1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538 539 540 541 542 543 544 545 546 547 548 549 550 551 552 553 554 555 556 557 558 559 560 561 562 563 564 565 566 567 568 569 570 571 572 573 574 575 576 577 578 579 580 581 582 583 584 585 586 587 588 589 590 591 592 593 594 595 596 597 598 599 600 601 602 603 604 605 606 607 608 609 610 611 612 613 614 615 616 617 618 619 620 621 622 623 624 625 626 627 628 629 630 631 632 633 634 635 636 637 638 639 640 641 642 643 644 645
|
#!/usr/bin/env python
import sys, os
import numpy as np
import argparse
import bisect
from collections import defaultdict
import re
SEED_LEN = 22
MIN_CLUST = 25
SAMPLE_RATE = 4000
CHUNK_LEN = 4000
#Guppy basecalled event stride and kmer length
BCE_STRIDE = 5
BCE_K = 4
MAX_CHUNK_DEF = 3
#Cigar parsing
CIG_OPS_STR = "MIDNSHP=X"
CIG_RE = re.compile("(\d+)(["+CIG_OPS_STR+"])")
CIG_OPS = set(CIG_OPS_STR)
CIG_INCR_ALL = {'M','=', 'X'}
CIG_INCR_RD = CIG_INCR_ALL | {'I','S'}
CIG_INCR_RF = CIG_INCR_ALL | {'D','N'}
class DebugParser:
CONF_PAD_COEF = 2
def __init__(self,
debug_prefix,
unc_paf,
mm2_paf=None,
bce_moves=None,
bwa_index=None,
min_chunk=None,
max_chunk=None,
min_samp=None,
max_samp=None,
tgt_cid=None,
max_path_fm=0,
load_seeds=True,
load_events=True,
load_paths=True,
load_conf=True):
self.rid = unc_paf.qr_name
#Cofident seed cluster ID, confident event, and reference length
#(conf_evt is where mapping would normally end)
self.conf_cid = unc_paf.tags.get('sc', (None,)*2)[0]
self.conf_evt = unc_paf.tags.get('ce', (None,)*2)[0]
self.conf_samp = None
self.mm2_paf = mm2_paf
self.idx = bwa_index
#if self.tgt_cid is None:
# self.tgt_cid = self.conf_cid
if min_samp is None and min_chunk is None:
self.min_samp = self.min_chunk = 0
elif min_chunk is None:
self.min_samp = min_samp
self.min_chunk = (min_samp-1) // CHUNK_LEN
elif min_samp is None:
self.min_samp = self.min_chunk * SAMPLE_RATE
self.min_chunk = min_chunk
else:
self.min_samp = max(min_samp, min_chunk * SAMPLE_RATE)
self.min_chunk = max(min_chunk, (min_samp-1) // CHUNK_LEN)
if max_samp is None and max_chunk is None:
if self.conf_evt is None:
self.max_chunk = MAX_CHUNK_DEF
self.max_samp = self.max_chunk*SAMPLE_RATE
else:
self.max_chunk = self.max_samp = None
elif max_samp is None:
self.max_chunk = max_chunk
self.max_samp = max_chunk * SAMPLE_RATE
elif max_chunk is None:
self.max_chunk = max(0, (max_samp-1) // CHUNK_LEN)
self.max_samp = max_samp
else:
self.max_chunk = max(max_chunk, (max_samp-1) // CHUNK_LEN)
self.max_samp = max(max_samp, self.max_chunk * SAMPLE_RATE)
self.min_evt = 0 if self.min_samp == 0 else None
self.max_evt = None
self.chunk_evt_bounds = [0]
#TODO below should be w/r/t reference coordinates?
