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#!/usr/bin/python
import os, sys, argparse, logging, json
from tempfile import NamedTemporaryFile
from collections import namedtuple
from pbsuite.utils.setupLogging import setupLogging
from pbsuite.utils.FileHandlers import FastqFile, M5File, M4File, revComp
from pbsuite.jelly.Support import AlignmentConnector, SUPPORTFLAGS
from pbsuite.banana.Polish import *
import pbsuite.jelly.m4pie as m4pie
ALLTEMPFILES = []
MINTAIL = 200
GAPWIGGLE = 400 # max deviation from gapsize a span seq's fill can be
def blasr(query, target, fmt="5", bestn=20, nCandidates=20, nproc = 1, outname = "out.m5"):
"""
Simple overlapper
"""
c = ("blasr %s %s -m %s --bestn %d --nCandidates %d --minMatch 8 --sdpTupleSize 6 --affineAlign "
"--nproc %d --noSplitSubreads --out %s --minPctIdentity 60 --minReadLength 5") % \
(query, target, fmt, bestn, nCandidates, nproc, outname)
logging.debug(c)
r,o,e = exe(c)
logging.debug("blasr - %d - %s - %s" % (r, o, e))
def tailblasr(query, target, nproc=1, outname="out.m5", basedir="./"):
"""
Try getting the read to hit each target uniquely instead of hoping that bestn reports all possible alignments
"""
global ALLTEMPFILES
#input reads
reads = FastqFile(query)
#map to make the primary
primary= NamedTemporaryFile(prefix="primary_", suffix=".m4", delete=False, dir=basedir)
primary = primary.name
ALLTEMPFILES.append(primary)
blasr(query, target, fmt="4", nproc=nproc, bestn=1, outname=primary)
#build command to call m4pie
args = "%s %s %s -t %d -n %d -o %s" % (primary, query, target, MINTAIL, nproc, outname)
args = args.split()
m4pie.run(args)
def oldtails():
aligns = M5File(primary)
#where I'm putting the good hits
mapOut = open(outname, "w")
#where I'm putting the tails
tfq = NamedTemporaryFile(prefix="tails_", suffix=".fastq", delete=False, dir=basedir)
ALLTEMPFILES.append( tfq.name )
whichEnd = defaultdict(list)
#extract the tails
ntails = 0
for a in aligns:
if a.qstart >= MINTAIL:
tseq1 = reads[a.qname].subSeq(None, a.qstart)
#prolog
tseq1.name = "%s_::_5_::_%d,%d" % (tseq1.name, a.qstart, a.qseqlength)
tfq.write(str(tseq1))
ntails += 1
if a.qend - a.qseqlength > MINTAIL:
tseq2 = reads[a.qname].subSeq(a.qend, None)
#epilog
tseq2.name = "%s_::_3_::_%d,%d" % (tseq2.name, a.qend, a.qseqlength)
tfq.write(str(tseq2))
ntails += 1
mapOut.write(str(a)+"\n")
#don't want redundant hits on a single flank
whichEnd[a.qname].append(a.tname)
tfq.close()
logging.info("%d unmapped tails" % (ntails))
#map tails
tailAlign = NamedTemporaryFile(prefix="tails_", suffix=".m5", delete=False, dir=basedir)
tailAlign = tailAlign.name
ALLTEMPFILES.append(tailAlign)
blasr(tfq.name, target, nproc=nproc, bestn=1, outname=tailAlign)
aligns2 = M5File(tailAlign)
logging.info("%d tails mapped" % len(aligns2))
for a in aligns2:
#get the carryon info
name, direct, se = a.qname.split("_::_")
pos, length = map(int, se.split(','))
#correct it's information
a.qname = name
a.qseqlength = length
#prevent redundant flank map
if a.tname in whichEnd[a.qname]:
logging.info("%s failed ref map" % a.tname)
continue
whichEnd[a.qname].append(a.tname)
#epilogs need to be updated
if direct == '3':
a.qstart += pos
a.qend += pos
mapOut.write(str(a)+"\n")
mapOut.close()
return
def extractFlanks(reads, basedir="./"):
"""
Takes FastqFile and separates the the reference reads (^ref)
from the supporting reads
returns queryFileName, targetFileName
"""
global ALLTEMPFILES
query = NamedTemporaryFile(prefix="query_", suffix=".fastq", delete=False, dir=basedir)
ALLTEMPFILES.append(query.name)
target = NamedTemporaryFile(prefix="target_", suffix=".fasta", delete=False, dir=basedir)
ALLTEMPFILES.append(target.name)
for read in reads:
if read.startswith("ref"):
target.write(">%s\n%s\n" % (read, reads[read].seq))
else:
query.write(reads[read].toString())
query.close()
target.close()
return query.name, target.name
def orderSeeds(seedNames):
"""
Looks at the seed's names to figure out
which one is upstream of the next and if alignments
should be on the same strand
"""
if len(seedNames) == 1:
seedNames.append(None)
return True, seedNames
seed1, seed2 = seedNames
logging.debug("Ordering %s and %s" % (seed1, seed2))
if seed1 == None:
logging.error("Seed1 must be non-None to AssessAssembly!")
