#!/usr/bin/env python """ usage: %prog $input $out_file1 -1, --cols=N,N,N,N,N: Columns for start, end, strand in input file -d, --dbkey=N: Genome build of input file -o, --output_format=N: the data type of the output file -g, --GALAXY_DATA_INDEX_DIR=N: the directory containing alignseq.loc or twobit.loc -I, --interpret_features: if true, complete features are interpreted when input is GFF -F, --fasta=: genomic sequences to use for extraction -G, --gff: input and output file, when it is interval, coordinates are treated as GFF format (1-based, half-open) rather than 'traditional' 0-based, closed format. """ from __future__ import print_function import os import subprocess import sys import tempfile import bx.seq.nib import bx.seq.twobit from bx.cookbook import doc_optparse from bx.tabular.io import ( Comment, Header, ) from galaxy.datatypes.util import gff_util from galaxy.tools.util.galaxyops import parse_cols_arg def stop_err(msg): sys.stderr.write(msg) sys.exit() def reverse_complement(s): complement_dna = { "A": "T", "T": "A", "C": "G", "G": "C", "a": "t", "t": "a", "c": "g", "g": "c", "N": "N", "n": "n", } reversed_s = [] for i in s: reversed_s.append(complement_dna[i]) reversed_s.reverse() return "".join(reversed_s) def check_seq_file(dbkey, GALAXY_DATA_INDEX_DIR): # Checks for the presence of *.nib files matching the dbkey within alignseq.loc seq_file = "%s/alignseq.loc" % GALAXY_DATA_INDEX_DIR for line in open(seq_file): line = line.rstrip("\r\n") if line and not line.startswith("#") and line.startswith("seq"): fields = line.split("\t") if len(fields) >= 3 and fields[1] == dbkey: print("Using *.nib genomic reference files") return fields[2].strip() # If no entry in aligseq.loc was found, check for the presence of a *.2bit file in twobit.loc seq_file = "%s/twobit.loc" % GALAXY_DATA_INDEX_DIR for line in open(seq_file): line = line.rstrip("\r\n") if line and not line.startswith("#") and line.endswith(".2bit"): fields = line.split("\t") if len(fields) >= 2 and fields[0] == dbkey: print("Using a *.2bit genomic reference file") return fields[1].strip() return "" def __main__(): # # Parse options, args. # options, args = doc_optparse.parse(__doc__) try: if len(options.cols.split(",")) == 5: # BED file chrom_col, start_col, end_col, strand_col, name_col = parse_cols_arg(options.cols) else: # gff file chrom_col, start_col, end_col, strand_col = parse_cols_arg(options.cols) name_col = False dbkey = options.dbkey output_format = options.output_format gff_format = options.gff interpret_features = options.interpret_features GALAXY_DATA_INDEX_DIR = options.GALAXY_DATA_INDEX_DIR fasta_file = options.fasta input_filename, output_filename = args except Exception: doc_optparse.exception() includes_strand_col = strand_col >= 0 strand = None nibs = {} # # Set path to sequence data. # if fasta_file: # Need to create 2bit file from fasta file. try: seq_path = tempfile.NamedTemporaryFile(dir=".").name cmd = "faToTwoBit %s %s" % (fasta_file, seq_path) tmp_name = tempfile.NamedTemporaryFile(dir=".").name tmp_stderr = open(tmp_name, "wb") proc = subprocess.Popen(args=cmd, shell=True, stderr=tmp_stderr.fileno()) returncode = proc.wait() tmp_stderr.close() # Get stderr, allowing for case where it's very large. tmp_stderr = open(tmp_name, "rb") stderr = "" buffsize = 1048576 try: while True: stderr += tmp_stderr.read(buffsize) if not stderr or len(stderr) % buffsize != 0: break except OverflowError: pass tmp_stderr.close() # Error checking. if returncode != 0: raise Exception(stderr) except Exception as e: stop_err("Error running faToTwoBit. " + str(e)) else: seq_path = check_seq_file(dbkey, GALAXY_DATA_INDEX_DIR) if not os.path.exists(seq_path): # If this occurs, we need to fix the metadata validator. stop_err("No sequences are available for '%s', request them by reporting this error." % dbkey) # # Fetch sequences. # # Get feature's line(s). def get_lines(feature): if isinstance(feature, gff_util.GFFFeature): return feature.lines() else: return [feature.rstrip("\r\n")] skipped_lines = 0 first_invalid_line = 0 invalid_lines = [] fout = open(output_filename, "w") warnings = [] warning = "" twobitfile = None file_iterator = open(input_filename) if gff_format and interpret_features: file_iterator = gff_util.GFFReaderWrapper(file_iterator, fix_strand=False) line_count = 1 for feature in file_iterator: # Ignore comments, headers. if isinstance(feature, (Header, Comment)): line_count += 1 continue name = "" if gff_format and interpret_features: # Processing