#allow argument specified
#or detect based on confident cluster (or just specify seed cluster)
#TBD in parse_seeds
self.conf_len = None
self.conf_clust = None
self.max_clust = None
self.max_clust_len = None
if mm2_paf is not None:
self.ref_name = mm2_paf.rf_name
if mm2_paf.is_fwd:
self.min_ref = mm2_paf.rf_st
else:
self.min_ref = -mm2_paf.rf_en
else:
self.min_ref = None
self.max_ref = None
#TODO clean these up
self.conf_pbs = dict()
self.conf_dots = set()
self.path_lens = defaultdict(int)
self.seed_kmers = dict()
self.conf_ref_bounds = None
self.evts_loaded = False
if load_events:
self.evts_in = open(debug_prefix + self.rid + "_events.tsv")
self.parse_events()
self.seeds_loaded = False
if load_seeds:
self.seeds_in = open(debug_prefix + self.rid + "_seeds.bed")
self.parse_seeds()
self.bc_loaded = False
if mm2_paf is not None and bce_moves is not None:
self.parse_bc_aln(bce_moves)
self.max_path_fm = max_path_fm
if max_path_fm > 0:
if self.idx is None:
sys.stderr.write("Error: must include BWA index to include paths with FM overlaps\n")
sys.exit(1)
if self.conf_clust.fwd:
st = self.min_ref
en = self.max_ref
else:
st = -self.max_ref
en = -self.min_ref
fwd_fms, rev_fms = self.idx.range_to_fms(
self.ref_name, st, en
)
fms = fwd_fms if self.mm2_paf.is_fwd else rev_fms
self.fm_to_ref = {fms[i] : i for i in range(len(fms))}
self.range_fms = np.sort(fms)
else:
self.fwd_fms = self.rev_fms = None
#TODO: paths
self.paths_loaded = False
if load_paths:
self.paths_fname = debug_prefix + self.rid + "_paths.tsv"
self.parse_paths()
self.conf_loaded = False
if load_conf:
self.conf_fname = debug_prefix + self.rid + "_conf.tsv"
self.parse_conf()
def parse_events(self, incl_norm=True):
if self.evts_loaded: return False
self.evts_in.readline()
self.events = list()
self.norms = list()
self.win_means = list()
self.win_stdvs = list()
self.evt_mask = list()
self.evt_mask_map = list()
unmask_evts = 0
evt = 0
chunk = 0
next_chunk_samp = CHUNK_LEN
for line in self.evts_in:
tabs = line.split()
st,ln,mask = map(int, tabs[:2] + tabs[-1:])
mn,sd,norm_sc,norm_sh,win_mn,win_sd = map(float, tabs[2:-1])
en = st+ln
if st >= next_chunk_samp:
chunk += 1
next_chunk_samp += CHUNK_LEN
self.chunk_evt_bounds.append(evt)
if evt == self.conf_evt:
self.conf_samp = st
if self.max_samp is None:
self.max_samp = (chunk+1)*CHUNK_LEN-1
self.max_chunk = chunk
if en < self.min_samp:
evt += 1
continue
if self.min_evt is None:
print(en, self.min_samp)
self.min_evt = evt
print(self.min_evt, "MIN")
if self.max_samp is not None and st > self.max_samp:
break
self.events.append( (st,ln,mn,sd) )
self.norms.append( (norm_sc,norm_sh) )
self.win_means.append(win_mn)
self.win_stdvs.append(win_sd)
self.evt_mask.append(mask == 1)
if self.evt_mask[-1]:
self.evt_mask_map.append(evt)
evt += 1
if self.max_evt is None or self.max_evt >= evt:
self.max_evt = evt
if self.max_samp is None:
self.max_samp = st+ln
self.max_chunk = (self.max_samp-1) // CHUNK_LEN
self.evts_loaded = True
self.win_means = np.array(self.win_means)
self.win_stdvs = np.array(self.win_stdvs)
self.evt_mask = np.array(self.evt_mask)
self.evt_mask_map = np.array(self.evt_mask_map)
return True
def normed_event(self, e):
scale,shift = self.norms[e]
return scale*self.events[e][2]+shift
def parse_seeds(self, expire_coef=None):
if self.seeds_loaded: return False
if expire_coef != None:
SeedCluster.EXPIRE_COEF = expire_coef
self.clusts = dict()
self.clusts_exp = list()
clust_ids = set()
conf_clust = None
for line in self.seeds_in:
rf,st,en,name,strand = line.split()
st,en,evt,pb,cid = map(int, [st,en]+name.split(":"))
#handle expiration
#should this (still) be here?