exit(5)
#I will be returning a None, just need to know
#if seed1 is trying to extend 5' or 3'
if seed2 == None:
sameStrand = True
if seed1.endswith("e3"):
ret = (None, seed1)
elif seed1.endswith("e5"):
ret = (seed1, None)
elif seed1.endswith("e3") and seed2.endswith("e5"):
sameStrand = True
ret = (seed1, seed2)
elif seed1.endswith("e5") and seed2.endswith("e3"):
sameStrand = True
ret = (seed2, seed1)
else:
#if seed1.endswith("e5") and seed2.endswith("e5"):
#if seed1.endswith("e3") and seed2.endswith("e3"):
#No way to know. Someone is reverse-compliment of
#the other. -- One needs to be on the opposite Strand
sameStrand = False
ret = (seed1, seed2)
logging.debug(("Seed Order %s - %s : strand -" % ret) + \
str(sameStrand))
return sameStrand, ret
def createStats():
"""
I just wanted to separate the stats so It is a little cleaner
"""
#span, seed1, seed2
return {"support": [[], [], []], #keep all the flags I have \
"spanCount": 0,
"spanSeedName": None,
"spanSeedScore": 0,
"spanSeedStart": None,
"spanSeedEnd": None,
"spanSeedStrand1": None,
"spanSeedStrand2": None,
"avgSpanBases": 0,
"seed1": None,
"seed2": None,
"predictedGapSize": None,
"sameStrand": None,
"extendF1Count": 0,
"avgExtF1Bases": 0,
"extendF1SeedName": 0,
"extendF1SeedScore": 0,
"extendF1SeedStart": None,
"extendF1SeedEnd": None,
"extendF1SeedStrand": None,
"extendF2Count": 0,
"avgExtF2Bases": 0,
"extendF2SeedName": 0,
"extendF2SeedScore": 0,
"extendF2SeedStart": None,
"extendF2SeedEnd": None,
"extendF2SeedStrand": None,
"extendSeq1": None,
"extendSeq2": None,
"fillSeq": None,
"contribSeqs": 0,
"contribBases": 0,
"fillBases": 0,
"seed1Trim": 0,
"seed2Trim": 0}
def getSubSeqs(alignmentFile, readsFile, sameStrand, seeds, predictedGapSize, maxTrim, maxWiggle, basedir="./"):
"""
Finds the seqs that align to the flanks the best, creates a fastq of supporting reads
and the seed
Might have a problem with my best read no going off the edge fully
so I put the maxFlank at 20
I should do more strand correction here
"""
global ALLTEMPFILES
def singleExtendLookup(sup, a):
"""
For getting how a single read extends a single flank
"""
if sup == SUPPORTFLAGS.none:
return None
#set the coordinates of the extending sequence
logging.debug(sup)
logging.debug(a.qname)
mystart = None
myend = None
if a.tname.endswith("e5") and sup in [SUPPORTFLAGS.left, SUPPORTFLAGS.span]:
if a.tstrand == '0':
mystart = 0
myend = a.qstart
else:
mystart = a.qend
myend = a.qseqlength
elif a.tname.endswith("e3") and sup in [SUPPORTFLAGS.right, SUPPORTFLAGS.span]:
if a.tstrand == '0':
mystart = a.qend
myend = a.qseqlength
else:
mystart = 0
myend = a.qstart
if mystart is None or myend is None or mystart < 0 or myend > a.qseqlength:
return None
#tscore = a.score * (myend - mystart)
#what flank and is it the best
if a.tname.replace('/','.') == stats["seed1"]:
stats["extendF1Count"] += 1
stats["avgExtF1Bases"] += a.qstart
