def coding_regions(self):
coding_regions = {}
annotations = list(
annotation.read_annotations(join(self.dirname, 'reference.gff')))
annotation.link_up_annotations(annotations)
for item in annotations:
if item.type == "CDS":
[mrna] = item.parents
[gene] = mrna.parents
name = gene.get_id()
if name not in coding_regions:
coding_regions[name] = []
coding_regions[name].append(item)
for gene in annotations:
if gene.type != "gene": continue
name = gene.get_id()
if name not in coding_regions:
coding_region = gene.three_prime()
coding_region.attr = {"ID": name}
else:
items = coding_regions[name]
coding_region = annotation.Annotation(
type="coding_region",
seqid=items[0].seqid,
strand=items[0].strand,
start=min(item2.start for item2 in items),
end=max(item2.end for item2 in items),
attr={"ID": name},
)
coding_regions[name] = coding_region
return coding_regions
def coding_regions(self):
coding_regions = { }
annotations = list(annotation.read_annotations(join(self.dirname,'reference.gff')))
annotation.link_up_annotations(annotations)
for item in annotations:
if item.type == "CDS":
[ mrna ] = item.parents
[ gene ] = mrna.parents
name = gene.get_id()
if name not in coding_regions:
coding_regions[name] = [ ]
coding_regions[name].append(item)
for gene in annotations:
if gene.type != "gene": continue
name = gene.get_id()
if name not in coding_regions:
coding_region = gene.three_prime()
coding_region.attr = { "ID" : name }
else:
items = coding_regions[name]
coding_region = annotation.Annotation(
type = "coding_region",
seqid = items[0].seqid,
strand= items[0].strand,
start = min(item2.start for item2 in items),
end = max(item2.end for item2 in items),
attr = { "ID" : name },
)
coding_regions[name] = coding_region
return coding_regions
def get_genes(items, extractions, log):
annotation.link_up_annotations(items)
log.log("Content of downloaded GFF file (type:biotype ... paths):\n")
show_tree(items, log)
log.log("\n")
genes = list()
for gene_type,gene_biotype,transcript_type,transcript_biotype in extractions:
this_genes = extract_and_translate(items, gene_type, gene_biotype, transcript_type, transcript_biotype)
log.log("%d genes from %s/%s/%s/%s\n" % (len(this_genes), gene_type,gene_biotype,transcript_type,transcript_biotype))
genes.extend(this_genes)
log.log("\n")
genes = merge(genes,log)
features = [ ]
for gene in genes:
features.extend(all_within(gene))
features.sort(key=lambda i: (i.seqid,i.start))
return features
def get_genes(items, extractions, log):
annotation.link_up_annotations(items)
log.log("Content of downloaded GFF file (type:biotype ... paths):\n")
show_tree(items, log)
log.log("\n")
genes = list()
for gene_type,gene_biotype,transcript_type,transcript_biotype in extractions:
this_genes = extract_and_translate(items, gene_type, gene_biotype, transcript_type, transcript_biotype)
log.log("%d genes from %s/%s/%s/%s\n" % (len(this_genes), gene_type,gene_biotype,transcript_type,transcript_biotype))
genes.extend(this_genes)
log.log("\n")
genes = dominate(genes,log)
log.log("\n")
genes = merge(genes,log)
features = [ ]
for gene in genes:
features.extend(all_within(gene))
features.sort(key=lambda i: (i.seqid,i.start))
return features
def run(self):
workspace = self.get_workspace()
header = [ "##gff-version 3\n" ]
lengths = { }
with io.open_possibly_compressed_file(self.features) as f:
f.next()
for line in f:
if not line.startswith("#"): break
if line.startswith("##gff-version"): continue
header.append(line)
parts = line.strip().split()
if parts[0] == "##sequence-region":
lengths[parts[1]] = int(parts[3])
header = "".join(header)
items = list(annotation.read_gff(self.features, "/"))
