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#!/usr/bin/env ruby
require 'optparse'
require 'ostruct'
# Add code to check whether a class is already defined
def class_exists?(class_name)
klass = Module.const_get(class_name)
return klass.is_a?(Class)
rescue NameError
return false
end
# Ruby < 1.9.2 does not have the Random class.
# Add a small substitute that does something similar.
if !(class_exists?('Random')) then
class Random
def initialize(seed = nil)
if seed.nil? then
seed = self.new_seed
end
@seed = seed
Kernel.srand(seed)
end
def self.new_seed
return Kernel.srand()
end
def rand(vmax = nil)
if vmax.nil? then
return Kernel.rand()
else
return (vmax * Kernel.rand()).to_i
end
end
end
end
class String
def reverse_complement
return self.reverse.tr("ACGTacgt","TGCAtgca")
end
end
class Randomsequence
def initialize(alphabet,myseed=nil)
@alphabet = alphabet
@asize = alphabet.length
if myseed.nil?
seed = Random.new_seed
else
seed = myseed
end
STDERR.puts "seed=#{seed}"
@rgen = Random.new(seed)
end
def sequence(len)
s = Array.new()
0.upto(len-1).each do
s.push(@alphabet[@rgen.rand * @asize, 1])
end
return s.join
end
def mutate(sequence,errperc,alphabet)
len = sequence.length
err_prob = errperc.to_f/100.0
asize = alphabet.length
s = Array.new()
i = 0
loop do
r = @rgen.rand
if r <= err_prob
r = @rgen.rand
if r <= 0.8
s.push(alphabet[@rgen.rand * asize, 1])
i += 1
elsif r <= 0.9
s.push(alphabet[@rgen.rand * asize, 1])
else
i += 1
end
else
s.push(sequence[i, 1])
i += 1
end
if i >= len
break
end
end
return s.join("")
end
def rgen()
return @rgen
end
end
def parseargs(argv)
options = OpenStruct.new
mode = nil
minminid = 70
maxminid = 99
defaultminid = 80
indent = 37
minidrange = "[#{minminid}..#{maxminid}]"
options.seedlength = nil
options.totallength = nil
options.pairparam = false
options.mirrored = false
options.seeded = false
options.namedfiles = false
options.withwildcards = false
options.minidentity = defaultminid
options.seednumber = Random.new_seed
options.reverse = false
options.reverse_complement = false
options.number = 1
options.mems = false
options.seedcoverage = 0
options.long = 0
opts = OptionParser.new
opts.on("-m","--mode STRING","specify mode: mirrored|seeded|pair") do |x|
mode = x
end
opts.on("-s","--seedlength NUM","specify seed length for mirrored sequences") do |x|
options.seedlength = x.to_i
end
opts.on("-l","--length NUM","specify total length of sequences") do |x|
options.totallength = x.to_i
end
opts.on("-i","--minidentity NUM","specify minimum identity percentage in\n" +
(" " * indent) + "range " + "#{minidrange}, " +
"default is #{defaultminid}") do |x|
options.minidentity = x.to_i
end
opts.on("-n","--namedfiles","store seeded matches in files db.fna and\n" +
(" " * indent) + "query.fna") do |x|
options.namedfiles = true
end
opts.on("-w","--withwildcards","store wildcards at end of extensions") do |x|
options.withwildcards = true
end
opts.on("-r","--reverse","reverse the query sequence in query of\n" +
(" " * indent) + "seeded sequence pair") do |x|
options.reverse = true
end
opts.on("-p","--reverse-complement","reverse complement the query \n" +
(" " * indent) +
"sequence in query of seeded sequence pair") do |x|
options.reverse_complement = true
end
opts.on("--mems","generate mems, i.e. the seed are maximal") do |x|
options.mems = true
end
opts.on("--number NUM","specify the number of seeded pairs to be\n" +
(" " * indent) + "generated (default 1)") do |x|
options.number = x.to_i
end
opts.on("--seed NUM","specify the seed for the random number\n" +
(" " * indent) + "generator to make sequences reproducible") do |x|
options.seednumber = x.to_i
end
opts.on("--long NUM","generate a long sequence with multiple\n" + (" " * indent) +
"seeds and noisy areas in between") do |x|
options.long = x.to_i
end
opts.on("-c","--seedcoverage NUM","generate multiple seeds that cover the\n" +
(" " * indent) + "specified number of bases") do |x|
options.seedcoverage = x.to_i
end
rest = opts.parse(argv)
if rest.length != 0
STDERR.puts "Usage: #{$0} [options]"
exit 1
end
if mode.nil?