clust = self.clusts.get(cid, None)
replace = (
clust is not None and
clust.expired(evt) and
(cid != self.conf_cid or evt < self.conf_evt)
)
if clust == None or replace:
if replace:
self.clusts_exp.append(clust)
clust = SeedCluster(evt,rf,st,en,strand=="+",cid)
self.clusts[cid] = clust
clust_ids.add(cid)
else:
clust.add_seed(evt,rf,st,en)
if clust.id == self.conf_cid:
conf_clust = clust
if clust.fwd:
#self.conf_pbs[(evt,pb)] = en
self.conf_pbs[(evt,pb)] = st
else:
#self.conf_pbs[(evt,pb)] = -st
self.conf_pbs[(evt,pb)] = -en + 1
if clust > self.max_clust:
self.max_clust = clust
self.mapped = conf_clust is not None
#Handle unmapped reads
#TODO: mark them more explicitly
if not self.mapped:
if self.max_clust is not None:
conf_clust = self.max_clust
self.conf_evt = conf_clust.evts[-1]
else:
self.conf_evt = self.max_evt
self._set_conf_clust(conf_clust)
self._parse_expired(clust_ids)
self.seeds_loaded = True
return True
#sets confident cluster related vars
def _set_conf_clust(self, cc):
self.conf_clust = cc
if cc is None: return
self.conf_idx = np.searchsorted(cc.evts, self.conf_evt, side='right')-1
self.conf_len = cc.lens[self.conf_idx]
#TODO use cc.evrf_ratio(e,r)
evt_span = self.conf_evt - cc.evt_st
if self.max_evt is None:
max_max_evt = self.conf_evt + (evt_span * self.CONF_PAD_COEF)
self.max_evt = int(np.round(min(cc.evt_en+1, max_max_evt)))
#TODO rename min/max_clust_ref
self.min_idx = np.searchsorted(cc.evts, self.min_evt, side='left')
self.max_idx = np.searchsorted(cc.evts, self.max_evt, side='right')-1
if cc.fwd:
rst = cc.rsts[self.min_idx] - SEED_LEN
ren = cc.rens[self.max_idx]+1
else:
rst = -cc.rsts[self.min_idx] - SEED_LEN
ren = -cc.rens[self.max_idx]-1
ref_span = ren-rst
if ref_span > 0:
self.evrf_ratio = (self.max_evt) / ref_span
else:
self.evrf_ratio = 1
self.max_clust_len = cc.lens[self.max_idx]
if self.min_ref is None:
self.min_ref = rst
if self.max_ref is None:
self.max_ref = ren
if self.ref_name is None:
self.ref_name = cc.ref
#finds clusters that should expire
#by the end of loaded events
def _parse_expired(self, clust_ids):
for cid in clust_ids:
c = self.clusts[cid]
if c.expired(self.max_evt):
self.clusts_exp.append(c)
del self.clusts[cid]
def _parse_moves(self, moves):
return np.flip()
def parse_paths(self):
if self.paths_loaded: return False
path_counts = list()
paths_fwd = open(self.paths_fname)
head_tabs = paths_fwd.readline().split()
C = {head_tabs[i] : i for i in range(len(head_tabs))}
#paths_fwd.close()
#paths_rev = FileReadBackwards(self.paths_fname)
#for line in paths_rev:
for line in paths_fwd:
tabs = line.split()
if tabs[0] == head_tabs[0]: continue
path_id = tuple(map(int, tabs[C['id']].split(':')))
ref_st = self.conf_pbs.get(path_id, None)
is_conf = ref_st is not None
if not is_conf and self.max_path_fm == 0: continue
evt, pb = path_id
moves = [bool(c=='1') for c in tabs[C['moves']]]
if evt < self.min_evt or evt >= self.max_evt:
continue
e = evt-self.min_evt
if not self.evt_mask[e-len(moves)]:
continue
evts = np.arange(e-len(moves), e) + 1
if not is_conf:
fm_len = int(tabs[C['fm_len']])
if fm_len > self.max_path_fm: continue
fm_start = int(tabs[C['fm_start']])
full_len = int(tabs[C['full_len']])
i = np.searchsorted(self.range_fms, fm_start)
while i < len(self.range_fms) and self.range_fms[i] < fm_start+fm_len:
ref_en = self.fm_to_ref[self.range_fms[i]]
refs = ref_en - np.sum(moves) + np.cumsum(np.flip(moves)) - 3
print(ref_en, path_id)
for e,r in zip(evts,refs):
if e < 0: continue
self.path_lens[(e,r)] = max(self.path_lens[(e,r)], np.log2(fm_len))
i += 1
else:
refs = ref_st + (np.cumsum(np.flip(moves)) - 1)
for e,r in zip(evts,refs):
if e < 0: continue
self.conf_dots.add( (e,r) )
#kmer = tabs[C['kmer']]
#match_prob = float(tabs[C['match_prob']])
#self.seed_kmers[(e, refs[-1])] = (kmer, match_prob)
#print(path_id,parent,fm_start,fm_len,kmer,full_len,seed_prob,moves)