annotation.link_up_annotations(items)
for item in items:
assert len(item.parents) < 2
if "ID" in item.attr:
item.attr["ID"] = item.attr["ID"].split(":")[1]
if "Parent" in item.attr:
item.attr["Parent"] = item.attr["Parent"].split(":")[1]
if item.parents:
item.parent = item.parents[0]
def well_supported(item):
if self.support is None: return True
level = item.attr.get("transcript_support_level","NA").split()[0]
if not level.isdigit(): return False
return int(level) <= self.support
exons = [ item for item in items if item.type == "exon" and well_supported(item.parent) ]
exon_index = span_index.index_annotations(exons)
utrs = [ ]
extended_utrs = [ ]
utr_parts = [ ]
exons_kept = [ ]
cds_kept = [ ]
transcripts_kept = [ ]
for item in items:
this_exons = [ item2 for item2 in item.children if item2.type == "exon" ]
if this_exons and well_supported(item):
transcripts_kept.append(item)
exons_kept.extend(this_exons)
cds_kept.extend([ item2 for item2 in item.children if item2.type == "CDS" ])
if self.gene_level:
utr_bits = [ item3 for item2 in item.children if well_supported(item2)
for item3 in item2.children if item3.type == self.what ]
else:
if not well_supported(item): continue
utr_bits = [ item2 for item2 in item.children if item2.type == self.what ]
if not utr_bits:
continue
utr = utr_bits[0].copy()
for item2 in utr_bits[1:]:
utr = utr.span_with(item2)
gene = item if self.gene_level else item.parent
utr.attr = dict(
ID=item.get_id(),
Name=item.attr["Name"],
gene_id=gene.get_id(),
gene=gene.attr["Name"],
description=gene.attr.get("description",""),
biotype=item.attr["biotype"]
)
max_extension = 10000
if item.strand < 0:
max_extension = min(max_extension, utr.start)
else:
max_extension = min(max_extension, lengths[utr.seqid] - utr.end)
assert max_extension >= 0, utr
end = utr.three_prime()
for hit in exon_index.get(end.shifted(0,max_extension), same_strand=True):
#if hit.parent.get_id() == item.get_id():
# continue
rel = hit.relative_to(end).start
if rel >= 0:
max_extension = min(max_extension, rel)
extended_utr = utr.shifted(0,max_extension)
extended_utr.start = max(extended_utr.start, 0)
utr.attr["max_extension"] = str(max_extension)
utrs.append(utr)
extended_utrs.append(extended_utr)
for item2 in utr_bits:
part = item2.copy()
part.attr = dict(Parent=item.get_id())
part.type = "part"
utr_parts.append(part)
write_gff3(workspace/"utr.gff",utrs,header)
write_gff3(workspace/"utr_extended.gff",extended_utrs,header)
write_gff3(workspace/"utr_part.gff",utr_parts,header)
write_gff3(workspace/"transcript.gff",transcripts_kept,header)
write_gff3(workspace/"exon.gff",exons_kept,header)
write_gff3(workspace/"cds.gff",cds_kept,header)
def run(self):
assert self.extension is not None, '--extension must be specified'
#workspace = self.get_workspace()
workspace = working_directory.Working(self.working_dir, must_exist=True)
if self.annotations == None:
reference = workspace.get_reference()
annotations_filename = reference.annotations_filename()
else:
annotations_filename = self.annotations
types = [ item.lower() for item in self.types.split(',') ]
parts = self.parts or self.types
parts = [ item.lower() for item in parts.split(',') ]
all_annotations = list(annotation.read_annotations(annotations_filename))
annotation.link_up_annotations(all_annotations)
for item in all_annotations:
item.primary = None
annotations = [
item
for item in all_annotations
if item.type.lower() in types
]
part_annotations = [ ]
seen = set()
queue = [ (item,item) for item in annotations ]
while queue:
primary, item = queue.pop()
if item.type.lower() in parts:
assert item.primary is None, "Feature with multiple parents"
