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#!/usr/bin/env python2.7
#(c) 2013-2014 by Authors
#This file is a part of Ragout program.
#Released under the BSD license (see LICENSE file)
"""
This script checks correctness of Ragout results
if a 'true' reference is available
"""
from __future__ import print_function
from __future__ import absolute_import
import sys
import os
from collections import namedtuple
from itertools import product
import subprocess
from utils.common import AlignmentRow, AlignmentColumn
from utils.common import (filter_by_coverage, join_collinear,
group_by_chr, get_order)
from six.moves import map
from six.moves import range
Scaffold = namedtuple("Scaffold", ["name", "contigs"])
Contig = namedtuple("Contig", ["name", "sign"])
MINIMAP_BIN = "minimap2"
def read_fasta_dict(filename):
"""
Reads fasta file into dictionary. Also preforms some validation
"""
#logger.debug("Reading contigs file")
header = None
seq = []
fasta_dict = {}
try:
with open(filename, "r") as f:
for lineno, line in enumerate(f):
line = line.strip()
if line.startswith(">"):
if header:
fasta_dict[header] = "".join(seq)
seq = []
header = line[1:].split(" ")[0]
else:
seq.append(line)
if header and len(seq):
fasta_dict[header] = "".join(seq)
except IOError as e:
raise Exception(e)
return fasta_dict
def write_fasta_dict(fasta_dict, filename):
"""
Writes dictionary with fasta to file
"""
with open(filename, "w") as f:
for header in sorted(fasta_dict):
f.write(">{0}\n".format(header))
for i in range(0, len(fasta_dict[header]), 60):
f.write(fasta_dict[header][i:i + 60] + "\n")
def parse_links_file(filename):
scaffolds = []
def add_contig(string):
name = string.replace("=", "_") #fix for nucmer
without_sign = name[1:].strip()
sign = 1 if name[0] == "+" else -1
scaffolds[-1].contigs.append(Contig(without_sign, sign))
with open(filename, "r") as f:
for line in f:
line = line.strip()
if not line or line.startswith("--") or line.startswith("sequence"):
continue
if line[0] not in ["+", "-"]:
scaffolds.append(Scaffold(line, []))
else:
left_cont = line.split()[0]
add_contig(left_cont)
#add_contig(right_cont)
return scaffolds
def read_paf(filename):
alignment = []
for line in open(filename, "r"):
line = line.strip()
if not len(line):
continue
vals = line.split("\t")
s_ref, e_ref = int(vals[7]), int(vals[8])
s_qry, e_qry = int(vals[2]), int(vals[3])
len_ref, len_qry = int(vals[6]), int(vals[1])
ref_id, qry_id = vals[5], vals[0]
ref_strand = 1
qry_strand = 1
if vals[4] == "-":
qry_strand = -1
ref_row = AlignmentRow(s_ref, e_ref, ref_strand, len_ref, ref_id)
qry_row = AlignmentRow(s_qry, e_qry, qry_strand, len_qry, qry_id)
alignment.append(AlignmentColumn(ref_row, qry_row))
return alignment
def get_alignment(scaffolds, contigs_file, reference_file):
query_file = "_minimap-query.fasta"
query_fasta = {}
contigs_fasta = read_fasta_dict(contigs_file)
for scf in scaffolds:
for ctg in scf.contigs:
if "[" not in ctg.name:
query_fasta[ctg.name] = contigs_fasta[ctg.name]
else:
bracket = ctg.name.index("[")
start, end = list(map(int, ctg.name[bracket + 1: -1].split(":")))
seq_name = ctg.name[:bracket]
query_fasta[ctg.name] = contigs_fasta[seq_name][start:end]
write_fasta_dict(query_fasta, query_file)
#run minimap here
minimap_file = "_minimap.paf"
subprocess.check_call([MINIMAP_BIN, reference_file, query_file, "-x", "asm5",
"--secondary=no", "-t", "8"], stdout=open(minimap_file, "w"))
aln = read_paf(minimap_file)
os.remove(query_file)
os.remove(minimap_file)
return aln
def gap_count(lst_1, lst_2):
if not lst_1 or not lst_2:
return 0
gaps = sys.maxsize
for i, j in product(lst_1, lst_2):
gaps = min(gaps, abs(i.index - j.index) - 1)
return gaps
def agreement_ord(increasing, lst_1, lst_2, chr_len_dict):
if not lst_1 or not lst_2:
return True
for i, j in product(lst_1, lst_2):
same_chr = i.chr == j.chr
if not same_chr:
continue
chr_len = chr_len_dict[i.chr]
lower = chr_len * 0.2
higher = chr_len * 0.8
over_zero = ((increasing and i.coord > higher and j.coord < lower) or
(not increasing and i.coord < lower and j.coord > higher))
if ((j.index > i.index) == increasing and
abs(i.coord - j.coord) < chr_len / 1) or over_zero:
return True
return False
def agreement_strands(lst_1, lst_2, increasing):
incr_sign = 1 if increasing else -1
for i, j in product(lst_1, lst_2):
if i == j and i == incr_sign:
return True
return False
def do_job(links_file, contigs_file, reference_file):
scaffolds = parse_links_file(links_file)
alignment = get_alignment(scaffolds, contigs_file, reference_file)
alignment = filter_by_coverage(alignment, 0.45)
alignment = join_collinear(alignment)
entry_ord, chr_len, contig_len = get_order(alignment)
total_breaks = 0
total_gaps = 0
total_contigs = 0
for s in scaffolds:
print("\n>" + s.name)
prev_aln = []
prev_strand = None
increasing = None
breaks = []
for contig in s.contigs:
miss_ord = False
miss_strand = False
#flipping alignments
for hit in entry_ord[contig.name]:
if contig.sign < 0:
hit.sign = -hit.sign
#checking order
if prev_aln:
if increasing is not None:
if not agreement_ord(increasing, prev_aln,
entry_ord[contig.name], chr_len):
increasing = None
breaks.append(contig.name)
total_breaks += 1
miss_ord = True
elif len(entry_ord[contig.name]) == 1 and len(prev_aln) == 1:
increasing = (entry_ord[contig.name][0].index >
prev_aln[0].index)
#checking strand
cur_strand = [h.sign for h in entry_ord[contig.name]]
if not miss_ord and prev_strand and cur_strand:
if not agreement_strands(prev_strand, cur_strand, increasing):
breaks.append(contig.name)
total_breaks += 1
miss_strand = True
increasing = None
if gap_count(prev_aln, entry_ord[contig.name]) > 0:
total_gaps += 1
#only if this contig has alignments
if entry_ord[contig.name]:
prev_aln = entry_ord[contig.name]
prev_strand = cur_strand
#output
sign = "+" if contig.sign > 0 else "-"
pos_list = list(map(str, entry_ord[contig.name]))
pos_list_str = (str(pos_list) if len(pos_list) < 5 else
str(pos_list[:5]) + "...")
print("{0}{1}\t\t{2}\t{3}".format(sign, contig.name,
contig_len[contig.name], pos_list_str),
end="")
print("\t<<<order" if miss_ord else "", end="")
print("\t<<<strand" if miss_strand else "", end="")
print("")
total_contigs += 1
###
print("\tmiss-ordered: ", len(breaks))
print("\nTotal miss-ordered:", total_breaks)
print("Total gaps:", total_gaps)
print("Total contigs:", total_contigs)
print("Total scaffolds:", len(scaffolds))
def main():
if len(sys.argv) != 4:
print("Usage: verify-minimap links contigs reference")
return 1
do_job(sys.argv[1], sys.argv[2], sys.argv[3])
if __name__ == "__main__":
main()
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