def test_unaligned_mafft_aa(self):
seq1 = SeqRecord(Seq('AC--DEFGI-LBATR'), id='1')
seq2 = SeqRecord(Seq('ACN-DEFGI--DATT'), id='2')
# 'AC-DEFGILBATR'
# 'ACNDEFGIDATT-'
e = {2: ('-', 'N'),
8: ('L', 'D'),
9: ('B', 'A'),
10: ('A', 'T'),
12: ('R', '-'),
}
diffs, l = seqstats.get_differences(seq1, seq2, aligned = False,
alphabet = alphabets.ProteinAlphabet(),
ignore_gaps = False,
aligner_tools = ['mafft'])
self.assertEqual(diffs, e)
self.assertEqual(l, 13)
diffs, l = seqstats.get_differences(seq1, seq2, aligned = False,
alphabet = alphabets.ProteinAlphabet(),
ignore_gaps = True,
aligner_tools = ['mafft'])
e = {8: ('L', 'D'),
9: ('B', 'A'),
10: ('A', 'T'),
}
self.assertEqual(diffs, e)
self.assertEqual(l, 11)
def test_aligned_aa(self):
seq1 = SeqRecord(Seq('AC--DEFGI-LBATR'), id='1')
seq2 = SeqRecord(Seq('ACN-DEFGI--DATT'), id='2')
e = {14:('R', 'T')}
diffs, l = seqstats.get_differences(seq1, seq2, aligned = True,
alphabet = alphabets.ProteinAlphabet())
self.assertEqual(diffs, e)
self.assertEqual(l, 11)
diffs, l = seqstats.get_differences(seq1, seq2, aligned = True,
alphabet = alphabets.ProteinAlphabet(),
ignore_gaps = False)
e = {14: ('R', 'T'),
2: ('-', 'N'),
10: ('L', '-')}
self.assertEqual(diffs, e)
self.assertEqual(l, 15)
def test_amino_acid_seqs(self):
seqs = [
SeqRecord(Seq('MILV*XQP*'), id='1'),
SeqRecord(Seq('MILV*XQQ*'), id='2'),
SeqRecord(Seq('MILV*XPP*'), id='3'),
]
expected = {}
expected['1'] = {'2': 1, '3': 1}
expected['2'] = {'1': 1, '3': 2}
expected['3'] = {'1': 1, '2': 2}
distance_iter = seqstats.pairwise_distance_iter(
seq_iter = seqs,
alphabet = alphabets.ProteinAlphabet(),
per_site = False,
aligned = True,
ignore_gaps = True)
for i, (seq1, seq2, d, drc) in enumerate(distance_iter):
self.assertEqual(drc, None)
self.assertEqual(
expected[seq1.id][seq2.id],
d)
self.assertEqual(i, 2)
def main_cli():
description = '{name} {version}\n\n{description}'.format(**_program_info)
parser = argparse.ArgumentParser(
description=description,
formatter_class=argparse.RawDescriptionHelpFormatter)
parser.add_argument('input_file',
metavar='INPUT-SEQ-FILE',
type=argparse_utils.arg_is_file,
help=('Input sequence file to be vetted.'))
comparison_args = parser.add_argument_group(
'Comparison Options',
'Options to control the number and nature of sequence comparisons')
comparison_args.add_argument(
'-n',
'--num-samples',
type=int,
default=0,
help=('The number of randomly sampled sequences to which each '
'sequence will be compared. If less than 1 (the defualt is '
'0), all pairwise comparisons will be performed. For very '
'large numbers of sequences, performing all pairwise '
'comparisons will take a long time. This option will speed '
'things up as long as the number specified is less than '
'about half of the number of input sequences. If the '
'number you are considering is close to half of the number '
'sequences, you should probably specify zero and do all '
'combinations. You should not specify a number greater than '
'half the number of sequences, because it will take longer '
'and be less thorough than the default.'))