features. gff_util.convert_gff_coords_to_bed(feature) chrom = feature.chrom start = feature.start end = feature.end strand = feature.strand else: # Processing lines, either interval or GFF format. line = feature.rstrip("\r\n") if line and not line.startswith("#"): fields = line.split("\t") try: chrom = fields[chrom_col] start = int(fields[start_col]) end = int(fields[end_col]) if name_col: name = fields[name_col] if gff_format: start, end = gff_util.convert_gff_coords_to_bed([start, end]) if includes_strand_col: strand = fields[strand_col] except Exception: warning = "Invalid chrom, start or end column values. " warnings.append(warning) if not invalid_lines: invalid_lines = get_lines(feature) first_invalid_line = line_count skipped_lines += len(invalid_lines) continue if start > end: warning = "Invalid interval, start '%d' > end '%d'. " % (start, end) warnings.append(warning) if not invalid_lines: invalid_lines = get_lines(feature) first_invalid_line = line_count skipped_lines += len(invalid_lines) continue if strand not in ["+", "-"]: strand = "+" sequence = "" else: continue # Open sequence file and get sequence for feature/interval. if seq_path and os.path.exists("%s/%s.nib" % (seq_path, chrom)): # TODO: improve support for GFF-nib interaction. if chrom in nibs: nib = nibs[chrom] else: nibs[chrom] = nib = bx.seq.nib.NibFile(open("%s/%s.nib" % (seq_path, chrom))) try: sequence = nib.get(start, end - start) except Exception: warning = "Unable to fetch the sequence from '%d' to '%d' for build '%s'. " % ( start, end - start, dbkey, ) warnings.append(warning) if not invalid_lines: invalid_lines = get_lines(feature) first_invalid_line = line_count skipped_lines += len(invalid_lines) continue elif seq_path and os.path.isfile(seq_path): if not (twobitfile): twobitfile = bx.seq.twobit.TwoBitFile(open(seq_path, "rb")) try: if options.gff and interpret_features: # Create sequence from intervals within a feature. sequence = "" for interval in feature.intervals: sequence += twobitfile[interval.chrom][interval.start : interval.end] else: sequence = twobitfile[chrom][start:end] except Exception: warning = "Unable to fetch the sequence from '%d' to '%d' for chrom '%s'. " % ( start, end - start, chrom, ) warnings.append(warning) if not invalid_lines: invalid_lines = get_lines(feature) first_invalid_line = line_count skipped_lines += len(invalid_lines) continue else: warning = "Chromosome by name '%s' was not found for build '%s'. " % (chrom, dbkey) warnings.append(warning) if not invalid_lines: invalid_lines = get_lines(feature) first_invalid_line = line_count skipped_lines += len(invalid_lines) continue if sequence == "": warning = "Chrom: '%s', start: '%s', end: '%s' is either invalid or not present in build '%s'. " % ( chrom, start, end, dbkey, ) warnings.append(warning) if not invalid_lines: invalid_lines = get_lines(feature) first_invalid_line = line_count skipped_lines += len(invalid_lines) continue if includes_strand_col and strand == "-": sequence = reverse_complement(sequence) if output_format == "fasta": len_sequence = len(sequence) c = 0 if gff_format: start, end = gff_util.convert_bed_coords_to_gff([start, end]) fields = [dbkey, str(chrom), str(start), str(end), strand] meta_data = "_".join(fields) if name.strip(): fout.write(">%s %s\n" % (meta_data, name)) else: fout.write(">%s\n" % meta_data) while c < len_sequence: b = min(c + 50, len_sequence) fout.write("%s\n" % str(sequence[c:b])) c = b else: # output_format == "interval" if gff_format and interpret_features: # TODO: need better GFF Reader to capture all information needed # to produce this line. meta_data = "\t".join( [ feature.chrom, "galaxy_extract_genomic_dna", "interval", str(feature.start), str(feature.end), feature.score, feature.strand, ".", gff_util.gff_attributes_to_str(feature.attributes, "GTF"), ] ) else: meta_data = "\t".join(fields) if gff_format: format_str = '%s seq "%s";\n' else: format_str = "%s\t%s\n" fout.write(format_str % (meta_data, str(sequence))) # Update line count. if isinstance(feature, gff_util.GFFFeature): line_count += len(feature.intervals) else: line_count += 1 fout.close() if warnings: warn_msg = "%d warnings, 1st is: " % len(warnings) warn_msg += warnings[0] print(warn_msg) if skipped_lines: # Error message includes up to the first 10 skipped lines. print( 'Skipped %d invalid lines, 1st is #%d, "%s"' % (skipped_lines, first_invalid_line, "\n".join(invalid_lines[:10])) ) # Clean up temp file. if fasta_file: os.remove(seq_path) os.remove(tmp_name) if __name__ == "__main__": __main__()