File: adapters.py

package info (click to toggle)
python-cutadapt 1.12-2~bpo8%2B1
  • links: PTS, VCS
  • area: main
  • in suites: jessie-backports
  • size: 2,112 kB
  • sloc: python: 4,297; makefile: 166
file content (569 lines) | stat: -rw-r--r-- 19,387 bytes parent folder | download | duplicates (2)
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
# coding: utf-8
"""
Adapters
"""
from __future__ import print_function, division, absolute_import
import sys
import re
from collections import defaultdict
from cutadapt import align, colorspace
from cutadapt.seqio import ColorspaceSequence, FastaReader

# Constants for the find_best_alignment function.
# The function is called with SEQ1 as the adapter, SEQ2 as the read.
# TODO get rid of those constants, use strings instead
BACK = align.START_WITHIN_SEQ2 | align.STOP_WITHIN_SEQ2 | align.STOP_WITHIN_SEQ1
FRONT = align.START_WITHIN_SEQ2 | align.STOP_WITHIN_SEQ2 | align.START_WITHIN_SEQ1
PREFIX = align.STOP_WITHIN_SEQ2
SUFFIX = align.START_WITHIN_SEQ2
ANYWHERE = align.SEMIGLOBAL
LINKED = 'linked'


def parse_braces(sequence):
	"""
	Replace all occurrences of ``x{n}`` (where x is any character) with n
	occurrences of x. Raise ValueError if the expression cannot be parsed.

	>>> parse_braces('TGA{5}CT')
	TGAAAAACT
	"""
	# Simple DFA with four states, encoded in prev
	result = ''
	prev = None
	for s in re.split('(\{|\})', sequence):
		if s == '':
			continue
		if prev is None:
			if s == '{':
				raise ValueError('"{" must be used after a character')
			if s == '}':
				raise ValueError('"}" cannot be used here')
			prev = s
			result += s
		elif prev == '{':
			prev = int(s)
			if not 0 <= prev <= 10000:
				raise ValueError('Value {} invalid'.format(prev))
		elif isinstance(prev, int):
			if s != '}':
				raise ValueError('"}" expected')
			result = result[:-1] + result[-1] * prev
			prev = None
		else:
			if s != '{':
				raise ValueError('Expected "{"')
			prev = '{'
	# Check if we are in a non-terminating state
	if isinstance(prev, int) or prev == '{':
		raise ValueError("Unterminated expression")
	return result


class AdapterParser(object):
	"""
	Factory for Adapter classes that all use the same parameters (error rate,
	indels etc.). The given **kwargs will be passed to the Adapter constructors.
	"""
	def __init__(self, colorspace=False, **kwargs):
		self.colorspace = colorspace
		self.constructor_args = kwargs
		self.adapter_class = ColorspaceAdapter if colorspace else Adapter

	def _parse_no_file(self, spec, name=None, cmdline_type='back'):
		"""
		Parse an adapter specification not using ``file:`` notation and return
		an object of an appropriate Adapter class. The notation for anchored
		5' and 3' adapters is supported. If the name parameter is None, then
		an attempt is made to extract the name from the specification
		(If spec is 'name=ADAPTER', name will be 'name'.)

		cmdline_type -- describes which commandline parameter was used (``-a``
		is 'back', ``-b`` is 'anywhere', and ``-g`` is 'front').
		"""
		if name is None:
			name, spec = self._extract_name(spec)
		sequence = spec
		types = dict(back=BACK, front=FRONT, anywhere=ANYWHERE)
		if cmdline_type not in types:
			raise ValueError('cmdline_type cannot be {0!r}'.format(cmdline_type))
		where = types[cmdline_type]
		if where == FRONT and spec.startswith('^'):  # -g ^ADAPTER
			sequence, where = spec[1:], PREFIX
		elif where == BACK:
			sequence1, middle, sequence2 = spec.partition('...')
			if middle == '...':
				if not sequence1:  # -a ...ADAPTER
					sequence = sequence1[3:]
				elif not sequence2:  # -a ADAPTER...
					sequence, where = spec[:-3], PREFIX
				else:  # -a ADAPTER1...ADAPTER2
					if self.colorspace:
						raise NotImplementedError('Using linked adapters in colorspace is not supported')
					if sequence1.startswith('^') or sequence2.endswith('$'):
						raise NotImplementedError('Using "$" or "^" when '
							'specifying a linked adapter is not supported')
					return LinkedAdapter(sequence1, sequence2, name=name,
						**self.constructor_args)
			elif spec.endswith('$'):   # -a ADAPTER$
				sequence, where = spec[:-1], SUFFIX
		if not sequence:
			raise ValueError("The adapter sequence is empty.")