comparison_args.add_argument(
'--seed',
action='store',
type=int,
help=('Random number seed to use for the analysis. This option '
'is only revelant if a number greater than 0 is specified '
'for the `-n/--num-samples` option.'))
comparison_args.add_argument(
'--compare-translated',
action='store_true',
help=('Compare amino acid sequences encoded by the longest '
'reading frame found in each sequence. To use this option, '
'`data-type` must be dna or rna. See "Translation Options" '
'for controlling how the longest reading frame of each '
'sequence is determined and translated.'))
comparison_args.add_argument('--check-ids',
action='store_true',
help=('Check sequence IDs for duplicates.'))
comparison_args.add_argument(
'--summarize-reading-frame-lengths',
action='store_true',
help=('Report the length of the longest reading frame of '
'each sequence. See "Translation Options" for controlling '
'how reading frames are determined.'))
comparison_args.add_argument(
'-g',
'--count-gaps',
action='store_true',
help=('Count gaps when calculating pairwise sequence distances. '
'The default is to calculate (number of differences '
'ignoring gaps / number of aligned sites ignoring sites '
'with gaps) for each pairwise comparison. When this option '
'is used, the distance is (number of differences including '
'gap differences / total number of aligned sites).'))
alignment_args = parser.add_argument_group(
'Alignment Options',
('These options control if/how sequences are to be aligned prior '
'to calculating distances.'))
alignment_args.add_argument(
'-a',
'--aligned',
action='store_true',
help=('Treat input sequences as aligned. I.e., do not perform '
'pairwise alignment before calculating distances between '
'sequences (except when calculating distances for reverse '
'and complemented sequences).'))
alignment_args.add_argument(
'--aligner',
type=argparse_utils.arg_is_executable,
help=('Path to alignment program executable to use for pairwise'
'alignments of sequences. '
'The default is to look for muscle and then mafft in PATH, '
'and if neither are found use the (slow) built-in '
'function. Even if the `-a`/`--aligned` option is '
'specified, the aligner will still be used for pairwise '
'alignments when calculating distances of reverse and '
'complemented sequences.'))
alignment_args.add_argument(
'--msa',
action='store_true',
help=('Perform a full multiple sequence alignemnt prior to '
'comparing sequences. The default is to align each '
'pair of sequences being compared. This option is '
'overruled by the `-a`/`--aligned` option. '
'If this option is used '
'the resulting alignment is written to file.'))
alignment_args.add_argument(
'--msa-aligner',
type=argparse_utils.arg_is_executable,
help=('Path to alignment program executable to use for full '
'multiple sequence alignment. '
'The default is to look for mafft and then muscle in PATH, '
'and if neither are found the program will exit with an '
'error message. If you do not have mafft or muscle '
'you cannot use this option. '
'This option is only used if the `-a`/`--aligned` option '
'is not specified, and the `--msa` option is specified.'))
translation_args = parser.add_argument_group(
'Translation Options',
('These options control translation from nucleotide to amino acid '
'sequences.'))
translation_args.add_argument(
'--table',
type=int,
choices=list(range(1, 7)) + list(range(9, 17)) + list(range(21, 26)),
default=1,
help=('The translation table to use for any options associated '
'with translating nucleotide sequences to amino acids. '
'Option should be the integer that corresponds to the '
'desired translation table according to NCBI '
'(http://www.ncbi.nlm.nih.gov/Taxonomy/Utils/wprintgc.cgi). '
'The default is 1 (the "standard" code).'))
translation_args.add_argument(
'--allow-partial',
action='store_true',
default=False,
help=('Allow partial reading frames at the beginning (no start '
'codon) and end (no stop codon) of sequences.'))
translation_args.add_argument(
'--read-after-stop',
action='store_true',
default=False,
help=('A new reading frame begins immediately after a stop codon. '
'The default is to start reading frame at next start codon '
'after a stop codon. This option might be useful for exons.'))