item.primary = primary
key = (id(primary),item.start,item.end,item.seqid,item.strand)
# Ignore duplicate exons (many isoforms will have the same exons)
if key not in seen:
seen.add(key)
part_annotations.append(item)
queue.extend( (primary, item2) for item2 in item.children )
del seen
del all_annotations
self.log.log('%d annotations\n' % len(annotations))
self.log.log('%d part annotations\n' % len(part_annotations))
#assert annotations, 'No annotations of specified types in file'
for item in part_annotations:
this_extension = self.extension
if "max_extension" in item.attr:
this_extension = min(this_extension,int(item.attr["max_extension"]))
if item.strand >= 0:
item.tail_pos = item.end
item.end += this_extension
else:
item.tail_pos = item.start
item.start -= this_extension
for item in annotations:
item.hits = [] # [ (tail_length, adaptor_bases) ]
index = span_index.index_annotations(part_annotations)
for alignment in sam.Bam_reader(workspace/'alignments_filtered_sorted.bam'):
if alignment.is_unmapped or alignment.is_secondary or alignment.is_supplementary:
continue
start = alignment.reference_start
end = alignment.reference_end
alignment_length = end-start
strand = -1 if alignment.flag&sam.FLAG_REVERSE else 1
fragment_feature = annotation.Annotation(
seqid=alignment.reference_name,
start=start,
end=end,
strand=strand
)
if strand >= 0:
tail_pos = end
else:
tail_pos = start
tail_length = 0
adaptor_bases = 0
for item in alignment.extra:
if item.startswith('AN:i:'):
tail_length = int(item[5:])
elif item.startswith('AD:i:'):
adaptor_bases = int(item[5:])
hits = index.get(fragment_feature, same_strand=True)
if hits:
gene = min(hits, key=lambda gene:
(abs(tail_pos - gene.tail_pos), gene.primary.get_id()))
# Nearest by tail_pos
# failing that, by id to ensure a deterministic choice
gene.primary.hits.append( (tail_length,adaptor_bases) )
for item in annotations:
del item.parents
del item.children
del item.primary
f = io.open_possibly_compressed_writer(self.prefix + '.pickle.gz')
pickle.dump((workspace.name, workspace.get_tags(), annotations), f, pickle.HIGHEST_PROTOCOL)
f.close()
def run(self):
items = list(annotation.read_annotations(self.parent))
annotation.link_up_annotations(items)
for item in items:
assert len(item.parents) <= 1
genes = [ item for item in items if item.type == "gene" ]
downstrand_genes = [ _extend(_three_prime(item), self.extension) for item in genes ]
exons = [ item for item in items if item.type == "exon" ]
utrs = [ _extend(item, self.extension) for item in items if item.type == "three_prime_utr" ]
gene_index = span_index.index_annotations(genes)
downstrand_gene_index = span_index.index_annotations(downstrand_genes)
exon_index = span_index.index_annotations(exons)
utr_index = span_index.index_annotations(utrs)
peaks = list(annotation.read_annotations(self.child))
for peak in peaks:
# Query is final base in genome before poly(A) starts
query = peak.three_prime().shifted(-1,0)
hit_to = "3'UTR"
hits = [ item.parents[0].parents[0] for item in
utr_index.get(query, True) ]
if not hits:
hit_to = "Exon"
hits = [ item.parents[0].parents[0] for item in
exon_index.get(query, True) ]
# For non-coding RNAs, which don't have a 3' UTR
if not hits:
hit_to = "Downstrand"
hits = downstrand_gene_index.get(query, True)
if not hits:
hit_to = "Intron"
hits = gene_index.get(query, True)
antisense_hits = gene_index.get(query.reversed(), True)
if not hits:
hit_to = "Antisense"
hits = antisense_hits
if hits:
peak.attr["Parent"] = join_descriptions([ item.get_id() for item in hits ], ",")
peak.attr["Relation"] = hit_to
peak.attr["Name"] = join_descriptions([ item.attr.get("Name","") for item in hits ])