		return self.adapter_class(sequence, where, name=name, **self.constructor_args)

	def parse(self, spec, cmdline_type='back'):
		"""
		Parse an adapter specification and yield appropriate Adapter classes.
		This works like the _parse_no_file() function above, but also supports the
		``file:`` notation for reading adapters from an external FASTA
		file. Since a file can contain multiple adapters, this
		function is a generator.
		"""
		if spec.startswith('file:'):
			# read adapter sequences from a file
			with FastaReader(spec[5:]) as fasta:
				for record in fasta:
					name = record.name.split(None, 1)[0]
					yield self._parse_no_file(record.sequence, name, cmdline_type)
		else:
			name, spec = self._extract_name(spec)
			yield self._parse_no_file(spec, name, cmdline_type)

	def _extract_name(self, spec):
		"""
		Parse an adapter specification given as 'name=adapt' into 'name' and 'adapt'.
		"""
		fields = spec.split('=', 1)
		if len(fields) > 1:
			name, spec = fields
			name = name.strip()
		else:
			name = None
		spec = spec.strip()
		return name, spec

	def parse_multi(self, back, anywhere, front):
		"""
		Parse all three types of commandline options that can be used to
		specify adapters. back, anywhere and front are lists of strings,
		corresponding to the respective commandline types (-a, -b, -g).

		Return a list of appropriate Adapter classes.
		"""
		adapters = []
		for specs, cmdline_type in (back, 'back'), (anywhere, 'anywhere'), (front, 'front'):
			for spec in specs:
				adapters.extend(self.parse(spec, cmdline_type))
		return adapters


class Match(object):
	"""
	TODO creating instances of this class is relatively slow and responsible for quite some runtime.
	"""
	__slots__ = ['astart', 'astop', 'rstart', 'rstop', 'matches', 'errors', 'front', 'adapter', 'read', 'length']
	def __init__(self, astart, astop, rstart, rstop, matches, errors, front, adapter, read):
		self.astart = astart
		self.astop = astop
		self.rstart = rstart
		self.rstop = rstop
		self.matches = matches
		self.errors = errors
		self.front = self._guess_is_front() if front is None else front
		self.adapter = adapter
		self.read = read
		# Number of aligned characters in the adapter. If there are
		# indels, this may be different from the number of characters
		# in the read.
		self.length = self.astop - self.astart
		assert self.length > 0
		assert self.errors / self.length <= self.adapter.max_error_rate
		assert self.length - self.errors > 0

	def __str__(self):
		return 'Match(astart={0}, astop={1}, rstart={2}, rstop={3}, matches={4}, errors={5})'.format(
			self.astart, self.astop, self.rstart, self.rstop, self.matches, self.errors)

	def _guess_is_front(self):
		"""
		Return whether this is guessed to be a front adapter.

		The match is assumed to be a front adapter when the first base of
		the read is involved in the alignment to the adapter.
		"""
		return self.rstart == 0

	def wildcards(self, wildcard_char='N'):
		"""
		Return a string that contains, for each wildcard character,
		the character that it matches. For example, if the adapter
		ATNGNA matches ATCGTA, then the string 'CT' is returned.

