def main():
parser = argparse.ArgumentParser(
description="Build scaffolds from an 'orderings.bed' file")
parser.add_argument("agp",
metavar="<ragtag.correction.agp>",
type=str,
help="AGP v2.1 file produced by 'ragtag.py correct'")
parser.add_argument(
"query",
metavar="<query.fasta>",
type=str,
help="query fasta file to be scaffolded. must not be gzipped")
args = parser.parse_args()
agp_file = args.agp
query_file = args.query
x = pysam.FastaFile(query_file)
agp = AGPFile(agp_file)
# Iterate through the agp file
for line in agp.iterate_lines():
if line.is_gap:
raise ValueError("The AGP file should have no gaps.")
if line.orientation == "-":
raise ValueError("No sequences should have a '-' orientation.")
start, end = int(line.obj_beg) - 1, int(line.obj_end)
print(">" + line.comp)
print(x.fetch(line.obj, start, end))
def main():
parser = argparse.ArgumentParser(description="Rename FASTA records.", usage="ragtag_rename.py <seqs.fa> [-p PREFIX]")
parser.add_argument("fasta_fn", metavar="<seqs.fa>", default="", type=str, help="FASTA file (uncompressed or bgzipped)")
parser.add_argument("-p", metavar="STR", type=str, default="", help="prefix")
parser.add_argument("-o", metavar="PATH", type=str, default="ragtag.rename.agp", help="output AGP file path [./ragtag.rename.agp]")
args = parser.parse_args()
fasta_fn = args.fasta_fn
prefix = args.p
agp_fn = args.o
agp = AGPFile(agp_fn, "w")
record_idx = 0
fai = pysam.FastaFile(fasta_fn)
for reference in fai.references:
agp.add_seq_line(
prefix + "{0:08}".format(record_idx),
1,
fai.get_reference_length(reference),
"1",
"W",
reference,
1,
fai.get_reference_length(reference),
"+"
)
print(">" + prefix + "{0:08}".format(record_idx))
print(fai.fetch(reference))
record_idx += 1
agp.write()
fai.close()
def main():
parser = argparse.ArgumentParser(
description="Break corrected query sequences (objects) into components."
)
parser.add_argument("agp",
metavar="<ragtag.correction.agp>",
type=str,
help="AGP v2.1 file produced by 'ragtag.py correct'")
parser.add_argument(
"query",
metavar="<query.fasta>",
type=str,
help=
"query fasta file corresponding to objects in <ragtag.correction.agp> (can be uncompressed or bgzipped"
)
args = parser.parse_args()
agp_file = args.agp
query_file = args.query
fai = pysam.FastaFile(query_file)
agp = AGPFile(agp_file, mode="r")
# Iterate through the agp file
for line in agp.iterate_lines():
if line.is_gap:
raise ValueError("The AGP file should have no gaps.")
if line.orientation == "-":
raise ValueError("No sequences should have a '-' orientation.")