stats["support"][1].append( sup )
if a.score < stats["extendF1SeedScore"]:
stats["extendF1SeedScore"] = a.score #tscore
stats["extendF1SeedName"] = a.qname
stats["extendF1SeedStart"] = mystart
stats["extendF1SeedEnd"] = myend
stats["extendF1SeedStrand"] = a.tstrand
return reads[a.qname].subSeq(mystart, myend)
#myOut = f1fout
elif a.tname.replace('/','.') == stats["seed2"]:
stats["extendF2Count"] += 1
stats["avgExtF2Bases"] += a.qstart
stats["support"][2].append( sup )
if a.score < stats["extendF2SeedScore"]:
stats["extendF2SeedScore"] = a.score#tscore
stats["extendF2SeedName"] = a.qname
stats["extendF2SeedStart"] = mystart
stats["extendF2SeedEnd"] = myend
stats["extendF2SeedStrand"] = a.tstrand
return reads[a.qname].subSeq(mystart, myend)
#myOut = f2fout
#myOut.write(str(reads[a.qname].subSeq(mystart, myend)))
return None
connector = AlignmentConnector()
#aligns = connector.parseAlignments(M5File(alignmentFile))
#no need to connect with the tailmap
aligns = defaultdict(list)
for a in M4File(alignmentFile):
aligns[a.qname].append(a)
aligns = aligns.values()
reads = FastqFile(readsFile)
stats = createStats()
stats["seed1"], stats["seed2"] = seeds
stats["sameStrand"] = sameStrand
bestSpan = None
bestF1E = None
bestF2E = None
for readGroup in aligns:
if len(readGroup) > 2:
best = 0
worst = 0
keep = []
for i in readGroup:
if i.score < best:
keep.insert(0, i)
if len(keep) >= 2:
keep.pop()
best = i.score
elif i.score < worst:
keep.insert(1,i)
if len(keep) >=2:
keep.pop()
worst = i.score
readGroup = keep
if len(readGroup) == 2:
#make sure that the two hits aren't hitting the same target
if readGroup[0].tname == readGroup[1].tname:
if readGroup[0].score <= readGroup[1].score:
del(readGroup[1])
else:
del(readGroup[0])
#hit on each flank
if len(readGroup) == 2:
r1, r2 = readGroup
if r1.tname == stats["seed2"]:
r1, r2 = r2, r1
a = connector.extendsTarget(r1, maxFlank=maxTrim, minCovers=0)
logging.debug(a)
#Also check appropriate orientation
if r1.tname.endswith('e3'):
if a not in [SUPPORTFLAGS.right, SUPPORTFLAGS.span]:
logging.debug('reset a')
a = SUPPORTFLAGS.none
elif r1.tname.endswith('e5'):
if a not in [SUPPORTFLAGS.left, SUPPORTFLAGS.span]:
logging.debug('reset a')
a = SUPPORTFLAGS.none
b = connector.extendsTarget(r2, maxFlank=maxTrim, minCovers=0)
if r2.tname.endswith('e3'):
if b not in [SUPPORTFLAGS.right, SUPPORTFLAGS.span]:
logging.debug('reset b')
b = SUPPORTFLAGS.none
elif r2.tname.endswith('e5'):
if b not in [SUPPORTFLAGS.left, SUPPORTFLAGS.span]:
logging.debug('reset b')
b = SUPPORTFLAGS.none
elif len(readGroup) == 1:
r1 = readGroup[0]
r2 = None
a = connector.extendsTarget(r1, maxFlank=10)
b = SUPPORTFLAGS.none
if r1.tname == stats["seed2"]:
r1, r2 = r2, r1
a, b = b, a
else:
logging.warning("read %s gave too many alignments" % (readGroup[0].qname))
#it extends both flanks
if a != SUPPORTFLAGS.none and b != SUPPORTFLAGS.none:
logging.debug("%s spans" % r1.qname)
logging.debug("aflag %d bflag %d" % (a,b))
logging.debug("hit1- %s (%d, %d)" % (r1.tname, r1.qstart, r1.qend))
logging.debug("hit2- %s (%d, %d)" % (r2.tname, r2.qstart, r2.qend))
rStart = min(r1.qend, r2.qend)
rEnd = max(r1.qstart, r2.qstart)
sz = rEnd - rStart
tooShort = False
if sz < 50:
logging.info("fill seq is too short to call consensus")
tooShort = True
tooShortSeq = reads[r1.qname].subSeq(rStart, rEnd)
#continue
if predictedGapSize is not None and (predictedGapSize - sz) > maxWiggle:
logging.info("fill seq size %d is smaller than allowed predicted gap size wiggle %d" % (sz, maxWiggle))
continue
#Need to ensure that it's extending in the correct orientation
#need to ensure that everything is on the right strand
if sameStrand and r1.tstrand != r2.tstrand:
logging.debug("bad strandedness")
continue
#check for negative gaps
stats["spanCount"] += 1
stats["avgSpanBases"] += rEnd - rStart
stats["support"][0].append(SUPPORTFLAGS.span)
t = reads[r1.qname].subSeq(rStart, rEnd)
#sfout.write(str(t))
#is it the best spanner
score = r1.score + r2.score
if score < stats["spanSeedScore"]:
logging.debug("scoring %s %s" % (r1.qname, r2.qname))
stats["spanSeedScore"] = score
spanSeedName = r1.qname
stats["spanSeedStrand1"] = r1.tstrand
bestSpan = reads[r1.qname].subSeq(rStart, rEnd)
stats["spanSeedName"] = r1.qname
stats["spanSeedStart"] = rStart
stats["spanSeedEnd"] = rEnd
stats["spanSeedStrand2"] = r2.tstrand
stats["spanShort"] = tooShort
if r1.tname.endswith('e5'):
stats["seed1Trim"] = r1.tstart
logging.debug('trim1 %d' % (r1.tstart))
else:
stats["seed1Trim"] = r1.tseqlength - r1.tend
logging.debug('trim1else %d' % (r1.tseqlength - r1.tend))
if r2.tname.endswith('e5'):
stats["seed2Trim"] = r2.tstart
logging.debug('trim2 %d' % (r2.tstart))
else:
stats["seed2Trim"] = r2.tseqlength - r2.tend
logging.debug('trimelse %d' % (r2.tseqlength - r2.tend))
c = singleExtendLookup(a, r1)
if c is not None:
bestF1E = c
c = singleExtendLookup(b, r2)
if c is not None:
bestF2E = c
#sfout.close()
#sfout = sfout.name
#f1fout.close()
#f1fout = f1fout.name
#f2fout.close()
#f2fout = f2fout.name
logging.info("%d reads span" % stats["spanCount"])
logging.info("%d reads extend flank 1" % stats["extendF1Count"])
logging.info("%d reads extend flank 2" % stats["extendF2Count"])
#nt = namedtuple("SubInfo", "stats spanReads flank1Reads flank2Reads spanSeed flank1Seed flank2Seed")
nt = namedtuple("SubInfo", "stats spanSeed flank1Seed flank2Seed")
#seeds out files
ssfout = None
f1sfout = None
f2sfout = None
#replace too short with N's
#if stats["spanCount"] == 0 and len(tooShort) > (stats["extendF1Count"] + stats["extendF2Count"])/2:
"""This is when I would say "oh, i'm too short - and stop early. Now, I'm still going to try to write the
short stuff and treat it like anything else. It'll be up to later processes to catch this guy.