STDERR.puts "#{$0}: option -m is mandatory"
exit 1
end
if mode == "mirrored"
options.mirrored = true
elsif mode == "seeded"
options.seeded = true
elsif mode == "pair"
options.pairparam = true
else
STDERR.puts "#{$0}: possible modes are mirrored|seeded|pair"
exit 1
end
if not (options.seeded or options.mirrored)
if not options.seedlength.nil?
STDERR.puts "#{$0}: option -s requires option -m seeded or -m mirrored"
exit 1
end
if options.namedfiles
STDERR.puts "#{$0}: option -n requires option -m seeded or -m mirrored"
exit 1
end
end
if options.totallength.nil?
STDERR.puts "#{$0}: option --length is mandatory"
exit 1
end
if (options.mirrored or options.seeded) and options.seedlength.nil?
STDERR.puts "#{$0}: option -mode mirrored and -m seeded imply option -s"
exit 1
end
if options.seedcoverage != 0 and not options.seeded
STDERR.puts "#{$0}: option -c requires option -m seeded"
exit 1
end
if options.long != 0 and options.seedcoverage == 0
STDERR.puts "#{$0}: option --long requires option -c"
exit 1
end
if not options.seedlength.nil? and options.seedlength >= options.totallength
STDERR.puts "#{$0}: seedlength must not be larger than totallength"
exit 1
end
if options.minidentity < minminid or options.minidentity > maxminid
STDERR.puts "#{$0}: minidentity must be in range #{minidrange}"
exit 1
end
if options.seedcoverage >= options.totallength
STDERR.puts "#{$0}: seedcoverage must not be larger than totallength"
exit 1
end
if options.reverse and options.reverse_complement
STDERR.puts "#{$0}: options -reverse and -reverse_complement exclude each other"
exit 1
end
return options
end
def gen_mirrored(fpdb,fpquery,rseq,options,alphabet,errperc)
extendlength = (options.totallength - options.seedlength)/2
seedstring = rseq.sequence(options.seedlength)
leftcontext1 = rseq.sequence(extendlength)
leftcontext2 = rseq.mutate(leftcontext1,errperc,alphabet)
fpdb.puts ">seedlength=#{options.seedlength},extendlength=#{extendlength}," +
"errperc=#{errperc}"
fpdb.puts "#{leftcontext1}"
fpdb.puts "#{seedstring}"
fpdb.puts "#{leftcontext1.reverse}"
fpquery.puts ">seedlength=#{options.seedlength},extendlength=#{extendlength}," +
"errperc=#{errperc}"
fpquery.puts "#{leftcontext2}"
fpquery.puts "#{seedstring}"
fpquery.puts "#{leftcontext2.reverse}"
end
# headkey with 1 seed
def headkey(options,extendlength,errperc,seedpos)
return "seedlength=#{options.seedlength},extendlength=#{extendlength}," +
"errperc=#{errperc},seedpos=#{seedpos}..#{seedpos+options.seedlength-1}"
end
# headkey with multiple seeds (-c option)
def headkey_cov(options,extendlength,errperc)
return "seedlength=#{options.seedlength},sequencelength=#{extendlength}," +
"errperc=#{errperc}"
end
def namekey(b)
if b
return "r"
else
return "p"
end
end
# generate 1 seed in the middle of a sequence
def gen_seeded(fpdb,fpquery,fpquery_r,rseq,options,alphabet,errperc)
extendlength = (options.totallength - options.seedlength)/2