start, end = int(line.obj_beg) - 1, int(line.obj_end)
print(">" + line.comp)
print(fai.fetch(line.obj, start, end))
fai.close()
def main():
parser = argparse.ArgumentParser(
description="Build sequences in FASTA format from an AGP v2.1 file.",
usage="ragtag.py agp2fa <scaffolds.agp> <components.fasta>")
parser.add_argument("agp",
metavar="<scaffolds.agp>",
nargs='?',
default="",
type=str,
help="AGP v2.1 file")
parser.add_argument(
"components",
metavar="<components.fasta>",
nargs='?',
default="",
type=str,
help="component FASTA file (can be uncompressed or bgzipped)")
args = parser.parse_args()
if not args.agp or not args.components:
parser.print_help()
sys.exit()
agp_file = args.agp
components_file = args.components
fai = pysam.FastaFile(components_file)
agp = AGPFile(agp_file, mode="r")
# Iterate over the lines of the AGP file
prev_obj = None
is_first = True
for agp_line in agp.iterate_lines():
if agp_line.obj != prev_obj:
if is_first:
print(">" + agp_line.obj)
is_first = False
else:
print("\n>" + agp_line.obj)
prev_obj = agp_line.obj
if agp_line.is_gap:
sys.stdout.write("N" * agp_line.gap_len)
else:
if agp_line.orientation == "-":
sys.stdout.write(
reverse_complement(
fai.fetch(agp_line.comp, agp_line.comp_beg - 1,
agp_line.comp_end)))
else:
sys.stdout.write(
fai.fetch(agp_line.comp, agp_line.comp_beg - 1,
agp_line.comp_end))
# End the FASTA file with a newline
sys.stdout.write("\n")
fai.close()
def main():
parser = argparse.ArgumentParser(description="Calculate scaffolding statistics")
parser.add_argument("agp", nargs='?', default="", metavar="<ragtag.scaffolds.agp>", type=str, help="RagTag scaffolding AGP file")
parser.add_argument("confidence", nargs='?', default="", metavar="<ragtag.confidence.txt>", type=str, help="RagTag scaffolding confidence scores file")
args = parser.parse_args()
if not args.agp or not args.confidence:
parser.print_help()
sys.exit()
agp_file = args.agp
confidence_file = args.confidence
placed_bp = 0
placed_seq = 0
unplaced_bp = 0
unplaced_seq = 0
gap_bp = 0
gap_seq = 0
allowed_seq_types = {"A", "D", "F", "G", "O", "P", "W"}
allowed_gap_types = {"N", "U"}
# Get the set of placed sequences from the confidence scores file
placed_seqs = set()
with open(confidence_file, "r") as f:
f.readline() # discard header
for line in f:
header, g_score, l_score, o_score = line.rstrip().split("\t")
placed_seqs.add(header)
# Iterate through the AGP file
agp = AGPFile(agp_file, mode="r")
for line in agp.iterate_lines():
if line.is_gap:
gap_bp += line.gap_len
gap_seq += 1
else:
seq_len = line.comp_end - (line.comp_beg - 1)
if line.comp in placed_seqs:
placed_bp += seq_len
placed_seq += 1
else:
unplaced_bp += seq_len
unplaced_seq += 1
print("placed_sequences\tplaced_bp\tunplaced_sequences\tunplaced_bp\tgap_bp\tgap_sequences")
print("\t".join([
str(placed_seq),
str(placed_bp),
str(unplaced_seq),
str(unplaced_bp),
str(gap_bp),
str(gap_seq)
]))
def sub_update(gff_file, agp_file):
# Make a dictionary associating each original sequence with an interval tree of component sequences
trans = defaultdict(IntervalTree)
agp = AGPFile(agp_file, mode="r")
for agp_line in agp.iterate_lines():
# Check that the agp file looks correct for this task
if agp_line.orientation == "-":
raise ValueError(
"The placement BED file is not formatted correctly. No sequences should be reverse complemented for misassembly correction."
)
if not agp_line.comp_type == "W":
raise ValueError(
"The placement BED file is not formatted correctly. All lines should be WGS contig (W)."
)
if agp_line.is_gap:
raise ValueError(
"There should be no gaps in the correction AGP file.")
start, end = agp_line.obj_beg - 1, agp_line.obj_end
trans[agp_line.obj][start:end] = agp_line.comp
# Iterate through the gff intervals and update them according to trans
with open(gff_file, "r") as f:
for line in f:
line = line.rstrip()
if line.startswith("#"):
print(line) # Print this comment line
else:
fields = line.split("\t")
h, s, e = fields[0], int(fields[3]), int(fields[4])
s -= 1 # Keep everything zero-indexed
if h not in trans:
raise ValueError("Inconsistent input files.")
ovlps = trans[h][s:e]
if len(ovlps) > 1:
raise ValueError(
"%s:%d-%d in the gff file overlaps two sub sequences in the placement file. Make sure to run 'ragtag.py correct' with '--gff'"
% (h, s, e))
if len(ovlps) < 1:
raise ValueError(
"The placement BED file is not formatted correctly.")
# Get the data from the overlapping interval and print the new line
o = list(ovlps)[0]
new_s = s - o.begin
new_e = e - o.begin
fields[0] = o.data
fields[3] = str(new_s +
1) # back to one-based indexing for gff format
fields[4] = str(new_e)
print("\t".join(fields))
def write_breaks(out_file, query_file, ctg_breaks, overwrite, remove_suffix):
""" Write the intermediate file for contig breaks in AGP v2.1 format."""