if stats["spanCount"] != 0 and stats["spanShort"]:
#stats["avgSpanBases"] =
#stats["spanCount"] = len(tooShort)
logging.info("estimated fill len %d" % (stats["avgSpanBases"]))
logging.debug("but I'm too short")
#stats["fillSeq"] = "N"* abs(stats["spanSeedStart"] - stats["spanSeedEnd"])
stats["fillSeq"] = tooShortSeq
stats["spanSeedScore"] = -500
stats["spanSeedStrand1"] = '0'
stats["spanSeedStrand2"] = '0'
#stats["spanSeedName"] = "tooShortNs"
#ret = nt(stats, None, None, None, None, None, None)
ret = nt(stats, None, None, None)
return ret
"""
if stats["spanCount"] > 0:
stats["avgSpanBases"] = stats["avgSpanBases"]/stats["spanCount"]
logging.info("estimated fill len %d" % (stats["avgSpanBases"]))
#write seed
if len(bestSpan.seq) < 50:
logging.warning("fill sequence is small (%dbp) can't call consensus" % (len(bestSpan.seq)))
#I don't know what to return here
ssfout = NamedTemporaryFile(prefix="span_", suffix=".fasta", delete=False, dir=basedir)
ALLTEMPFILES.append(ssfout.name)
logging.debug("spanning with %s" % (bestSpan.name))
ssfout.write(">%s\n%s\n" % (bestSpan.name, bestSpan.seq))
ssfout.close()
ssfout = ssfout.name
#if stats["extendF1Count"] > 0:
if bestF1E is not None:
stats["avgExtF1Bases"] = stats["avgExtF1Bases"]/stats["extendF1Count"]
logging.info("estimated flank 1 extend len %d" % (stats["avgExtF1Bases"]))
#write seed
if len(bestF1E.seq) < 50:
logging.warning("f1e sequence is small (%dbp) can't call consensus" % (len(bestF1E.seq)))
#I don't know what to return here
f1sfout = NamedTemporaryFile(prefix="flank1_", suffix=".fasta", delete=False, dir=basedir)
ALLTEMPFILES.append(f1sfout.name)
f1sfout.write(">%s\n%s\n" % (bestF1E.name, bestF1E.seq))
f1sfout.close()
f1sfout = f1sfout.name
#if stats["extendF2Count"] > 0:
if bestF2E is not None:
stats["avgExtF2Bases"] = stats["avgExtF2Bases"]/stats["extendF2Count"]
logging.info("estimated flank 2 extend len %d" % (stats["avgExtF2Bases"]))
#write seed
if len(bestF2E.seq) < 50:
logging.warning("f2e sequence is small (%dbp) can't call consensus" % (len(bestF2E.seq)))
#I don't know what to return here
f2sfout = NamedTemporaryFile(prefix="flank2", suffix=".fasta", delete=False, dir=basedir)
ALLTEMPFILES.append(f2sfout.name)
f2sfout.write(">%s\n%s\n" % (bestF2E.name, bestF2E.seq))
f2sfout.close()
f2sfout = f2sfout.name
#all of the info I need to return... refactor later and create useful objects
#ret = nt(stats, sfout, f1fout, f2fout, ssfout, f1sfout, f2sfout)
ret = nt(stats, ssfout, f1sfout, f2sfout)
#seeds writing
return ret
def buildFillSeq(data, inputReads, args):
"""
Using all of the information in the namedtuple returned from getSubSeqs,
go through the process of building the filling sequence.