		If there are indels, this is not reliable as the full alignment
		is not available.
		"""
		wildcards = [ self.read.sequence[self.rstart + i:self.rstart + i + 1] for i in range(self.length)
			if self.adapter.sequence[self.astart + i] == wildcard_char and self.rstart + i < len(self.read.sequence) ]
		return ''.join(wildcards)

	def rest(self):
		"""
		Return the part of the read before this match if this is a
		'front' (5') adapter,
		return the part after the match if this is not a 'front' adapter (3').
		This can be an empty string.
		"""
		if self.front:
			return self.read.sequence[:self.rstart]
		else:
			return self.read.sequence[self.rstop:]
	
	def get_info_record(self):
		seq = self.read.sequence
		qualities = self.read.qualities
		info = (
			self.read.name,
			self.errors,
			self.rstart,
			self.rstop,
			seq[0:self.rstart],
			seq[self.rstart:self.rstop],
			seq[self.rstop:],
			self.adapter.name
		)
		if qualities:
			info += (
				qualities[0:self.rstart],
				qualities[self.rstart:self.rstop],
				qualities[self.rstop:]
			)
		else:
			info += ('','','')
		
		return info

def _generate_adapter_name(_start=[1]):
	name = str(_start[0])
	_start[0] += 1
	return name


class Adapter(object):
	"""
	An adapter knows how to match itself to a read.
	In particular, it knows where it should be within the read and how to interpret
	wildcard characters.

	where --  One of the BACK, FRONT, PREFIX, SUFFIX or ANYWHERE constants.
		This influences where the adapter is allowed to appear within in the
		read and also which part of the read is removed.

	sequence -- The adapter sequence as string. Will be converted to uppercase.
		Also, Us will be converted to Ts.

	max_error_rate -- Maximum allowed error rate. The error rate is
		the number of errors in the alignment divided by the length
		of the part of the alignment that matches the adapter.

	minimum_overlap -- Minimum length of the part of the alignment
		that matches the adapter.

	read_wildcards -- Whether IUPAC wildcards in the read are allowed.

	adapter_wildcards -- Whether IUPAC wildcards in the adapter are
		allowed.

	name -- optional name of the adapter. If not provided, the name is set to a
		unique number.
	"""
	def __init__(self, sequence, where, max_error_rate=0.1, min_overlap=3,
			read_wildcards=False, adapter_wildcards=True, name=None, indels=True):
		self.debug = False
		self.name = _generate_adapter_name() if name is None else name
		self.sequence = parse_braces(sequence.upper().replace('U', 'T'))
		assert len(self.sequence) > 0
		self.where = where
		self.max_error_rate = max_error_rate
		self.min_overlap = min(min_overlap, len(self.sequence))
		self.indels = indels
		self.adapter_wildcards = adapter_wildcards and not set(self.sequence) <= set('ACGT')
		self.read_wildcards = read_wildcards
		# redirect trimmed() to appropriate function depending on adapter type
		trimmers = {
			FRONT: self._trimmed_front,
			PREFIX: self._trimmed_front,
			BACK: self._trimmed_back,
			SUFFIX: self._trimmed_back,
			ANYWHERE: self._trimmed_anywhere
		}
		self.trimmed = trimmers[where]
		if where == ANYWHERE:
			self._front_flag = None  # means: guess
		else:
			self._front_flag = where not in (BACK, SUFFIX)
		# statistics about length of removed sequences
		self.lengths_front = defaultdict(int)
		self.lengths_back = defaultdict(int)
		self.errors_front = defaultdict(lambda: defaultdict(int))
		self.errors_back = defaultdict(lambda: defaultdict(int))
		self.adjacent_bases = { 'A': 0, 'C': 0, 'G': 0, 'T': 0, '': 0 }

		self.aligner = align.Aligner(self.sequence, self.max_error_rate,
			flags=self.where, wildcard_ref=self.adapter_wildcards, wildcard_query=self.read_wildcards)
		self.aligner.min_overlap = self.min_overlap
		if not self.indels:
			# TODO
			# When indels are disallowed, an entirely different algorithm
			# should be used.
			self.aligner.indel_cost = 100000

	def __repr__(self):
		return '<Adapter(name="{name}", sequence="{sequence}", where={where}, '\
			'max_error_rate={max_error_rate}, min_overlap={min_overlap}, '\
			'read_wildcards={read_wildcards}, '\
			'adapter_wildcards={adapter_wildcards}, '\
			'indels={indels})>'.format(**vars(self))

	def enable_debug(self):
		"""
		Print out the dynamic programming matrix after matching a read to an
		adapter.
		"""
		self.debug = True
		self.aligner.enable_debug()

	def match_to(self, read):
		"""
		Attempt to match this adapter to the given read.