# Check if the output file already exists
if os.path.isfile(out_file):
if not overwrite:
log("Retaining pre-existing file: " + out_file)
return
else:
log("Overwriting pre-existing file: " + out_file)
fai = pysam.FastaFile(query_file)
all_q_seqs = sorted(fai.references)
agp = AGPFile(out_file, mode="w")
agp.add_pragma()
agp.add_comment("# AGP created by RagTag {}".format(get_ragtag_version()))
for q in all_q_seqs:
# Check if this sequence was broken during misassembly correction
if q not in ctg_breaks:
# Add suffix to query header, unless otherwise requested
unchanged_comp_header = q
if not remove_suffix:
unchanged_comp_header = q + ":0" + "-" + str(
fai.get_reference_length(q)) + "(+)"
agp.add_seq_line(q, "1", str(fai.get_reference_length(q)), "1",
"W", unchanged_comp_header, "1",
str(fai.get_reference_length(q)), "+")
else: # This query sequence was broken
pid = 1
sorted_breaks = sorted(ctg_breaks[q])
start = 0
for i in sorted_breaks:
agp.add_seq_line(q, str(start + 1), str(i), str(pid), "W",
q + ":" + str(start) + "-" + str(i) + "(+)",
"1", str(i - start), "+")
start = i
pid += 1
# Add one line for the last interval
agp.add_seq_line(
q, str(start + 1), str(fai.get_reference_length(q)), str(pid),
"W", q + ":" + str(start) + "-" +
str(fai.get_reference_length(q)) + "(+)", "1",
str(fai.get_reference_length(q) - start), "+")
log("Writing: " + out_file)
agp.write()
fai.close()
def sup_update(gff_file, agp_file):
# Make a dictionary associating each original sequence with the destination sequence
trans = {}
strands = {}
seq_lens = {}
agp = AGPFile(agp_file, mode="r")
for agp_line in agp.iterate_lines():
if not agp_line.is_gap:
start, end = agp_line.obj_beg - 1, agp_line.obj_end
trans[agp_line.comp] = (start, end, agp_line.obj)
strands[agp_line.comp] = agp_line.orientation
seq_lens[agp_line.comp] = end - start
# Iterate through the gff intervals and update them according to trans
with open(gff_file, "r") as f:
for line in f:
line = line.rstrip()
if line.startswith("#"):
print(line) # Print this comment line
else:
fields = line.split("\t")
h, s, e, st = fields[0], int(fields[3]), int(
fields[4]), fields[6]
s -= 1 # Keep everything zero-indexed
if h not in trans:
print()
print(line)
raise ValueError("Inconsistent input files.")
# Check if the original sequence has been reverse complemented
if strands[h] == "-":
l = seq_lens[h]
s, e = l - e, l - s
if st == "+":
st = "-"
else:
st = "+"
new_s = trans[h][0] + s
new_e = trans[h][0] + e
fields[0] = trans[h][2]
fields[3] = str(new_s +
1) # back to one-based indexing for gff format
fields[4] = str(new_e)
fields[6] = st
print("\t".join(fields))
def main():
parser = argparse.ArgumentParser(
description="Build sequences in FASTA format from an AGP v2.1 file. ")
parser.add_argument("agp",
metavar="<scaffolds.agp>",
type=str,
help="AGP v2.1 file")
parser.add_argument(
"components",
metavar="<components.fasta>",
type=str,
help=
"FASTA file with component sequences to be scaffolded. must not be gzipped"
)
args = parser.parse_args()
agp_file = args.agp