load the filling sequence in to the data
"""
#try to build span
if SUPPORTFLAGS.span in data.stats["support"][0]:
logging.debug("build span")
alignFile = NamedTemporaryFile(prefix="scon_", suffix=".m5", delete=False, dir=args.tempDir)
alignFile.close(); alignFile = alignFile.name
ALLTEMPFILES.append(alignFile)
#blasr(data.spanReads, data.spanSeed, bestn = 1, nproc = args.nproc, outname=alignFile)
blasr(inputReads, data.spanSeed, bestn = 1, nproc = args.nproc, outname=alignFile)
aligns = M5File(alignFile)
if len(aligns) > 0:
con = consensus(aligns)
#if successful we're done
if con.contribBases > 0 and con.fillBases > 0:#must be
sequence = con.sequence#strandCorrector(data.stats["spanSeedStrand1"], con.sequence)
data.stats["fillSeq"] = sequence
data.stats["contribSeqs"] = con.contribSeqs
data.stats["contribBases"] = con.contribBases
data.stats["fillBases"] = con.fillBases
return
else:
logging.info("no mapping... picking span seq")
sequence = FastaFile(data.spanSeed).values()[0]
data.stats["fillSeq"] = sequence
data.stats["contribSeqs"] = 1
data.stats["contribBases"] = len(sequence)
data.stats["fillBases"] = len(sequence)
return
#no span -- we need to do flanks
flank1Success = False
flank2Success = False
logging.debug(json.dumps(data.stats, indent=4))
fl1Flag = SUPPORTFLAGS.left if data.stats["seed1"].endswith("e5") else SUPPORTFLAGS.right
if data.stats["seed2"] is not None:
fl2Flag = SUPPORTFLAGS.left if data.stats["seed2"].endswith("e5") else SUPPORTFLAGS.right
else:
fl2Flag = None
logging.debug((fl1Flag, fl2Flag))
if fl1Flag in data.stats["support"][1]:
logging.debug("build flank1 %d" % fl1Flag)
alignFile = NamedTemporaryFile(prefix="f1con_", suffix=".m5", delete=False, dir=args.tempDir)
alignFile.close(); alignFile = alignFile.name
ALLTEMPFILES.append(alignFile)
#blasr(data.flank1Reads, data.flank1Seed, bestn=1, nproc=args.nproc, outname=alignFile)
blasr(inputReads, data.flank1Seed, bestn=1, nproc=args.nproc, outname=alignFile)
aligns = M5File(alignFile)
if len(aligns) > 0:
con = consensus(aligns)
if con.contribBases > 0 and con.fillBases > 0:#must be
sequence = con.sequence#strandCorrector(data.stats["extendF1SeedStrand"], con.sequence)
data.stats["extendSeq1"] = sequence
data.stats["contribSeqs"] += con.contribSeqs
data.stats["contribBases"] += con.contribBases
data.stats["fillBases"] += con.fillBases
flank1Success = True
else:
logging.info("no mapping... picking f1 seq")
sequence = FastaFile(data.flank1Seed).values()[0]
data.stats["extendSeq1"] = sequence
data.stats["contribSeqs"] = 1
data.stats["contribBases"] = len(sequence)
data.stats["fillBases"] = len(sequence)
flank1Success = True
if fl2Flag in data.stats["support"][2]:
logging.debug("build flank2 %d" % fl2Flag)
alignFile = NamedTemporaryFile(prefix="f2con_", suffix=".m5", delete=False, dir=args.tempDir)
alignFile.close(); alignFile = alignFile.name
ALLTEMPFILES.append(alignFile)
#blasr(data.flank2Reads, data.flank2Seed, bestn=1, nproc=args.nproc, outname=alignFile)
blasr(inputReads, data.flank2Seed, bestn=1, nproc=args.nproc, outname=alignFile)
aligns = M5File(alignFile)
if len(aligns) > 0:
con = consensus(aligns)
if con.contribBases > 0 and con.fillBases > 0:#must be
sequence = con.sequence#strandCorrector(data.stats["extendF2SeedStrand"], con.sequence)
data.stats["extendSeq2"] = sequence
data.stats["contribSeqs"] += con.contribSeqs
data.stats["contribBases"] += con.contribBases
data.stats["fillBases"] += con.fillBases
flank2Success = True
else:
logging.info("no mapping... picking f1 seq")
