWCCG 2
CviAII CATG 1
CviJI RGCY 2
CviRI TGCA 2
CviTI RGCY 2
CvnI CCTNAGG 2
DdeI CTNAG 1
DpnI GATC 2
DpnII GATC 0
DraI TTTAAA 3
DraII RGGNCCY 2
DraIII CACNNNGTG 6
DrdI GACNNNNNNGTC 7
DsaI CCRYGG 1
DseDI GACNNNNNNGTC 7
EaeI YGGCCR 1
EagI CGGCCG 1
Eam1105I GACNNNNNGTC 6
Ecl136II GAGCTC 3
EclHKI GACNNNNNGTC 6
EclXI CGGCCG 1
Eco105I TACGTA 3
Eco130I CCWWGG 1
Eco147I AGGCCT 3
Eco24I GRGCYC 5
Eco32I GATATC 3
Eco47I GGWCC 1
Eco47III AGCGCT 3
Eco52I CGGCCG 1
Eco72I CACGTG 3
Eco81I CCTNAGG 2
Eco88I CYCGRG 1
Eco91I GGTNACC 1
EcoHI CCSGG 0
EcoICRI GAGCTC 3
EcoNI CCTNNNNNAGG 5
EcoO109I RGGNCCY 2
EcoO65I GGTNACC 1
EcoRI GAATTC 1
EcoRII CCWGG 0
EcoRV GATATC 3
EcoT14I CCWWGG 1
EcoT22I ATGCAT 5
EcoT38I GRGCYC 5
EgeI GGCGCC 3
EheI GGCGCC 3
ErhI CCWWGG 1
EsaBC3I TCGA 2
EspI GCTNAGC 2
FatI CATG 0
FauNDI CATATG 2
FbaI TGATCA 1
FblI GTMKAC 2
FmuI GGNCC 4
Fnu4HI GCNGC 2
FnuDII CGCG 2
FriOI GRGCYC 5
FseI GGCCGGCC 6
Fsp4HI GCNGC 2
FspAI RTGCGCAY 4
FspI TGCGCA 3
FunI AGCGCT 3
FunII GAATTC 1
HaeI WGGCCW 3
HaeII RGCGCY 5
HaeIII GGCC 2
HapII CCGG 1
HgiAI GWGCWC 5
HgiCI GGYRCC 1
HgiJII GRGCYC 5
HhaI GCGC 3
Hin1I GRCGYC 2
Hin6I GCGC 1
HinP1I GCGC 1
HincII GTYRAC 3
HindI CAC 2
HindII GTYRAC 3
HindIII AAGCTT 1
HinfI GANTC 1
HpaI GTTAAC 3
HpaII CCGG 1
Hpy178III TCNNGA 2
Hpy188I TCNGA 3
Hpy188III TCNNGA 2
Hpy8I GTNNAC 3
Hpy99I CGWCG 5
HpyCH4I CATG 3
HpyCH4III ACNGT 3
HpyCH4IV ACGT 1
HpyCH4V TGCA 2
HpyF10VI GCNNNNNNNGC 8
Hsp92I GRCGYC 2
Hsp92II CATG 4
HspAI GCGC 1
ItaI GCNGC 2
KasI GGCGCC 1
Kpn2I TCCGGA 1
KpnI GGTACC 5
Ksp22I TGATCA 1
KspAI GTTAAC 3
KspI CCGCGG 4
Kzo9I GATC 0
LpnI RGCGCY 3
LspI TTCGAA 2
MabI ACCWGGT 1
MaeI CTAG 1
MaeII ACGT 1
MaeIII GTNAC 0
MamI GATNNNNATC 5
MboI GATC 0
McrI CGRYCG 4
MfeI CAATTG 1
MflI RGATCY 1
MhlI GDGCHC 5
MlsI TGGCCA 3
MluI ACGCGT 1
MluNI TGGCCA 3
Mly113I GGCGCC 2
Mph1103I ATGCAT 5
MroI TCCGGA 1
MroNI GCCGGC 1
MroXI GAANNNNTTC 5
MscI TGGCCA 3
MseI TTAA 1
MslI CAYNNNNRTG 5
Msp20I TGGCCA 3
MspA1I CMGCKG 3
MspCI CTTAAG 1
MspI CCGG 1
MspR9I CCNGG 2
MssI GTTTAAAC 4
MstI TGCGCA 3
MunI CAATTG 1
MvaI CCWGG 2
MvnI CGCG 2
MwoI GCNNNNNNNGC 7
NaeI GCCGGC 3
NarI GGCGCC 2
NciI CCSGG 2
NcoI CCATGG 1
NdeI CATATG 2
NdeII GATC 0
NgoAIV GCCGGC 1
NgoMIV GCCGGC 1
NheI GCTAGC 1
NlaIII CATG 4
NlaIV GGNNCC 3
Nli3877I CYCGRG 5
NmuCI GTSAC 0
NotI GCGGCCGC 2
NruGI GACNNNNNGTC 6
NruI TCGCGA 3