components_file = args.components
fai = pysam.FastaFile(components_file)
agp = AGPFile(agp_file)
# Iterate over the lines of the AGP file
prev_obj = None
is_first = True
for agp_line in agp.iterate_lines():
if agp_line.obj != prev_obj:
if is_first:
print(">" + agp_line.obj)
is_first = False
else:
print("\n>" + agp_line.obj)
prev_obj = agp_line.obj
if agp_line.is_gap:
sys.stdout.write("N" * agp_line.gap_len)
else:
if agp_line.orientation == "-":
sys.stdout.write(reverse_complement(fai.fetch(agp_line.comp)))
else:
sys.stdout.write(fai.fetch(agp_line.comp))
# End the FASTA file with a newline
sys.stdout.write("\n")
def main():
parser = argparse.ArgumentParser(description="Check AGP v2.1 files for validity.", usage="ragtag.py agpcheck <asm1.agp> [<asm2.agp> ... <asmN.agp>]")
parser.add_argument("agp", metavar="<asm1.agp> [<asm2.agp> ... <asmN.agp>]", nargs='+', default=[], type=str, help="AGP v2.1 files")
DISCLAIMER = """
DISCLAIMER:
This utility performs most (but not all) checks necessary to validate an
AGP v2.1 file: https://www.ncbi.nlm.nih.gov/assembly/agp/AGP_Specification/
Please additionally use the NCBI AGP validator for robust
validation: https://www.ncbi.nlm.nih.gov/assembly/agp/AGP_Validation/
"""
args = parser.parse_args()
print(DISCLAIMER)
agp_file_list = [os.path.abspath(i) for i in args.agp]
for agp_file in agp_file_list:
print()
log("INFO", "Checking {} ...".format(agp_file))
agp = AGPFile(agp_file, mode="r")
for _ in agp.iterate_lines():
pass
log("INFO", "Check for {} is complete with no errors.".format(agp_file))
def write_agp(self, agp_fn, ref_fn, add_suffix_to_unplaced=False):
"""
Write the AGP file implied by the scaffold graph
:param agp_fn: AGP file name
:param ref_fn: reference FASTA file name
:param add_suffix_to_unplaced: add "_RagTag" to unscaffolded sequences
"""
used_components = set()
used_edges = set()
obj_header_idx = -1
agp = AGPFile(agp_fn, "w")
agp.add_pragma()
agp.add_comment("# AGP created by RagTag {}".format(get_ragtag_version()))
while True:
# Find a starting node
from_node = None
to_node = None
cur_ref = None
for u, v in sorted(self.edges):
if (u, v) not in used_edges:
u_base = u[:-2]
u_degree = 0
if u_base + "_b" in self.nodes:
u_degree += self.graph.degree[u_base + "_b"]
if u_base + "_e" in self.nodes:
u_degree += self.graph.degree[u_base + "_e"]
assert u_degree in {2, 4}
# Check if we have found a starting target sequence
if u_degree == 2:
cur_ref = u_base
from_node = u
to_node = v
used_edges.add((u, v))
used_edges.add((v, u))
break
# If we haven't found a new starting target sequence, we are done
if from_node is None:
break
# Initialize this object
obj_header_idx += 1
obj_header = "scf" + "{0:08}".format(obj_header_idx)
obj_pos = 0
obj_pid = 1
# Process the first target sequence
cur_ref_len = self.component_lens[cur_ref]
cur_ref_strand = "+"
if from_node.endswith("_b"):
cur_ref_strand = "-"
agp.add_seq_line(obj_header, obj_pos+1, obj_pos+cur_ref_len, obj_pid, "W", cur_ref, 1, cur_ref_len, cur_ref_strand)
obj_pos += cur_ref_len
obj_pid += 1
used_components.add(cur_ref)