sequence = FastaFile(data.flank2Seed).values()[0]
data.stats["extendSeq2"] = sequence
data.stats["contribSeqs"] = 1
data.stats["contribBases"] = len(sequence)
data.stats["fillBases"] = len(sequence)
flank2Success = True
if flank1Success and flank2Success:
logging.debug("mid unite")
seq = singleOverlapAssembly(data, args)
if seq is not None:
data.stats["fillSeq"] = seq
return
def strandCorrector(strand, sequence):
"""
ensures that the sequence inside of data is from the same strand as the
first seed
if -, flip it
"""
logging.debug("Weird %s" % (strand))
if strand == '1':
sequence = sequence.translate(revComp)[::-1]
return sequence
def singleOverlapAssembly(alldata, args):
"""
"""
global ALLTEMPFILES
data = alldata.stats
reads = NamedTemporaryFile(prefix="sol_", suffix=".fasta", delete=False, dir=args.tempDir)
ALLTEMPFILES.append(reads.name)
e1Seq = data["extendSeq1"]; e2Seq = data["extendSeq2"]
reads.write(">%s\n%s\n>%s\n%s\n" % ("seq1", e1Seq, "seq2", e2Seq))
reads.close()
alignFn = NamedTemporaryFile(prefix="sol_",suffix=".m5", delete=False, dir=args.tempDir)
ALLTEMPFILES.append(alignFn.name)
blasr(reads.name, reads.name, nproc=args.nproc, outname=alignFn.name)
aligns = M5File(alignFn)
# find best hit between the two
connector = AlignmentConnector()
bestS = None
bestA = 0
for i in aligns:
if i.qname != i.tname:
if connector.extendsTarget(i):
if i.score < bestS:
bestA = i
bestS = i.score
if bestS is None:
logging.info("no overlap between extenders")
return
#any of these steps could fail --
#Ensure the hit is valid
#(if + + and sameStrand we are okay, if - + and not sameStrand we are okay)
if data["sameStrand"] == (bestA.tstrand == '0'):
logging.info("bad overlap between extenders")
return
con = consensus([bestA])
bestA = bestA[0]
#strand correction...
if bestA.qname == "seq1":
if bestA.tstrand == '1':
e2Seq = e2Seq[:bestA.tstart].translate(revComp)[::-1]
seq = e1Seq[:bestA.qstart] + con.sequence.translate(revComp)[::-1] + e2Seq
else:
seq = e1Seq[:bestA.qstart] + con.sequence + e2Seq[bestA.tend:]
else:
if bestA.tstrand == '1':
e2Seq = e2Seq[:bestA.qstart].translate(revComp)[::-1]
seq = e1Seq[:bestA.tstart] + con.sequence + e2Seq
else:
seq = e1Seq[:bestA.qstart] + con.sequence + e2Seq[bestA.tstart:]
return seq
def preunitereads(inputFastq, args):
"""
sent query, I'm going to pop all of the united reads onto this
"""
global ALLTEMPFILES
alignFile = NamedTemporaryFile(prefix="uni_", suffix=".m5", delete=False, dir=args.tempDir).name
ALLTEMPFILES.append(alignFile)
readFile = NamedTemporaryFile(prefix="uni_", suffix=".fasta", delete=False, dir=args.tempDir)
ALLTEMPFILES.append(readFile.name)
input = FastqFile(inputFastq)
for read in input:
readFile.write(">%s\n%s\n" % (input[read].name, input[read].seq))
readFile.close()
readFile = readFile.name
blasr(readFile, readFile, bestn=5, nCandidates=20, nproc=args.nproc, outname=alignFile)
aligns = M5File(alignFile)
con = AlignmentConnector()
extenders = []
for a in aligns:
if a.tname == a.qname:
continue
if a.qstart - a.qend < 500 or a.tstart - a.tend < 500:
continue
sup = con.extendsTarget(a, minCovers=500)
#sup = con.extendsTarget(a, minCovers=100)
a.support = sup
if sup in [SUPPORTFLAGS.left, SUPPORTFLAGS.right]:
extenders.append(a)
best = {}#best of queries
for i in extenders:
score = 0
if i.qname in best:
score = best[i.qname].score
if i.score < score:
best[i.qname] = i
#print "q"
#for i in best.values():
#print str(i)
best2 = {}#best of targets
for i in best.values():
score = 0
if i.tname in best2:
score = best2[i.tname].score