data_args = parser.add_argument_group(
'Data Options', ('Options specifying the input data type and format'))
data_args.add_argument(
'-d',
'--data-type',
type=str,
choices=VALID_DATA_TYPES,
default='dna',
help=('The type of sequence data. The default is dna. Valid '
'options include: {0}.'.format(', '.join(VALID_DATA_TYPES))))
data_args.add_argument(
'--format',
dest='input_format',
type=str,
choices=FILE_FORMATS.supported_formats,
help=('The format of the input sequence file. Valid options '
'include: {0}. By default, the format is guessed based on '
'the extension of the first input file. However, if '
'provided, this option will always take precedence over '
'the file extension.'.format(', '.join(
FILE_FORMATS.supported_formats))))
output_args = parser.add_argument_group(
'Output Options', 'Options for controlling output of program')
output_args.add_argument(
'-o',
'--output-dir',
type=argparse_utils.arg_is_dir,
help=('The directory in which all output files will be written. '
'The default is to use the directory of the input file.'))
messaging_args = parser.add_argument_group(
'Messaging Options', ('These options control verbosity of messaging.'))
messaging_args.add_argument(
'--log-frequency',
type=argparse_utils.arg_is_nonnegative_int,
default=1000,
help=('The frequency at which to log progress. Default is to log '
'every 1000 sequence comparisons.'))
messaging_args.add_argument('--quiet',
action='store_true',
help='Run without verbose messaging.')
messaging_args.add_argument('--debug',
action='store_true',
help='Run in debugging mode.')
args = parser.parse_args()
##########################################################################
## set up logging
from seqsift.utils.messaging import get_logger, LOGGING_LEVEL_ENV_VAR
os.environ[LOGGING_LEVEL_ENV_VAR] = "INFO"
if args.quiet:
os.environ[LOGGING_LEVEL_ENV_VAR] = "WARNING"
if args.debug:
os.environ[LOGGING_LEVEL_ENV_VAR] = "DEBUG"
log = get_logger(name=__name__)
##########################################################################
## package imports
from seqsift.utils import GLOBAL_RNG, dataio, functions, alphabets
from seqsift.seqops import seqsum, seqmod, seqstats
from seqsift.utils.fileio import OpenFile
##########################################################################
## handle args
## set seed if randomly sampling sequences
if args.num_samples > 0:
if not args.seed:
args.seed = random.randint(1, 999999999)
GLOBAL_RNG.seed(args.seed)
log.warning('Seed: {0}'.format(args.seed))
## get input file format
if not args.input_format:
args.input_format = FILE_FORMATS.get_format_from_file_object(
args.input_file)
if not args.input_format:
log.error("Could not determine input format.\n"
"You must either provide the input format\n"
"using the '--from' option or have a recognizable\n"
"file extension on the input file name.\n"
"Here are the supported file extensions:\n{0}".format(
str(FILE_FORMATS)))
sys.stderr.write(str(parser.print_help()))
sys.exit(1)
aligner_tools = ['muscle', 'mafft']
if args.aligner:
aligner_tools = [args.aligner]
full_aligner_tools = ['mafft', 'muscle']
if args.msa_aligner:
full_aligner_tools = [args.msa_aligner]
if not args.output_dir:
args.output_dir = os.path.dirname(args.input_file)
full_alignment_out_path = os.path.join(args.output_dir, 'seqvet-msa.txt')
alphabet = alphabets.DnaAlphabet()
if args.data_type in ['aa', 'protein']:
alphabet = alphabets.ProteinAlphabet()
if (args.summarize_reading_frame_lengths
and (not args.data_type in ['dna', 'rna'])):
log.error("`--summarize-reading-frame-lengths` is only compatible "
"with DNA or RNA.")
sys.stderr.write(str(parser.print_help()))
sys.exit(1)
if (args.compare_translated and (not args.data_type in ['dna', 'rna'])):
log.error("`-compare-translated` is only compatible with DNA or RNA.")
sys.stderr.write(str(parser.print_help()))
sys.exit(1)
##########################################################################
## heavy lifting
seqs = dataio.get_seq_iter([args.input_file],
format=args.input_format,
data_type=args.data_type)
if args.summarize_reading_frame_lengths:
log.info('Summarizing longest reading frame lengths...')