# Process the remaining sequences.
next_edge_exists = True
while next_edge_exists:
# Process the patch
patch_aln = self.graph[from_node][to_node]["alignment"]
patch_query = patch_aln.query
patch_strand = "+"
if patch_aln.strand:
patch_strand = "-"
patch_len = patch_aln.their_query_start - patch_aln.my_query_end
if patch_len > 0:
if patch_aln.is_gap:
agp.add_gap_line(obj_header, obj_pos+1, obj_pos+patch_len, obj_pid, "N", patch_len, "scaffold", "yes", "align_genus")
else:
agp.add_seq_line(obj_header, obj_pos+1, obj_pos+patch_len, obj_pid, "W", patch_query, patch_aln.my_query_end+1, patch_aln.their_query_start, patch_strand)
used_components.add(patch_query)
obj_pos += patch_len
obj_pid += 1
# Next, process the reference sequence
comp_start = min(0, patch_len)
cur_ref = to_node[:-2]
cur_ref_len = self.component_lens[cur_ref]
cur_ref_strand = "+"
if to_node.endswith("_e"):
cur_ref_strand = "-"
agp.add_seq_line(obj_header, obj_pos+1, obj_pos+(cur_ref_len + comp_start), obj_pid, "W", cur_ref, 1+(-1*comp_start), cur_ref_len, cur_ref_strand)
obj_pos += cur_ref_len + comp_start
obj_pid += 1
used_components.add(cur_ref)
# Look for the next edge
from_node = to_node[:-2] + "_b"
if to_node.endswith("_b"):
from_node = to_node[:-2] + "_e"
if from_node in self.graph.nodes:
next_nodes = set(self.graph[from_node])
assert len(next_nodes) == 1
to_node = next_nodes.pop()
used_edges.add((from_node, to_node))
used_edges.add((to_node, from_node))
else:
next_edge_exists = False
# Write unplaced reference sequences
fai = pysam.FastaFile(ref_fn)
all_ref_seqs = set(fai.references)
fai.close()
remaining_components = all_ref_seqs - used_components
for c in sorted(remaining_components):
agp.add_seq_line(
c + "_RagTag" * add_suffix_to_unplaced,
"1",
str(self.component_lens[c]),
"1",
"W",
c,
"1",
str(self.component_lens[c]),
"+"
)
agp.write()
def _get_assembly_points(self, agp, weight):
"""
Find all adjacencies defined in an AGP file
:param agp: An AGP file defining sequence adjacencies
:param weight: The weight to assign to each adjacency
"""
comps = set()
prev_obj = ""
seq1 = ""
strand1 = ""
# Gap info
gap_count = 0
prev_agp_known = None
prev_gap_size = 0
prev_gap_type = None
prev_linkage = ""
prev_evidence = ""
# Iterate over the AGP file and yield assembly points
agp_file = AGPFile(agp)
for agp_line in agp_file.iterate_lines():
if not agp_line.is_gap:
# Add this component to our master list
if agp_line.comp not in self.component_lens:
raise RuntimeError("{} is in {} but not {}.".format(agp_line.comp, agp, self.components_fasta_fname))
comps.add(agp_line.comp)
comp_len = agp_line.comp_end
if comp_len < self.get_component_len(agp_line.comp):
raise RuntimeError("only complete components can be added to the graph.")
if comp_len > self.get_component_len(agp_line.comp):
raise RuntimeError("inconsistent component lengths: {} bp in {} and {} bp in {}". format(comp_len, agp, self.get_component_len(agp_line.comp), self.components_fasta_fname))
if agp_line.obj == prev_obj:
# Check if these components are bookended (no gap in between)
if not gap_count:
prev_evidence = "bookend"
# Check if two consecutive gaps preceded this component
if gap_count > 1:
raise ValueError("Consecutive gaps in the AGP file are not currently supported.")
yield AssemblyPoint(
seq1,
strand1,
agp_line.comp,
agp_line.orientation,
weight,
agp,
prev_agp_known,
prev_gap_size,
prev_gap_type,
prev_linkage,
prev_evidence
)
# Set this component as the previous component
seq1 = agp_line.comp
strand1 = agp_line.orientation
gap_count = 0
prev_agp_known = None
prev_gap_size = 0
prev_gap_type = None
prev_linkage = ""
prev_evidence = ""
else:
seq1 = agp_line.comp
strand1 = agp_line.orientation
prev_obj = agp_line.obj
gap_count = 0
prev_agp_known = None
prev_gap_size = 0
prev_gap_type = None
prev_linkage = ""
prev_evidence = ""
else:
if agp_line.obj == prev_obj:
gap_count += 1
prev_agp_known = True if agp_line.comp_type == "N" else False
prev_gap_size = agp_line.gap_len
prev_gap_type = agp_line.gap_type
prev_linkage = True if agp_line.linkage == "yes" else False
prev_evidence = agp_line.linkage_evidence
if comps != self.components:
raise ValueError("Input AGPs do not have the same set of components.")