peak.attr["Product"] = hit_to + " " + join_descriptions([ item.attr.get("Product","") for item in hits ])
peak.attr["Biotype"] = join_descriptions([ item.attr.get("Biotype","") for item in hits ])
if antisense_hits:
peak.attr["Antisense_parent"] = join_descriptions([ item.get_id() for item in antisense_hits ], ",")
peak.attr["Antisense_name"] = join_descriptions([ item.attr.get("Name","") for item in antisense_hits ])
peak.attr["Antisense_product"] = "Antisense " + join_descriptions([ item.attr.get("Product","") for item in antisense_hits ])
peak.attr["Antisense_biotype"] = join_descriptions([ item.attr.get("Biotype","") for item in antisense_hits])
annotation.write_gff3(self.prefix+"-parent.gff", genes) #Hmm
annotation.write_gff3(self.prefix+"-child.gff", peaks)
def run(self):
work = self.get_workspace()
work.update_param(remove=['tail_tools_reference_version'])
nesoni.Make_reference(
self.output_dir,
filenames = self.filenames,
snpeff = False,
cs = 'ifavailable' if self.index else False,
ls = False,
bowtie = 'ifavailable' if self.index else False,
).run()
annotations = list(annotation.read_annotations(work/'reference.gff'))
annotation.link_up_annotations(annotations)
exon_index = span_index.index_annotations([
item for item in annotations if item.type == "exon"
])
mrna_end_index = span_index.index_annotations([
item.three_prime() for item in annotations if item.type == "mRNA"
])
mrna_utrs = [ ]
gene_utrs = [ ]
for gene in annotations:
if gene.type != 'gene': continue
mrnas = [ item for item in gene.children if item.type == 'mRNA' ]
assert mrnas, "Gene without any mRNAs: "+gene.get_id()
gene.attr['color'] = '#880088'
gene.start = min(item.start for item in mrnas)
gene.end = max(item.end for item in mrnas)
gene.attr["max_extension"] = str(_max_extension(gene, exon_index, mrna_end_index))
gene_utr_5primes = [ ]
for mrna in mrnas:
assert mrna.strand == gene.strand, mrna
assert mrna.seqid == gene.seqid, mrna
mrna.attr["max_extension"] = str(_max_extension(mrna, exon_index, mrna_end_index))
cdss = [ item for item in mrna.children if item.type == 'CDS' ]
exons = [ item for item in mrna.children if item.type == 'exon' ]
if not exons: continue
#link up annotations sorts children, so final is really final
for item in exons[:-1]:
item.attr["max_extension"] = "0"
exons[-1].attr["max_extension"] = mrna.attr["max_extension"]
if not cdss: continue
mrna_utr_5primes = [ ]
if gene.strand >= 0:
cds_3prime = max(item.end for item in cdss)
for item in exons:
if item.end >= cds_3prime:
mrna_utr_5primes.append(max(item.start,cds_3prime))
else:
cds_3prime = min(item.start for item in cdss)
for item in exons:
if item.start <= cds_3prime:
mrna_utr_5primes.append(min(item.end,cds_3prime))
if mrna.strand >= 0:
utr_start = min(mrna_utr_5primes) if mrna_utr_5primes else mrna.end
utr_end = max(utr_start+1,mrna.end)
gene_utr_5primes.append(utr_start)
else:
utr_end = max(mrna_utr_5primes) if mrna_utr_5primes else mrna.start
utr_start = min(mrna.start,utr_end-1)
gene_utr_5primes.append(utr_end)
attr = mrna.attr.copy()
attr['Parent'] = attr['ID']
attr['ID'] = attr['ID']+'-3UTR'
attr['color'] = '#008888'
utr = annotation.Annotation(
source = 'tt',
type = 'three_prime_utr',
seqid = mrna.seqid,
strand = mrna.strand,
start = utr_start,
end = utr_end,
attr = attr,
)
max_ext = _max_extension(utr, exon_index, mrna_end_index)
utr.attr["max_extension"] = str(max_ext)
#Only include if there is an annotated 3' UTR or end is not in the middle of some other isoform's exon
if utr_end-utr_start+max_ext > 1:
mrna_utrs.append(utr)
if gene.strand >= 0:
utr_start = max(gene_utr_5primes) if gene_utr_5primes else gene.end