		Return an Match instance if a match was found;
		return None if no match was found given the matching criteria (minimum
		overlap length, maximum error rate).
		"""
		read_seq = read.sequence.upper()
		pos = -1
		# try to find an exact match first unless wildcards are allowed
		if not self.adapter_wildcards:
			if self.where == PREFIX:
				pos = 0 if read_seq.startswith(self.sequence) else -1
			elif self.where == SUFFIX:
				pos = (len(read_seq) - len(self.sequence)) if read_seq.endswith(self.sequence) else -1
			else:
				pos = read_seq.find(self.sequence)
		if pos >= 0:
			match = Match(
				0, len(self.sequence), pos, pos + len(self.sequence),
				len(self.sequence), 0, self._front_flag, self, read)
		else:
			# try approximate matching
			if not self.indels and self.where in (PREFIX, SUFFIX):
				if self.where == PREFIX:
					alignment = align.compare_prefixes(self.sequence, read_seq,
						wildcard_ref=self.adapter_wildcards, wildcard_query=self.read_wildcards)
				else:
					alignment = align.compare_suffixes(self.sequence, read_seq,
						wildcard_ref=self.adapter_wildcards, wildcard_query=self.read_wildcards)
				astart, astop, rstart, rstop, matches, errors = alignment
				if astop - astart >= self.min_overlap and errors / (astop - astart) <= self.max_error_rate:
					match = Match(*(alignment + (self._front_flag, self, read)))
				else:
					match = None
			else:
				alignment = self.aligner.locate(read_seq)
				if self.debug:
					print(self.aligner.dpmatrix)  # pragma: no cover
				if alignment is None:
					match = None
				else:
					astart, astop, rstart, rstop, matches, errors = alignment
					match = Match(astart, astop, rstart, rstop, matches, errors, self._front_flag, self, read)

		if match is None:
			return None
		assert match.length > 0 and match.errors / match.length <= self.max_error_rate, match
		assert match.length >= self.min_overlap
		return match

	def _trimmed_anywhere(self, match):
		"""Return a trimmed read"""
		if match.front:
			return self._trimmed_front(match)
		else:
			return self._trimmed_back(match)

	def _trimmed_front(self, match):
		"""Return a trimmed read"""
		# TODO move away
		self.lengths_front[match.rstop] += 1
		self.errors_front[match.rstop][match.errors] += 1
		return match.read[match.rstop:]

	def _trimmed_back(self, match):
		"""Return a trimmed read without the 3' (back) adapter"""
		# TODO move away
		self.lengths_back[len(match.read) - match.rstart] += 1
		self.errors_back[len(match.read) - match.rstart][match.errors] += 1
		adjacent_base = match.read.sequence[match.rstart-1:match.rstart]
		if adjacent_base not in 'ACGT':
			adjacent_base = ''
		self.adjacent_bases[adjacent_base] += 1
		return match.read[:match.rstart]

	def __len__(self):
		return len(self.sequence)


class ColorspaceAdapter(Adapter):
	def __init__(self, *args, **kwargs):
		super(ColorspaceAdapter, self).__init__(*args, **kwargs)
		has_nucleotide_seq = False
		if set(self.sequence) <= set('ACGT'):
			# adapter was given in basespace
			self.nucleotide_sequence = self.sequence
			has_nucleotide_seq = True
			self.sequence = colorspace.encode(self.sequence)[1:]
		if self.where in (PREFIX, FRONT) and not has_nucleotide_seq:
			raise ValueError("A 5' colorspace adapter needs to be given in nucleotide space")
		self.aligner.reference = self.sequence

	def match_to(self, read):
		"""Return Match instance"""
		if self.where != PREFIX:
			return super(ColorspaceAdapter, self).match_to(read)
		# create artificial adapter that includes a first color that encodes the
		# transition from primer base into adapter
		asequence = colorspace.ENCODE[read.primer + self.nucleotide_sequence[0:1]] + self.sequence