def write_orderings(out_agp_file, out_confidence_file, query_file,
ordering_dict, ctg_dict, gap_dict, gap_type_dict,
make_chr0, overwrite, add_suffix):
# Check if the output file already exists
if os.path.isfile(out_agp_file):
if not overwrite:
log("Retaining pre-existing file: " + out_agp_file)
return
else:
log("Overwriting pre-existing file: " + out_agp_file)
# Proceed with writing the intermediate output
placed_seqs = set()
all_out_cs_lines = [] # For confidence scores
agp = AGPFile(out_agp_file, mode="w")
agp.add_pragma()
agp.add_comment("# AGP created by RagTag {}".format(get_ragtag_version()))
# Go through the reference sequences in sorted order
sorted_ref_headers = sorted(list(ordering_dict.keys()))
for ref_header in sorted_ref_headers:
pid = 1
pos = 0
new_ref_header = ref_header + "_RagTag"
q_seqs = ordering_dict[ref_header]
gap_seqs = gap_dict[ref_header]
gap_types = gap_type_dict[ref_header]
# Iterate through the query sequences for this reference header
for i in range(len(q_seqs)):
out_agp_line = []
out_cs_line = []
q = q_seqs[i][2]
placed_seqs.add(q)
qlen = ctg_dict[q].query_len
strand = ctg_dict[q].orientation
gc, lc, oc = ctg_dict[q].grouping_confidence, ctg_dict[
q].location_confidence, ctg_dict[q].orientation_confidence
out_agp_line.append(new_ref_header)
out_agp_line.append(str(pos + 1))
pos += qlen
out_agp_line.append(str(pos))
out_agp_line.append(str(pid))
out_agp_line.append("W")
out_agp_line.append(q)
out_agp_line.append("1")
out_agp_line.append(str(ctg_dict[q].query_len))
out_agp_line.append(strand)
# Save the confidence score info
out_cs_line.append(q)
out_cs_line.append(str(gc))
out_cs_line.append(str(lc))
out_cs_line.append(str(oc))
agp.add_seq_line(*out_agp_line)
all_out_cs_lines.append("\t".join(out_cs_line))
pid += 1
if i < len(gap_seqs):
# Print the gap line
out_agp_line = []
out_agp_line.append(new_ref_header)
out_agp_line.append(str(pos + 1))
pos += gap_seqs[i]
out_agp_line.append(str(pos))
out_agp_line.append(str(pid))
gap_type = gap_types[i]
out_agp_line.append(gap_type)
out_agp_line.append(str(gap_seqs[i]))
out_agp_line.append("scaffold")
out_agp_line.append("yes")
out_agp_line.append("align_genus")
pid += 1
agp.add_gap_line(*out_agp_line)
# Write unplaced sequences
fai = pysam.FastaFile(query_file)
all_seqs = set(fai.references)
unplaced_seqs = sorted(list(all_seqs - placed_seqs))
if unplaced_seqs:
if make_chr0:
pos = 0
pid = 1
new_ref_header = "Chr0_RagTag"
for q in unplaced_seqs:
out_agp_line = []
qlen = fai.get_reference_length(q)
out_agp_line.append(new_ref_header)
out_agp_line.append(str(pos + 1))
pos += qlen
out_agp_line.append(str(pos))
out_agp_line.append(str(pid))
out_agp_line.append("W")
out_agp_line.append(q)
out_agp_line.append("1")
out_agp_line.append(str(qlen))
out_agp_line.append("+")
agp.add_seq_line(*out_agp_line)
pid += 1
# Now for the gap, since we are making a chr0
out_agp_line = []
out_agp_line.append(new_ref_header)
out_agp_line.append(str(pos + 1))
pos += 100
out_agp_line.append(str(pos))
out_agp_line.append(str(pid))
out_agp_line.append("U")
out_agp_line.append("100")
out_agp_line.append("contig")
out_agp_line.append("no")
out_agp_line.append("na")
agp.add_gap_line(*out_agp_line)
pid += 1
# Remove the final unecessary gap
agp.pop_agp_line()
else:
# List the unplaced contigs individually
for q in unplaced_seqs:
out_agp_line = []
qlen = fai.get_reference_length(q)
if add_suffix:
out_agp_line.append(q + "_RagTag")
else:
out_agp_line.append(q)
out_agp_line.append("1")
out_agp_line.append(str(qlen))
out_agp_line.append("1")
out_agp_line.append("W")
out_agp_line.append(q)
out_agp_line.append("1")
out_agp_line.append(str(qlen))
out_agp_line.append("+")
agp.add_seq_line(*out_agp_line)
agp.write()
fai.close()
# Write the confidence scores
with open(out_confidence_file, "w") as f:
f.write(
"query\tgrouping_confidence\tlocation_confidence\torientation_confidence\n"
)
f.write("\n".join(all_out_cs_lines) + "\n")
def write_agp_solution(cover_graph, scaffold_graph, agp_fname, gap_func="MIN", add_suffix_to_unplaced=False):
"""
Here, we work with two graphs: A cover_graph and a scaffold_graph. A covergrpah defines a solution to the scaffold
graph, and nodes from the same component are connected for convenience.