# #Store number of paths at each event position
# while len(path_counts) - 1 < evt:
# path_counts.append(0)
# path_counts[-1] += 1
#self.paths_loaded = True
return True
def parse_conf(self):
if self.conf_loaded: return False
self.conf_evts = list()
self.conf_clusts = list()
self.conf_tops = list()
self.conf_means = list()
conf_in = open(self.conf_fname)
conf_in.readline()
for line in conf_in:
evt,clust,top,mean = line.split()
self.conf_evts.append(int(evt))
self.conf_clusts.append(int(clust))
self.conf_tops.append(float(top))
self.conf_means.append(float(mean))
def parse_bc_aln(self, bce_moves):
#List of ref coords for each query (read) cord
qr_to_rfs = self._cig_query_to_refs(self.mm2_paf)
#basecalled event start, end, and stride
bce_samp_st, bce_moves_pac = bce_moves
bce_samp_en = bce_samp_st + len(bce_moves) * BCE_STRIDE
bce_st = 0
bce_en = int((self.max_samp-bce_samp_st+1) // BCE_STRIDE)
bce_moves = np.unpackbits(bce_moves_pac)[bce_st:bce_en]
#Read coord of each basecalled event
bce_qrs = np.cumsum(bce_moves)
i = np.searchsorted(bce_qrs, self.mm2_paf.qr_st)
#bce_evts = list()
bce_samps = list()
bce_refs = list()
samp = bce_samp_st
for qr in bce_qrs:
if samp >= self.min_samp:
for rf in qr_to_rfs[qr]:
bce_samps.append(samp)
bce_refs.append(rf)
samp += BCE_STRIDE
self.bce_samps = np.array(bce_samps)
self.bce_refs = np.array(bce_refs) - BCE_K + 1
self.max_ref = self.min_ref + max(bce_refs)
def _cig_query_to_refs(self, paf):
cig = paf.tags.get('cg', (None,)*2)[0]
if cig is None: return None
qr_rfs = defaultdict(list)
qr_i = paf.qr_st
rf_i = 0#paf.rf_st
cig_ops = CIG_RE.findall(cig)
if not paf.is_fwd:
cig_ops = reversed(cig_ops)
for l,c in cig_ops:
l = int(l)
incr_qr = c in CIG_INCR_RD
incr_rf = c in CIG_INCR_RF
qr_j = qr_i + (l if incr_qr else 1)
rf_j = rf_i + (l if incr_rf else 1)
for qr,rf in zip(range(qr_i, qr_j), range(rf_i, rf_j)):
qr_rfs[qr].append(rf)
if incr_qr:
qr_i = qr_j
if incr_rf:
rf_i = rf_j
return qr_rfs
class SeedCluster:
EXPIRE_COEF = 5.5
#TODO add start evt at st-SEED_LEN?
#would make reference len easier to breakdown
#and could be nice for plotting?
def __init__(self, evt, rf, st, en, fwd, cid):
self.id = cid
self.rf = rf
self.evts = [evt]
self.blocks = [(st, en)] #TODO only use rsts?
self.gains = [en-st]
self.rsts = [st] #TODO consider strand
self.rens = [en] #TODO consider strand
self.lens = [self.gains[0]]
self.fwd = fwd
self.exp_evt = None
def expired(self, evt=None):
return False #
#if evt == None:
# return self.exp_evt is not None
#if evt - self.evt_en > self.ref_len * self.EXPIRE_COEF:
# if self.exp_evt is None:
# self.exp_evt = int(np.round(
# self.evt_en + (self.ref_len * self.EXPIRE_COEF)
# ))
# return True
#return False
def add_gain(self, gain):
self.gains.append(gain)
self.lens.append(len(self)+max(gain,0))
def add_seed(self, evt, rf, st, en):
bst, ben = self.blocks[-1]
self.evts.append(evt)
self.rsts.append(st)
self.rens.append(en)
if (self.rf != rf or max(bst, st) > min(ben, en)):
self.blocks.append( (st,en) )
self.add_gain(en-st)
else:
l1 = ben-bst
self.blocks[-1] = (min(st, bst), max(en, ben))
l2 = self.blocks[-1][1] - self.blocks[-1][0]
self.add_gain(l2-l1)
return True
#TODO add start evt at st-SEED_LEN?
@property
def ref_len(self):
return self.lens[-1]
@property
def evt_st(self):
return self.evts[0]-SEED_LEN
@property
def evt_en(self):
return self.evts[-1]
def evt_len(self, i=0, j=-1):
return self.evts[j] - self.evts[i] + 1
@property
def evrf_ratio(self, i=0, j=-1):
return (self.evts[j] - self.evt_st+1) / self.lens[i]
def __str__(self):
MAX_REF = 22
if len(self.rf) < MAX_REF:
ref = self.rf
else:
ref = self.rf[:MAX_REF] + "."
return "%s:%d-%d (%s)" % (
ref,
self.blocks[0][0],
self.blocks[-1][1],
"+" if self.fwd else "-")
#TODO bind confidence to len
#then make it more complicated
def __gt__(self, c):
return c is None or len(self) > len(c)
def __eq__(self, rhs):
return self.id == getattr(rhs, "id", None)
def __len__(self):
return self.lens[-1]
|