utr_end = max(utr_start+1,gene.end)
else:
utr_end = min(gene_utr_5primes) if gene_utr_5primes else gene.start
utr_start = min(gene.start,utr_end-1)
attr = gene.attr.copy()
attr['Parent'] = attr['ID']
attr['ID'] = attr['ID']+'-3UTR'
attr['color'] = '#008888'
utr = annotation.Annotation(
source = 'tt',
type = 'three_prime_utr',
seqid = gene.seqid,
strand = gene.strand,
start = utr_start,
end = utr_end,
attr = attr,
)
utr.attr["max_extension"] = str(_max_extension(utr, exon_index, mrna_end_index))
gene_utrs.append(utr)
annotation.write_gff3(work/'reference.gff', annotations + mrna_utrs)
annotation.write_gff3(work/'utr.gff', gene_utrs)
work.update_param(tail_tools_reference_version=work.VERSION)
def run(self):
work = self.get_workspace()
work.update_param(remove=['tail_tools_reference_version'])
nesoni.Make_reference(
self.output_dir,
filenames = self.filenames,
snpeff = False,
cs = 'ifavailable' if self.index else False,
ls = False,
bowtie = 'ifavailable' if self.index else False,
).run()
annotations = list(annotation.read_annotations(work/'reference.gff'))
annotation.link_up_annotations(annotations)
with open(work/'utr.gff','wb') as f:
annotation.write_gff3_header(f)
for gene in annotations:
if gene.type != 'gene': continue
mrnas = [ item for item in gene.children if item.type == 'mRNA' ]
utr_5primes = [ ]
for mrna in mrnas:
cdss = [ item for item in mrna.children if item.type == 'CDS' ]
exons = [ item for item in mrna.children if item.type == 'exon' ]
if not cdss or not exons: continue
if gene.strand >= 0:
cds_3prime = max(item.end for item in cdss)
for item in exons:
if item.end > cds_3prime:
utr_5primes.append(max(item.start,cds_3prime))
else:
cds_3prime = min(item.start for item in cdss)
for item in exons:
if item.start < cds_3prime:
utr_5primes.append(min(item.end,cds_3prime))
if gene.strand >= 0:
utr_start = max(utr_5primes) if utr_5primes else gene.end
utr_end = max(utr_start+1,gene.end)
else:
utr_end = min(utr_5primes) if utr_5primes else gene.start
utr_start = min(gene.start,utr_end-1)
attr = gene.attr.copy()
attr['Parent'] = attr['ID']
attr['ID'] = attr['ID']+'-3UTR'
thing = annotation.Annotation(
source = 'tt',
type = 'three_prime_utr',
seqid = gene.seqid,
strand = gene.strand,
start = utr_start,
end = utr_end,
attr = attr,
)
print >> f, thing.as_gff()
work.update_param(tail_tools_reference_version=work.VERSION)
def run(self):
workspace = self.get_workspace()
header = ["##gff-version 3\n"]
lengths = {}
with io.open_possibly_compressed_file(self.features) as f:
f.next()
for line in f:
if not line.startswith("#"): break
if line.startswith("##gff-version"): continue
header.append(line)
parts = line.strip().split()
if parts[0] == "##sequence-region":
lengths[parts[1]] = int(parts[3])
header = "".join(header)
items = list(annotation.read_gff(self.features, "/"))
annotation.link_up_annotations(items)
for item in items:
assert len(item.parents) < 2
if "ID" in item.attr:
item.attr["ID"] = item.attr["ID"].split(":")[1]
if "Parent" in item.attr:
item.attr["Parent"] = item.attr["Parent"].split(":")[1]
if item.parents:
item.parent = item.parents[0]
def well_supported(item):
if self.support is None: return True
level = item.attr.get("transcript_support_level", "NA").split()[0]
if not level.isdigit(): return False
return int(level) <= self.support
exons = [
item for item in items
if item.type == "exon" and well_supported(item.parent)
]
exon_index = span_index.index_annotations(exons)
utrs = []
extended_utrs = []
utr_parts = []
exons_kept = []
cds_kept = []
transcripts_kept = []
for item in items:
this_exons = [
item2 for item2 in item.children if item2.type == "exon"
]