We use the scaffold_graph for any original scaffold_graph info/functionality
"""
if not isinstance(scaffold_graph, ScaffoldGraphBase):
raise TypeError("scaffold_graph must be an instance of ScaffoldGraph")
placed_components = set()
# Iterate over each connected component
agp = AGPFile(agp_fname, mode="w")
agp.add_pragma()
agp.add_comment("# AGP created by RagTag {}".format(get_ragtag_version()))
# Iterate through the connected components
for i, cc in enumerate(nx.connected_components(G=cover_graph)):
# Sort the list of nodes for deterministic output
cc = sorted(list(cc))
obj_header = "scf" + "{0:08}".format(i+1) + "_RagTag"
current_node = None
# Iterate over each node in the connected component until we find a node with degree=1
for node in cc:
if cover_graph.degree[node] == 1:
current_node = node
break
assert current_node is not None
# Starting with the degree=1 node, build the AGP object from nodes in the path.
visited_nodes = {current_node}
degree = 0
obj_id = 1
obj_pos = 0
# Traverse the component until we find the other end node
while degree != 1:
conn_nodes = set(cover_graph.neighbors(current_node))
next_node = (conn_nodes - visited_nodes).pop()
degree = cover_graph.degree[next_node]
comp_len = scaffold_graph.get_component_len(next_node[:-2])
# Check if this is an intra or inter sequence edge
orientation = "+"
if next_node[:-2] == current_node[:-2]:
if next_node.endswith("_b"):
orientation = "-"
assert current_node.endswith("_e")
agp.add_seq_line(
obj_header,
str(obj_pos + 1),
str(obj_pos + comp_len),
str(obj_id),
"W",
next_node[:-2],
"1",
str(comp_len),
orientation
)
obj_pos += comp_len
placed_components.add(next_node[:-2])
else:
# Organize the gap info
adjacency_data = scaffold_graph[current_node][next_node]
# AGP Column 5
all_is_known_gap_size = adjacency_data["is_known_gap_size"]
comp_type = "N" if any(all_is_known_gap_size) else "U"
# AGP column 6b
gap_size = 100
all_gap_sizes = adjacency_data["gap_size"]
fltrd_gap_sizes = [all_gap_sizes[i] for i in range(len(all_gap_sizes)) if all_is_known_gap_size[i]]
if fltrd_gap_sizes:
if len(fltrd_gap_sizes) == 1:
gap_size = fltrd_gap_sizes[0]
else:
gap_size = get_gap_size(fltrd_gap_sizes, gap_func)
# AGP column 7b
all_gap_types = set(adjacency_data["gap_type"])
gap_type = "scaffold"
if len(all_gap_types) == 1:
gap_type = all_gap_types.pop()
# AGP column 8b
has_linkage = "yes" if any(adjacency_data["linkage"]) else "no"
# AGP column 9b
all_evidences = set(adjacency_data["linkage_evidence"])
linkage_evidence = "na"
if has_linkage == "yes":
if "na" in all_evidences:
all_evidences.remove("na")
linkage_evidence = ";".join([str(i) for i in all_evidences])
agp.add_gap_line(
obj_header,
str(obj_pos + 1),
str(obj_pos + gap_size),
str(obj_id),
comp_type,
str(gap_size),
gap_type,
has_linkage,
linkage_evidence
)
obj_pos += gap_size
obj_id += 1
visited_nodes.add(next_node)
current_node = next_node