NsbI TGCGCA 3
NsiI ATGCAT 5
NspBII CMGCKG 3
NspI RCATGY 5
NspIII CYCGRG 1
NspV TTCGAA 2
OliI CACNNNNGTG 5
PacI TTAATTAA 5
PaeI GCATGC 5
PaeR7I CTCGAG 1
PagI TCATGA 1
PalI GGCC 2
PauI GCGCGC 1
PceI AGGCCT 3
PciI ACATGT 1
PdiI GCCGGC 3
PdmI GAANNNNTTC 5
Pfl23II CGTACG 1
PflBI CCANNNNNTGG 7
PflFI GACNNNGTC 4
PflMI CCANNNNNTGG 7
PfoI TCCNGGA 1
PinAI ACCGGT 1
Ple19I CGATCG 4
PmaCI CACGTG 3
PmeI GTTTAAAC 4
PmlI CACGTG 3
Ppu10I ATGCAT 1
PpuMI RGGWCCY 2
PpuXI RGGWCCY 2
PshAI GACNNNNGTC 5
PshBI ATTAAT 2
PsiI TTATAA 3
Psp03I GGWCC 4
Psp124BI GAGCTC 5
Psp1406I AACGTT 2
Psp5II RGGWCCY 2
Psp6I CCWGG 0
PspAI CCCGGG 1
PspEI GGTNACC 1
PspGI CCWGG 0
PspLI CGTACG 1
PspN4I GGNNCC 3
PspOMI GGGCCC 1
PspPI GGNCC 1
PspPPI RGGWCCY 2
PssI RGGNCCY 5
PstI CTGCAG 5
PsuI RGATCY 1
PsyI GACNNNGTC 4
PvuI CGATCG 4
PvuII CAGCTG 3
RcaI TCATGA 1
RsaI GTAC 2
Rsr2I CGGWCCG 2
RsrII CGGWCCG 2
SacI GAGCTC 5
SacII CCGCGG 4
SalI GTCGAC 1
SanDI GGGWCCC 2
SatI GCNGC 2
Sau3AI GATC 0
Sau96I GGNCC 1
SauI CCTNAGG 2
SbfI CCTGCAGG 6
ScaI AGTACT 3
SciI CTCGAG 3
ScrFI CCNGG 2
SdaI CCTGCAGG 6
SduI GDGCHC 5
SecI CCNNGG 1
SelI CGCG 0
SexAI ACCWGGT 1
SfcI CTRYAG 1
SfeI CTRYAG 1
SfiI GGCCNNNNNGGCC 8
SfoI GGCGCC 3
Sfr274I CTCGAG 1
Sfr303I CCGCGG 4
SfuI TTCGAA 2
SgfI GCGATCGC 5
SgrAI CRCCGGYG 2
SgrBI CCGCGG 4
SinI GGWCC 1
SlaI CTCGAG 1
SmaI CCCGGG 3
SmiI ATTTAAAT 4
SmiMI CAYNNNNRTG 5
SmlI CTYRAG 1
SnaBI TACGTA 3
SpaHI GCATGC 5
SpeI ACTAGT 1
SphI GCATGC 5
SplI CGTACG 1
SrfI GCCCGGGC 4
Sse232I CGCCGGCG 2
Sse8387I CCTGCAGG 6
Sse8647I AGGWCCT 2
Sse9I AATT 0
SseBI AGGCCT 3
SspBI TGTACA 1
SspI AATATT 3
SstI GAGCTC 5
SstII CCGCGG 4
StuI AGGCCT 3
StyI CCWWGG 1
SunI CGTACG 1
SwaI ATTTAAAT 4
TaaI ACNGT 3
TaiI ACGT 4
TaqI TCGA 1
TasI AATT 0
TatI WGTACW 1
TauI GCSGC 4
TelI GACNNNGTC 4
TfiI GAWTC 1
ThaI CGCG 2
TliI CTCGAG 1
Tru1I TTAA 1
Tru9I TTAA 1
TscI ACGT 4
TseI GCWGC 1
Tsp45I GTSAC 0
Tsp4CI ACNGT 3
Tsp509I AATT 0
TspEI AATT 0
Tth111I GACNNNGTC 4
TthHB8I TCGA 1
UnbI GGNCC 0
Van91I CCANNNNNTGG 7
Vha464I CTTAAG 1
VneI GTGCAC 1
VpaK11AI GGWCC 0
VpaK11BI GGWCC 1
VspI ATTAAT 2
XagI CCTNNNNNAGG 5
XapI RAATTY 1
XbaI TCTAGA 1
XceI RCATGY 5
XcmI CCANNNNNNNNNTGG 8
XhoI CTCGAG 1
XhoII RGATCY 1
XmaCI CCCGGG 1
XmaI CCCGGG 1
XmaIII CGGCCG 1
XmaJI CCTAGG 1
XmiI GTMKAC 2
XmnI GAANNNNTTC 5
XspI CTAG 1
ZhoI ATCGAT 2
ZraI GACGTC 3
Zsp2I ATGCAT 5