seedstring = rseq.sequence(options.seedlength)
leftcontext1 = rseq.sequence(extendlength)
leftcontext2 = rseq.mutate(leftcontext1,errperc,alphabet)
rightcontext1 = rseq.sequence(extendlength)
rightcontext2 = rseq.mutate(rightcontext1,errperc,alphabet)
if options.withwildcards
wildcard = "N"
else
wildcard = ""
end
if options.mems
left1 = "a"
left2 = "c"
right1 = "g"
right2 = "t"
else
left1 = ""
left2 = ""
right1 = ""
right2 = ""
end
seedpos="#{leftcontext1}#{wildcard}#{left1}".length
fpdb.puts ">db: #{headkey(options,extendlength,errperc,seedpos)}"
fpdb.puts "#{leftcontext1}#{wildcard}#{left1}"
fpdb.puts "#{seedstring}"
fpdb.puts "#{right1}#{rightcontext1}"
seedpos="#{leftcontext2}#{left2}".length
fpquery.puts ">query: #{headkey(options,extendlength,errperc,seedpos)}"
queryseq = "#{leftcontext2}#{left2}\n#{seedstring}\n#{right2}#{rightcontext2}"
fpquery.puts queryseq
if options.reverse or options.reverse_complement
seedpos="#{right2}#{rightcontext2}".length
key = namekey(options.reverse)
fpquery_r.puts ">query-#{key}: #{headkey(options,extendlength,errperc,seedpos)}"
if options.reverse
fpquery_r.puts queryseq.reverse
else
fpquery_r.puts queryseq.reverse_complement
end
end
end
# distribute several seeds over a sequence
def gen_seeded_with_coverage(rseq,options,alphabet,errperc,dbseq,queryseq,
seedpos)
# build array with random seed positions covering >= seedcoverage positions
pos = Array.new
while pos.length < options.seedcoverage
idx = rseq.rgen.rand * (options.totallength - options.seedlength + 1)
idx = idx.to_i
pos.concat((idx...idx + options.seedlength).to_a).sort!.uniq!
end
# convert array to ranges
ranges = pos.inject([]) do |spans, n|
if spans.empty? || spans.last.last != n - 1
spans + [n..n]
else
spans[0..-2] + [spans.last.first..n]
end
end
# generate sequences
context1 = Array.new
seedstring = Array.new
prev = 0
for range in ranges
context1.push("#{rseq.sequence(range.begin - prev)}")
range_size = range.last - range.first + 1
seedstring.push("#{rseq.sequence(range_size)}")
prev = range.last + 1
end
context1.push("#{rseq.sequence(options.totallength - prev)}")
context2 = context1.map{|sequence| rseq.mutate(sequence, errperc, alphabet)}
if options.withwildcards
wildcard = "N"
else
wildcard = ""
end
if options.mems
left1 = "a"
left2 = "c"
right1 = "g"
right2 = "t"
else
left1 = ""
left2 = ""
right1 = ""
right2 = ""
end
# extract sequences
for i in (0...seedstring.length)
dbseq += "#{context1[i]}#{wildcard}#{left1}"
seedpos[0].push((dbseq.length..dbseq.length+seedstring[i].length-1)) unless seedpos.nil?
dbseq += "#{seedstring[i]}#{right1}"
end
dbseq += "#{context1[seedstring.length]}"
dbhead = ">db: #{headkey_cov(options,dbseq.length,errperc)}"
for i in (0...seedstring.length)
queryseq += "#{context2[i]}#{left2}"
seedpos[1].push((queryseq.length..queryseq.length+seedstring[i].length-1)) unless seedpos.nil?