# Write all unplaced contigs
remaining_components = scaffold_graph.components - placed_components
for c in remaining_components:
agp.add_seq_line(
c + "_RagTag" * add_suffix_to_unplaced,
"1",
str(scaffold_graph.get_component_len(c)),
"1",
"W",
c,
"1",
str(scaffold_graph.get_component_len(c)),
"+"
)
agp.write()
def lens_from_agp(fname):
agp_file = AGPFile(fname, mode="r")
return [obj.obj_len for obj in agp_file.iterate_objs()]
def main():
parser = argparse.ArgumentParser(description='Split sequencs at gaps',
usage="ragtag.py splitasm <asm.fa>")
parser.add_argument("asm",
metavar="<asm.fa>",
default="",
type=str,
help="assembly fasta file (uncompressed or bgzipped)")
parser.add_argument("-n",
metavar="INT",
type=int,
default=0,
help="minimum gap size [0]")
parser.add_argument("-o",
metavar="PATH",
type=str,
default="ragtag.splitasm.agp",
help="output AGP file path [./ragtag.splitasm.agp]")
# Parse the command line arguments
args = parser.parse_args()
if not args.asm:
parser.print_help()
print("\n** The assembly FASTA file is required **")
sys.exit()
asm_fn = args.asm
min_gap_size = args.n
agp_fn = args.o
# Initialize the AGP file
agp = AGPFile(agp_fn, mode="w")
agp.add_pragma()
agp.add_comment("# AGP created by RagTag {}".format(get_ragtag_version()))
# Process the FASTA file
new_header_idx = 0
fai = pysam.FastaFile(asm_fn)
for header in sorted(fai.references):
seq = fai.fetch(header).upper()
seq_len = fai.get_reference_length(header)
gap_coords = [(i.start(), i.end()) for i in re.finditer(r'N+', seq)
if i.end() - i.start() > min_gap_size]
if not gap_coords:
new_header = "seq{0:08}".format(new_header_idx)
new_header_idx += 1
agp.add_seq_line(header, "1", seq_len, "1", "W", new_header, "1",
seq_len, "+")
else:
gap_coords.append((seq_len, seq_len + 1))
pid = 1
if gap_coords[0][0]:
# The sequence doesn't start with a gap
new_header = "seq{0:08}".format(new_header_idx)
agp.add_seq_line(header, "1", str(gap_coords[0][0]),
str(pid), "W", new_header, "1",
str(gap_coords[0][0]), "+")
new_header_idx += 1
pid += 1
for i in range(1, len(gap_coords)):
# Add the gap line
gap_start, gap_end = gap_coords[i - 1][0], gap_coords[i - 1][1]
gap_len = gap_end - gap_start
agp.add_gap_line(header, str(gap_start + 1), str(gap_end),
str(pid), "N", str(gap_len), "scaffold",
"yes", "align_genus")
pid += 1
# Add the sequence line
obj_start, obj_end = gap_coords[i - 1][1], gap_coords[i][0]
comp_len = obj_end - obj_start
new_header = "seq{0:08}".format(new_header_idx)
if gap_coords[i - 1][1] != seq_len:
agp.add_seq_line(header, str(obj_start + 1), obj_end, pid,
"W", new_header, "1", str(comp_len), "+")
new_header_idx += 1
pid += 1
agp.write()
# Iterate over the AGP file and print the sequences
agp = AGPFile(agp_fn, mode="r")
for line in agp.iterate_lines():
if not line.is_gap:
obj, comp, obj_beg, obj_end = line.obj, line.comp, line.obj_beg, line.obj_end
print(">" + comp)
print(fai.fetch(obj, obj_beg - 1, obj_end))
fai.close()