Mercurial > repos > jjohnson > translate_bed_sequences
annotate translate_bed_sequences.py @ 3:3b526a780849
Change default seqtype from pep:novel to pep:splice
| author | Jim Johnson <jj@umn.edu> |
|---|---|
| date | Thu, 23 Jan 2014 09:10:16 -0600 |
| parents | 359addb9b9d4 |
| children | c626a939eef7 |
| rev | line source |
|---|---|
| 0 | 1 #!/usr/bin/env python |
| 2 """ | |
| 3 # | |
| 4 #------------------------------------------------------------------------------ | |
| 5 # University of Minnesota | |
| 6 # Copyright 2014, Regents of the University of Minnesota | |
| 7 #------------------------------------------------------------------------------ | |
| 8 # Author: | |
| 9 # | |
| 10 # James E Johnson | |
| 11 # | |
| 12 #------------------------------------------------------------------------------ | |
| 13 """ | |
| 14 | |
| 15 """ | |
| 16 Input: BED file (12 column) + 13th sequence column appended by extract_genomic_dna | |
| 17 Output: Fasta of 3-frame translations of the spliced sequence | |
| 18 | |
| 19 """ | |
| 20 | |
| 21 import sys,re,os.path | |
| 22 import optparse | |
| 23 from optparse import OptionParser | |
| 24 from Bio.Seq import reverse_complement, transcribe, back_transcribe, translate | |
| 25 | |
| 26 class BedEntry( object ): | |
| 27 def __init__(self, line): | |
| 28 self.line = line | |
| 29 try: | |
| 30 (chrom,chromStart,chromEnd,name,score,strand,thickStart,thickEnd,itemRgb,blockCount,blockSizes,blockStarts,seq) = line.split('\t')[0:13] | |
| 31 self.chrom = chrom | |
| 32 self.chromStart = int(chromStart) | |
| 33 self.chromEnd = int(chromEnd) | |
| 34 self.name = name | |
| 35 self.score = int(score) | |
| 36 self.strand = strand | |
| 37 self.thickStart = int(thickStart) | |
| 38 self.thickEnd = int(thickEnd) | |
| 39 self.itemRgb = itemRgb | |
| 40 self.blockCount = int(blockCount) | |
| 41 self.blockSizes = [int(x) for x in blockSizes.split(',')] | |
| 42 self.blockStarts = [int(x) for x in blockStarts.split(',')] | |
| 43 self.seq = seq | |
| 44 except Exception, e: | |
| 45 print >> sys.stderr, "Unable to read Bed entry" % e | |
| 46 exit(1) | |
| 47 def get_splice_junctions(self): | |
| 48 splice_juncs = [] | |
| 49 for i in range(self.blockCount - 1): | |
| 50 splice_junc = "%s:%d_%d" % (self.chrom, self.chromStart + self.blockSizes[i], self.chromStart + self.blockStarts[i+1]) | |
| 51 splice_juncs.append(splice_junc) | |
| 52 return splice_juncs | |
| 53 def get_exon_seqs(self): | |
| 54 exons = [] | |
| 55 for i in range(self.blockCount): | |
| 56 # splice_junc = "%s:%d_%d" % (self.chrom, self.chromStart + self.blockSizes[i], self.chromStart + self.blockStarts[i+1]) | |
| 57 exons.append(self.seq[self.blockStarts[i]:self.blockStarts[i] + self.blockSizes[i]]) | |
| 58 if self.strand == '-': #reverse complement | |
| 59 exons.reverse() | |
| 60 for i,s in enumerate(exons): | |
| 61 exons[i] = reverse_complement(s) | |
| 62 return exons | |
| 63 def get_spliced_seq(self): | |
| 64 seq = ''.join(self.get_exon_seqs()) | |
| 65 return seq | |
| 66 def get_translation(self,sequence=None): | |
| 67 translation = None | |
| 68 seq = sequence if sequence else self.get_spliced_seq() | |
| 69 if seq: | |
| 70 seqlen = len(seq) / 3 * 3; | |
| 71 if seqlen >= 3: | |
| 72 translation = translate(seq[:seqlen]) | |
| 73 return translation | |
| 74 def get_translations(self): | |
| 75 translations = [] | |
| 76 seq = self.get_spliced_seq() | |
| 77 if seq: | |
| 78 for i in range(3): | |
| 79 translation = self.get_translation(sequence=seq[i:]) | |
| 80 if translation: | |
| 81 translations.append(translation) | |
| 82 return translations | |
| 83 ## [[start,end,seq],[start,end,seq],[start,end,seq]] | |
| 84 ## filter: ignore translation if stop codon in first exon after ignore_left_bp | |