if not isinstance(seqs, dataio.BufferedIter):
seqs = dataio.BufferedIter(seqs)
lengths = seqsum.summarize_longest_read_lengths(
seqs,
table=args.table,
allow_partial=args.allow_partial,
require_start_after_stop=(not args.read_after_stop))
length_path = os.path.join(args.output_dir,
'seqvet-reading-frame-lengths.txt')
log.info('Writing longest reading frame lengths to file...')
with OpenFile(length_path, 'w') as out:
out.write('seq_id\tlrf\trev_comp_lrf\n')
for (l, rc_l, seq_id) in lengths:
out.write('{0}\t{1}\t{2}\n'.format(seq_id, l, rc_l))
if args.compare_translated:
log.info('Translating longest reading frames for distance '
'calculations...')
seqs = seqmod.translate_longest_reading_frames(
seqs,
table=args.table,
allow_partial=args.allow_partial,
require_start_after_stop=(not args.read_after_stop))
alphabet = alphabets.ProteinAlphabet()
if args.check_ids:
log.info('Checking sequence IDs...')
if not isinstance(seqs, dataio.BufferedIter):
seqs = dataio.BufferedIter(seqs)
dups = seqstats.get_duplicate_ids(seqs)
if len(dups) > 0:
dup_path = functions.get_new_path(
os.path.join(args.output_dir, 'seqvet-duplicate-ids.txt'))
log.warning('Duplicate IDs found! Writing them to '
'{0}'.format(dup_path))
with OpenFile(dup_path, 'w') as out:
for dup in dups:
out.write('{0}\n'.format(dup))
else:
log.info('No duplicate sequence IDs were found.')
log.info('Calculating pairwise distances...')
distances, rev_comp_errors = seqsum.summarize_distances(
seqs,
sample_size=args.num_samples,
per_site=True,
aligned=args.aligned,
ignore_gaps=(not args.count_gaps),
alphabet=alphabet,
do_full_alignment=args.msa,
full_alignment_out_path=full_alignment_out_path,
aligner_tools=aligner_tools,
full_aligner_tools=full_aligner_tools,
log_frequency=args.log_frequency)
log.info('Done!')
log.info('Writing mean distances to file...')
distances = sorted([(k, v) for k, v in iteritems(distances)],
key=lambda x: x[1].mean,
reverse=True)
mean_path = functions.get_new_path(
os.path.join(args.output_dir, 'seqvet-mean-distances.txt'))
with OpenFile(mean_path, 'w') as out:
out.write('seq_id\tmean_distance\n')
for (seq_id, dist) in distances:
out.write('{0}\t{1}\n'.format(seq_id, dist.mean))
log.info('Writing max distances to file...')
distances = sorted(distances, key=lambda x: x[1].maximum, reverse=True)
max_path = functions.get_new_path(
os.path.join(args.output_dir, 'seqvet-max-distances.txt'))
with OpenFile(max_path, 'w') as out:
out.write('seq_id\tmax_distance\n')
for (seq_id, dist) in distances:
out.write('{0}\t{1}\n'.format(seq_id, dist.maximum))
if rev_comp_errors:
rev_comp_errors = sorted(rev_comp_errors)
rce_set = set()
rce = []
for (s1, s2, d, drc) in rev_comp_errors:
pair = tuple(sorted([s1, s2]))
if pair in rce_set:
continue
rce_set.add(pair)
rce.append((pair[0], pair[1], d, drc))
log.info('Writing potential reverse-complement errors to file...')
path = functions.get_new_path(
os.path.join(args.output_dir,
'seqvet-reverse-complement-warnings.txt'))
with OpenFile(path, 'w') as out:
out.write('seq1\tseq2\tdistance\trev_comp_distance\n')
for (seq1, seq2, d, drc) in rce:
out.write('{0}\t{1}\t{2}\t{3}\n'.format(seq1, seq2, d, drc))