if this_exons and well_supported(item):
transcripts_kept.append(item)
exons_kept.extend(this_exons)
cds_kept.extend(
[item2 for item2 in item.children if item2.type == "CDS"])
if self.gene_level:
utr_bits = [
item3 for item2 in item.children if well_supported(item2)
for item3 in item2.children if item3.type == self.what
]
else:
if not well_supported(item): continue
utr_bits = [
item2 for item2 in item.children if item2.type == self.what
]
if not utr_bits:
continue
utr = utr_bits[0].copy()
for item2 in utr_bits[1:]:
utr = utr.span_with(item2)
gene = item if self.gene_level else item.parent
utr.attr = dict(ID=item.get_id(),
Name=item.attr["Name"],
gene_id=gene.get_id(),
gene=gene.attr["Name"],
description=gene.attr.get("description", ""),
biotype=item.attr["biotype"])
max_extension = 10000
if item.strand < 0:
max_extension = min(max_extension, utr.start)
else:
max_extension = min(max_extension,
lengths[utr.seqid] - utr.end)
assert max_extension >= 0, utr
end = utr.three_prime()
for hit in exon_index.get(end.shifted(0, max_extension),
same_strand=True):
#if hit.parent.get_id() == item.get_id():
# continue
rel = hit.relative_to(end).start
if rel >= 0:
max_extension = min(max_extension, rel)
extended_utr = utr.shifted(0, max_extension)
extended_utr.start = max(extended_utr.start, 0)
utr.attr["max_extension"] = str(max_extension)
utrs.append(utr)
extended_utrs.append(extended_utr)
for item2 in utr_bits:
part = item2.copy()
part.attr = dict(Parent=item.get_id())
part.type = "part"
utr_parts.append(part)
write_gff3(workspace / "utr.gff", utrs, header)
write_gff3(workspace / "utr_extended.gff", extended_utrs, header)
write_gff3(workspace / "utr_part.gff", utr_parts, header)
write_gff3(workspace / "transcript.gff", transcripts_kept, header)
write_gff3(workspace / "exon.gff", exons_kept, header)
write_gff3(workspace / "cds.gff", cds_kept, header)
def run(self):
work = self.get_workspace()
work.update_param(remove=['tail_tools_reference_version'])
nesoni.Make_reference(
self.output_dir,
filenames=self.filenames,
snpeff=False,
cs='ifavailable' if self.index else False,
ls=False,
bowtie='ifavailable' if self.index else False,
).run()
annotations = list(annotation.read_annotations(work / 'reference.gff'))
annotation.link_up_annotations(annotations)
exon_index = span_index.index_annotations(
[item for item in annotations if item.type == "exon"])
mrna_end_index = span_index.index_annotations([
item.three_prime() for item in annotations if item.type == "mRNA"
])
mrna_utrs = []
gene_utrs = []
for gene in annotations:
if gene.type != 'gene': continue
mrnas = [item for item in gene.children if item.type == 'mRNA']
assert mrnas, "Gene without any mRNAs: " + gene.get_id()
gene.attr['color'] = '#880088'
gene.start = min(item.start for item in mrnas)
gene.end = max(item.end for item in mrnas)
gene.attr["max_extension"] = str(
_max_extension(gene, exon_index, mrna_end_index))
gene_utr_5primes = []
for mrna in mrnas:
assert mrna.strand == gene.strand, mrna
assert mrna.seqid == gene.seqid, mrna
mrna.attr["max_extension"] = str(
_max_extension(mrna, exon_index, mrna_end_index))
cdss = [item for item in mrna.children if item.type == 'CDS']
exons = [item for item in mrna.children if item.type == 'exon']
if not exons: continue
#link up annotations sorts children, so final is really final
for item in exons[:-1]:
item.attr["max_extension"] = "0"
exons[-1].attr["max_extension"] = mrna.attr["max_extension"]
if not cdss: continue
mrna_utr_5primes = []
if gene.strand >= 0:
cds_3prime = max(item.end for item in cdss)
for item in exons:
if item.end >= cds_3prime:
mrna_utr_5primes.append(max(
item.start, cds_3prime))