| 85 def get_filterd_translations(self,untrimmed=False,filtering=True,ignore_left_bp=0,ignore_right_bp=0): | |
| 86 translations = [None,None,None] | |
| 87 seq = self.get_spliced_seq() | |
| 88 ignore = (ignore_left_bp if self.strand == '+' else ignore_right_bp) / 3 | |
| 89 block_sum = sum(self.blockSizes) | |
| 90 exon_sizes = self.blockSizes | |
| 91 if self.strand == '-': | |
| 92 exon_sizes.reverse() | |
| 93 splice_sites = [sum(exon_sizes[:x]) / 3 for x in range(1,len(exon_sizes))] | |
| 94 junc = splice_sites[0] if len(splice_sites) > 0 else exon_sizes[0] | |
| 95 if seq: | |
| 96 for i in range(3): | |
| 97 translation = self.get_translation(sequence=seq[i:]) | |
| 98 if translation: | |
| 99 tstart = 0 | |
| 100 tstop = len(translation) | |
| 101 if not untrimmed: | |
| 102 tstart = translation.rfind('*',0,junc) + 1 | |
|
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359addb9b9d4
Do not include the stop codon when filtering
Jim Johnson <jj@umn.edu>
parents:
0
diff
changeset
|
103 stop = translation.find('*',junc) |
|
359addb9b9d4
Do not include the stop codon when filtering
Jim Johnson <jj@umn.edu>
parents:
0
diff
changeset
|
104 tstop = stop if stop >= 0 else len(translation) |
| 0 | 105 if filtering and tstart > ignore: |
| 106 continue | |
| 107 trimmed = translation[tstart:tstop] | |
| 108 #get genomic locations for start and end | |
| 109 offset = (block_sum - i) % 3 | |
| 110 if self.strand == '+': | |
| 111 chromStart = self.chromStart + i + (tstart * 3) | |
| 112 chromEnd = self.chromEnd - offset - (len(translation) - tstop) * 3 | |
| 113 else: | |
| 114 chromStart = self.chromStart + offset + (len(translation) - tstop) * 3 | |
| 115 chromEnd = self.chromEnd - i - (tstart * 3) | |
| 116 translations[i] = [chromStart,chromEnd,trimmed] | |
| 117 return translations | |
| 118 def get_seq_id(self,seqtype='unk:unk',reference='',frame=None): | |
| 119 ## Ensembl fasta ID format | |
| 120 # >ID SEQTYPE:STATUS LOCATION GENE TRANSCRIPT | |
|
3
3b526a780849
Change default seqtype from pep:novel to pep:splice
Jim Johnson <jj@umn.edu>
parents:
2
diff
changeset
|
121 # >ENSP00000328693 pep:splice chromosome:NCBI35:1:904515:910768:1 gene:ENSG00000158815:transcript:ENST00000328693 gene_biotype:protein_coding transcript_biotype:protein_coding |
| 0 | 122 frame_name = '' |
| 123 chromStart = self.chromStart | |
| 124 chromEnd = self.chromEnd | |
| 125 strand = 1 if self.strand == '+' else -1 | |
| 126 if frame != None: | |
| 127 block_sum = sum(self.blockSizes) | |
| 128 offset = (block_sum - frame) % 3 | |
| 129 frame_name = '_' + str(frame + 1) | |
| 130 if self.strand == '+': | |
| 131 chromStart += frame | |
| 132 chromEnd -= offset | |
| 133 else: | |
| 134 chromStart += offset | |
| 135 chromEnd -= frame | |
| 136 location = "chromosome:%s:%s:%s:%s:%s" % (reference,self.chrom,chromStart,chromEnd,strand) | |
| 137 seq_id = "%s%s %s %s" % (self.name,frame_name,seqtype,location) | |
| 138 return seq_id | |
| 139 def get_line(self, start_offset = 0, end_offset = 0): | |
| 140 if start_offset or end_offset: | |
| 141 s_offset = start_offset if start_offset else 0 | |
| 142 e_offset = end_offset if end_offset else 0 | |
| 143 if s_offset > self.chromStart: | |
| 144 s_offset = self.chromStart | |
| 145 chrStart = self.chromStart - s_offset | |
| 146 chrEnd = self.chromEnd + e_offset | |
| 147 blkSizes = self.blockSizes | |
| 148 blkSizes[0] += s_offset | |
| 149 blkSizes[-1] += e_offset | |
| 150 blkStarts = self.blockStarts | |
| 151 for i in range(1,self.blockCount): | |
| 152 blkStarts[i] += s_offset | |
| 153 items = [str(x) for x in [self.chrom,chrStart,chrEnd,self.name,self.score,self.strand,self.thickStart,self.thickEnd,self.itemRgb,self.blockCount,','.join([str(x) for x in blkSizes]),','.join([str(x) for x in blkStarts])]] | |