		pos = 0 if read.sequence.startswith(asequence) else -1
		if pos >= 0:
			match = Match(
				0, len(asequence), pos, pos + len(asequence),
				len(asequence), 0, self._front_flag, self, read)
		else:
			# try approximate matching
			self.aligner.reference = asequence
			alignment = self.aligner.locate(read.sequence)
			if self.debug:
				print(self.aligner.dpmatrix)  # pragma: no cover
			if alignment is not None:
				match = Match(*(alignment + (self._front_flag, self, read)))
			else:
				match = None

		if match is None:
			return None
		assert match.length > 0 and match.errors / match.length <= self.max_error_rate
		assert match.length >= self.min_overlap
		return match

	def _trimmed_front(self, match):
		"""Return a trimmed read"""
		read = match.read
		self.lengths_front[match.rstop] += 1
		self.errors_front[match.rstop][match.errors] += 1
		# to remove a front adapter, we need to re-encode the first color following the adapter match
		color_after_adapter = read.sequence[match.rstop:match.rstop + 1]
		if not color_after_adapter:
			# the read is empty
			return read[match.rstop:]
		base_after_adapter = colorspace.DECODE[self.nucleotide_sequence[-1:] + color_after_adapter]
		new_first_color = colorspace.ENCODE[read.primer + base_after_adapter]
		new_read = read[:]
		new_read.sequence = new_first_color + read.sequence[(match.rstop + 1):]
		new_read.qualities = read.qualities[match.rstop:] if read.qualities else None
		return new_read

	def _trimmed_back(self, match):
		"""Return a trimmed read"""
		# trim one more color if long enough
		adjusted_rstart = max(match.rstart - 1, 0)
		self.lengths_back[len(match.read) - adjusted_rstart] += 1
		self.errors_back[len(match.read) - adjusted_rstart][match.errors] += 1
		return match.read[:adjusted_rstart]

	def __repr__(self):
		return '<ColorspaceAdapter(sequence={0!r}, where={1})>'.format(self.sequence, self.where)


class LinkedMatch(object):
	"""
	Represent a match of a LinkedAdapter.

	TODO
	It shouldn’t be necessary to have both a Match and a LinkedMatch class.
	"""
	def __init__(self, front_match, back_match, adapter):
		self.front_match = front_match
		self.back_match = back_match
		self.adapter = adapter
		assert front_match is not None


class LinkedAdapter(object):
	"""
	"""
	def __init__(self, front_sequence, back_sequence,
			front_anchored=True, back_anchored=False, name=None, **kwargs):
		"""
		kwargs are passed on to individual Adapter constructors
		"""
		assert front_anchored and not back_anchored
		where1 = PREFIX if front_anchored else FRONT
		where2 = SUFFIX if back_anchored else BACK
		self.front_anchored = front_anchored
		self.back_anchored = back_anchored

		# The following attributes are needed for the report
		self.where = LINKED
		self.name = _generate_adapter_name() if name is None else name
		self.front_adapter = Adapter(front_sequence, where=where1, name=None, **kwargs)
		self.back_adapter = Adapter(back_sequence, where=where2, name=None, **kwargs)

	def enable_debug(self):
		self.front_adapter.enable_debug()
		self.back_adapter.enable_debug()

	def match_to(self, read):
		"""
		Match the linked adapters against the given read. If the 'front' adapter
		is not found, the 'back' adapter is not searched for.
		"""
		front_match = self.front_adapter.match_to(read)
		if front_match is None:
			return None
		# TODO use match.trimmed() instead as soon as that does not update
		# statistics anymore
		read = read[front_match.rstop:]
		back_match = self.back_adapter.match_to(read)
		return LinkedMatch(front_match, back_match, self)

	def trimmed(self, match):
		front_trimmed = self.front_adapter.trimmed(match.front_match)
		if match.back_match:
			return self.back_adapter.trimmed(match.back_match)
		else:
			return front_trimmed

	# Lots of forwarders (needed for the report). I’m sure this can be done
	# in a better way.

	@property
	def lengths_front(self):
		return self.front_adapter.lengths_front

	@property
	def lengths_back(self):
		return self.back_adapter.lengths_back

	@property
	def errors_front(self):
		return self.front_adapter.errors_front

	@property
	def errors_back(self):
		return self.back_adapter.errors_back

	@property
	def adjacent_bases(self):
		return self.back_adapter.adjacent_bases