queryseq += "#{seedstring[i]}#{right2}"
end
queryseq += "#{context2[seedstring.length]}"
queryhead = ">query: #{headkey_cov(options,queryseq.length,errperc)}"
if options.reverse or options.reverse_complement
key = namekey(options.reverse)
query_r = ">query-#{key}: #{headkey_cov(options,queryseq.length,errperc)}"
else
query_r = nil
end
return dbhead, dbseq, queryhead, queryseq, query_r, seedpos
end
# create 1 long sequence with several alignment regions and noise in between
def gen_long_seeded(rseq,options,alphabet,errperc,dbinit,queryinit,seedinit)
dbseq = dbinit + rseq.sequence((rseq.rgen.rand * 50).to_i)
queryseq = queryinit + rseq.sequence((rseq.rgen.rand * 50).to_i)
seedpos = seedinit
while dbseq.length < options.long
data = gen_seeded_with_coverage(rseq,options,alphabet,errperc,dbseq,
queryseq,nil)
seedpos[0].push((dbseq.length+1..data[1].length))
seedpos[1].push((queryseq.length+1..data[3].length))
dbseq = data[1] + rseq.sequence((rseq.rgen.rand * 300 + 100).to_i)
queryseq = data[3] + rseq.sequence((rseq.rgen.rand * 300 + 100).to_i)
end
dbhead = ">db: #{headkey_cov(options,dbseq.length,errperc)}"
queryhead = ">query: #{headkey_cov(options,queryseq.length,errperc)}"
return data[0], dbseq, data[2], queryseq, data[4], seedpos
end
def seq_to_fp(fpdb,fpquery,fpquery_r,data)
# extract seed positions to string format
seedpos = data[5][0]
strseedpos = seedpos.map{ |range| "#{range.first}..#{range.last}"}.join("|")
# write db header and sequence
fpdb.puts data[0] + ",seedpos=" + strseedpos
dbseq = data[1]
seedpos.reverse_each{ |range|
dbseq.insert(range.last+1,"\n")
dbseq.insert(range.first,"\n")
}
fpdb.puts dbseq
# write query header and sequence
seedpos = data[5][1]
strseedpos = seedpos.map{ |range| "#{range.first}..#{range.last}"}.join("|")
fpquery.puts data[2] + ",seedpos=" + strseedpos
if not data[4].nil? then
strseedpos = seedpos.reverse.map{ |range|
"#{data[3].length-range.last-1}..#{data[3].length-range.first-1}"
}.join("|")
end
queryseq = data[3]
seedpos.reverse_each{ |range|
queryseq.insert(range.last+1,"\n")
queryseq.insert(range.first,"\n")
}
fpquery.puts queryseq
# write reverse query
if not data[4].nil? then
fpquery_r.puts data[4] + ",seedpos=" + strseedpos
fpquery_r.puts queryseq.reverse
end
end
def openoutfile(filename)
begin
fp = File.new(filename,"w")
rescue => err
STDERR.puts "cannot open #{filename}"
exit 1
end
return fp
end
options = parseargs(ARGV)
alphabet = "acgt"
errperc = 100 - options.minidentity
rseq = Randomsequence.new(alphabet,options.seednumber)
if options.namedfiles
fpdb = openoutfile("db.fna")
fpquery = openoutfile("query.fna")
if options.reverse or options.reverse_complement
key = namekey(options.reverse)
fpquery_r = openoutfile("query-#{key}.fna")
else
fpquery_r = nil
end
else
fpdb = STDOUT
fpquery = STDOUT
fpquery_r = STDOUT
end
if options.mirrored
gen_mirrored(fpdb,fpquery,rseq,options,alphabet,errperc)
elsif options.seeded
options.number.times do
if options.seedcoverage == 0
gen_seeded(fpdb,fpquery,fpquery_r,rseq,options,alphabet,errperc)
else
seedpos = Array.new(2){Array.new}
dbseq = String.new
queryseq = String.new
if options.long != 0
data = gen_long_seeded(rseq,options,alphabet,errperc,dbseq,queryseq,
seedpos)
else
data = gen_seeded_with_coverage(rseq,options,alphabet,errperc,dbseq,
queryseq,seedpos)
end
seq_to_fp(fpdb,fpquery,fpquery_r,data)
end
end
elsif options.pairparam
seq1 = rseq.sequence(options.totallength)
seq2 = rseq.sequence(options.totallength)
puts "#{seq1} #{seq2}"
end
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