if i.score < score:
best2[i.tname] = i
#print "t"
#for i in best2.values():
#print str(i)
best3 = {}#best of both
for i in best2.values():
keys = [i.qname, i.tname]
keys.sort()
keys = "".join(keys)
score = 0
if keys in best3:
score = best3[keys].score
if i.score < score:
best3[keys] = i
#print 'b'
#for i in best3.values():
#print str(i)
reads = FastqFile(inputFastq)
fout = open(inputFastq, 'a')
count = 0
for i in best3.values():
qseq = None
if i.support == SUPPORTFLAGS.left:
if i.qstrand == '0':
qseq = reads[i.qname].seq + reads[i.tname].seq[i.tend:]
elif i.qstrand == '1':
qseq = reads[i.qname].seq + reads[i.tname].seq[i.tend:].translate(revComp)
if i.support == SUPPORTFLAGS.right:
if i.qstrand == '0':
qseq = reads[i.tname].seq[:i.tstart] + reads[i.qname].seq
elif i.qstrand == '1':
qseq = reads[i.tname].seq[:i.tstart].translate(revComp) + reads[i.qname].seq
if qseq is not None:
count += 1
fout.write("@%s_%s\n%s\n+\n%s\n" % (i.qname, i.tname, qseq, "!"*len(qseq)))
logging.info("Preunited %d reads" % (count))
fout.close()
def parseArgs():
"""
input dir
predicted gapsize
if argument says that we need to extract the seeds we will have a single paramters
extractFlanks
"""
parser = argparse.ArgumentParser(description=USAGE, \
formatter_class=argparse.RawDescriptionHelpFormatter)
parser.add_argument("asmdir", metavar="DIR", type=str, \
help="Local assembly directory for a gap")
parser.add_argument("-t", "--maxTrim", type=int, default=100, \
help="Maxmum trim allowed (100)")
parser.add_argument("-w", "--maxWiggle", type=int, default=400, \
help="Maxmum wiggle for gap spanning allowed (400)")
parser.add_argument("-p", "--predictedGapSize", type=int, default=None)
parser.add_argument("-n", "--nproc", type=int, default=1)
parser.add_argument("-k", "--keepTemp", action="store_true",\
help="Keep temporary files")
parser.add_argument("--tempDir", type=str, default=None,
help="Where to write temporary files (DIR)")
parser.add_argument("--debug", action="store_true")
args = parser.parse_args()
if args.asmdir.endswith("/"):
args.asmdir = args.asmdir[:-1]
if args.tempDir is None:
args.tempDir = args.asmdir
setupLogging(args.debug)
return args
def run():
global ALLTEMPFILES
args = parseArgs()
dirName = os.path.basename(args.asmdir)
sameStrand, seeds = orderSeeds(dirName.split('_'))
inputReads = FastqFile(os.path.join(args.asmdir,"input.fastq"))
supportFn, flankFn = extractFlanks(inputReads, basedir=args.tempDir)
preunitereads(supportFn, args)
onFlank = NamedTemporaryFile(prefix="onFlank_", suffix=".m5", delete=False, dir=args.tempDir)
ALLTEMPFILES.append(onFlank.name)
onFlank.close()
tailblasr(supportFn, flankFn, nproc=args.nproc, \
outname=onFlank.name, basedir=args.tempDir)
data = getSubSeqs(onFlank.name, supportFn, sameStrand, seeds, \
args.predictedGapSize, args.maxTrim, args.maxWiggle, basedir=args.tempDir)
if data.stats["spanSeedName"] != "tooShortNs":
buildFillSeq(data, supportFn, args)
#if data.stats["support"][0] == SUPPORTFLAGS.span:
#logging.info("spanned gap")
#else:
#logging.info("seed1 extend %d - seed2 extend %d" % tuple(data.stats["support"][1:]))
data.stats["predictedGapSize"] = args.predictedGapSize
jOut = open(os.path.join(args.asmdir, "fillingMetrics.json"),'w')
jOut.write(json.dumps(data.stats,indent=4))
jOut.close()
if not args.keepTemp:
logging.info("Cleaning %d temp files" % (len(ALLTEMPFILES)))
for i in ALLTEMPFILES:
os.remove(i)
logging.info("Finished")
if __name__ == '__main__':
run()
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