else:
cds_3prime = min(item.start for item in cdss)
for item in exons:
if item.start <= cds_3prime:
mrna_utr_5primes.append(min(item.end, cds_3prime))
if mrna.strand >= 0:
utr_start = min(
mrna_utr_5primes) if mrna_utr_5primes else mrna.end
utr_end = max(utr_start + 1, mrna.end)
gene_utr_5primes.append(utr_start)
else:
utr_end = max(
mrna_utr_5primes) if mrna_utr_5primes else mrna.start
utr_start = min(mrna.start, utr_end - 1)
gene_utr_5primes.append(utr_end)
attr = mrna.attr.copy()
attr['Parent'] = attr['ID']
attr['ID'] = attr['ID'] + '-3UTR'
attr['color'] = '#008888'
utr = annotation.Annotation(
source='tt',
type='three_prime_utr',
seqid=mrna.seqid,
strand=mrna.strand,
start=utr_start,
end=utr_end,
attr=attr,
)
max_ext = _max_extension(utr, exon_index, mrna_end_index)
utr.attr["max_extension"] = str(max_ext)
#Only include if there is an annotated 3' UTR or end is not in the middle of some other isoform's exon
if utr_end - utr_start + max_ext > 1:
mrna_utrs.append(utr)
if gene.strand >= 0:
utr_start = max(
gene_utr_5primes) if gene_utr_5primes else gene.end
utr_end = max(utr_start + 1, gene.end)
else:
utr_end = min(
gene_utr_5primes) if gene_utr_5primes else gene.start
utr_start = min(gene.start, utr_end - 1)
attr = gene.attr.copy()
attr['Parent'] = attr['ID']
attr['ID'] = attr['ID'] + '-3UTR'
attr['color'] = '#008888'
utr = annotation.Annotation(
source='tt',
type='three_prime_utr',
seqid=gene.seqid,
strand=gene.strand,
start=utr_start,
end=utr_end,
attr=attr,
)
utr.attr["max_extension"] = str(
_max_extension(utr, exon_index, mrna_end_index))
gene_utrs.append(utr)
annotation.write_gff3(work / 'reference.gff', annotations + mrna_utrs)
annotation.write_gff3(work / 'utr.gff', gene_utrs)
work.update_param(tail_tools_reference_version=work.VERSION)
def run(self):
items = list(annotation.read_annotations(self.parent))
annotation.link_up_annotations(items)
for item in items:
assert len(item.parents) <= 1
genes = [ item for item in items if item.type == "gene" ]
downstrand_genes = [ _extend(_three_prime(item), self.extension) for item in genes ]
exons = [ item for item in items if item.type == "exon" ]
utrs = [ _extend(item, self.extension) for item in items if item.type == "three_prime_utr" ]
gene_index = span_index.index_annotations(genes)
downstrand_gene_index = span_index.index_annotations(downstrand_genes)
exon_index = span_index.index_annotations(exons)
utr_index = span_index.index_annotations(utrs)
peaks = [ ]
for peak in annotation.read_annotations(self.child):
if float(peak.attr.get("mean_tail","0.0")) < self.min_tail:
continue
peaks.append(peak)
for peak in peaks:
# Query is final base in genome before poly(A) starts
query = peak.three_prime().shifted(-1,0)
hit_to = "3'UTR"
hits = [ item.parents[0].parents[0] for item in
utr_index.get(query, True) ]
if not hits:
hit_to = "Exon"
hits = [ item.parents[0].parents[0] for item in
exon_index.get(query, True) ]
# For non-coding RNAs, which don't have a 3' UTR
if not hits:
hit_to = "Downstrand"
hits = downstrand_gene_index.get(query, True)
if not hits:
hit_to = "Intron"
hits = gene_index.get(query, True)
antisense_hits = gene_index.get(query.reversed(), True)
if not hits:
hit_to = "Antisense"
hits = antisense_hits
if hits:
peak.attr["Parent"] = join_descriptions([ item.get_id() for item in hits ], ",")
peak.attr["Relation"] = hit_to
peak.attr["Name"] = join_descriptions([ item.attr.get("Name","") for item in hits ])
peak.attr["Product"] = hit_to + " " + join_descriptions([ item.attr.get("Product","") for item in hits ])
peak.attr["Biotype"] = join_descriptions([ item.attr.get("Biotype","") for item in hits ])
if antisense_hits:
peak.attr["Antisense_parent"] = join_descriptions([ item.get_id() for item in antisense_hits ], ",")
peak.attr["Antisense_name"] = join_descriptions([ item.attr.get("Name","") for item in antisense_hits ])
peak.attr["Antisense_product"] = "Antisense " + join_descriptions([ item.attr.get("Product","") for item in antisense_hits ])
peak.attr["Antisense_biotype"] = join_descriptions([ item.attr.get("Biotype","") for item in antisense_hits])
annotation.write_gff3(self.prefix+"-parent.gff", genes) #Hmm
annotation.write_gff3(self.prefix+"-child.gff", peaks)
def run(self):
assert self.extension is not None, '--extension must be specified'
#workspace = self.get_workspace()
workspace = working_directory.Working(self.working_dir,
must_exist=True)
if self.annotations == None:
reference = workspace.get_reference()
annotations_filename = reference.annotations_filename()
else:
annotations_filename = self.annotations
types = [item.lower() for item in self.types.split(',')]
parts = self.parts or self.types
parts = [item.lower() for item in parts.split(',')]
all_annotations = list(
annotation.read_annotations(annotations_filename))
annotation.link_up_annotations(all_annotations)
for item in all_annotations:
item.primary = None
annotations = [
item for item in all_annotations if item.type.lower() in types
]
part_annotations = []
seen = set()
queue = [(item, item) for item in annotations]
while queue:
primary, item = queue.pop()
if item.type.lower() in parts:
assert item.primary is None, "Feature with multiple parents"
item.primary = primary
key = (id(primary), item.start, item.end, item.seqid,
item.strand)
# Ignore duplicate exons (many isoforms will have the same exons)
if key not in seen:
seen.add(key)
part_annotations.append(item)
queue.extend((primary, item2) for item2 in item.children)
del seen
del all_annotations
self.log.log('%d annotations\n' % len(annotations))
self.log.log('%d part annotations\n' % len(part_annotations))
#assert annotations, 'No annotations of specified types in file'
for item in part_annotations:
this_extension = self.extension
if "max_extension" in item.attr:
this_extension = min(this_extension,
int(item.attr["max_extension"]))
if item.strand >= 0:
item.tail_pos = item.end
item.end += this_extension
else:
item.tail_pos = item.start
item.start -= this_extension
for item in annotations:
item.hits = [] # [ (tail_length, adaptor_bases) ]
index = span_index.index_annotations(part_annotations)
for alignment in sam.Bam_reader(workspace /
'alignments_filtered_sorted.bam'):
if alignment.is_unmapped or alignment.is_secondary or alignment.is_supplementary:
continue
start = alignment.reference_start
end = alignment.reference_end
alignment_length = end - start
strand = -1 if alignment.flag & sam.FLAG_REVERSE else 1
fragment_feature = annotation.Annotation(
seqid=alignment.reference_name,
start=start,
end=end,
strand=strand)
if strand >= 0:
tail_pos = end
else:
tail_pos = start
tail_length = 0
adaptor_bases = 0
for item in alignment.extra:
if item.startswith('AN:i:'):
tail_length = int(item[5:])
elif item.startswith('AD:i:'):
adaptor_bases = int(item[5:])
hits = index.get(fragment_feature, same_strand=True)
if hits:
gene = min(
hits,
key=lambda gene:
(abs(tail_pos - gene.tail_pos), gene.primary.get_id()))
# Nearest by tail_pos
# failing that, by id to ensure a deterministic choice
gene.primary.hits.append((tail_length, adaptor_bases))
for item in annotations:
del item.parents
del item.children
del item.primary
f = io.open_possibly_compressed_writer(self.prefix + '.pickle.gz')
pickle.dump((workspace.name, workspace.get_tags(), annotations), f,
pickle.HIGHEST_PROTOCOL)
f.close()