| 154 return '\t'.join(items) + '\n' | |
| 155 return self.line | |
| 156 | |
| 157 def __main__(): | |
| 158 #Parse Command Line | |
| 159 parser = optparse.OptionParser() | |
| 160 parser.add_option( '-i', '--input', dest='input', help='BED file (tophat junctions.bed) with sequence column added' ) | |
| 161 parser.add_option( '-o', '--output', dest='output', help='Translations of spliced sequence') | |
| 162 parser.add_option( '-b', '--bed_format', dest='bed_format', action='store_true', default=False, help='Append translations to bed file instead of fasta' ) | |
|
3
3b526a780849
Change default seqtype from pep:novel to pep:splice
Jim Johnson <jj@umn.edu>
parents:
2
diff
changeset
|
163 parser.add_option( '-S', '--seqtype', dest='seqtype', default='pep:splice', help='SEQTYPE:STATUS for fasta ID line' ) |
| 0 | 164 parser.add_option( '-R', '--reference', dest='reference', default=None, help='Genome Reference Name for fasta ID location ' ) |
| 165 parser.add_option( '-Q', '--score_name', dest='score_name', default=None, help='include in the fasta ID line score_name:score ' ) | |
| 166 parser.add_option( '-l', '--leading_bp', dest='leading_bp', type='int', default=None, help='leading number of base pairs to ignore when filtering' ) | |
| 167 parser.add_option( '-t', '--trailing_bp', dest='trailing_bp', type='int', default=None, help='trailing number of base pairs to ignore when filtering' ) | |
| 168 parser.add_option( '-U', '--unfiltered', dest='filtering', action='store_false', default=True, help='Do NOT filterout translation with stop codon in the first exon' ) | |
| 169 parser.add_option( '-u', '--untrimmed', dest='untrimmed', action='store_true', default=False, help='Do NOT trim from splice site to stop codon' ) | |
| 170 parser.add_option( '-L', '--min_length', dest='min_length', type='int', default=None, help='Minimun length (to first stop codon)' ) | |
| 171 parser.add_option( '-M', '--max_stop_codons', dest='max_stop_codons', type='int', default=None, help='Filter out translations with more than max_stop_codons' ) | |
| 172 parser.add_option( '-d', '--debug', dest='debug', action='store_true', default=False, help='Turn on wrapper debugging to stdout' ) | |
| 173 (options, args) = parser.parse_args() | |
| 174 # Input files | |
| 175 if options.input != None: | |
| 176 try: | |
| 177 inputPath = os.path.abspath(options.input) | |
| 178 inputFile = open(inputPath, 'r') | |
| 179 except Exception, e: | |
| 180 print >> sys.stderr, "failed: %s" % e | |
| 181 exit(2) | |
| 182 else: | |
| 183 inputFile = sys.stdin | |
| 184 # Output files | |
| 185 outFile = None | |
| 186 if options.output == None: | |
| 187 #write to stdout | |
| 188 outFile = sys.stdout | |
| 189 else: | |
| 190 try: | |
| 191 outPath = os.path.abspath(options.output) | |
| 192 outFile = open(outPath, 'w') | |
| 193 except Exception, e: | |
| 194 print >> sys.stderr, "failed: %s" % e | |
| 195 exit(3) | |
| 196 leading_bp = 0 | |
| 197 trailing_bp = 0 | |
| 198 if options.leading_bp: | |
| 199 if options.leading_bp >= 0: | |
| 200 leading_bp = options.leading_bp | |
| 201 else: | |
| 202 print >> sys.stderr, "failed: leading_bp must be positive" | |
| 203 exit(5) | |
| 204 if options.trailing_bp: | |
| 205 if options.trailing_bp >= 0: | |
| 206 trailing_bp = options.trailing_bp | |
| 207 else: | |
| 208 print >> sys.stderr, "failed: trailing_bp must be positive" | |
| 209 exit(5) | |
| 210 # Scan bed file | |
| 211 try: | |
| 212 for i, line in enumerate( inputFile ): | |
| 213 if line.startswith('track'): | |
| 214 if outFile and options.bed_format: | |
| 215 outFile.write(line) | |
| 216 continue | |
| 217 entry = BedEntry(line) | |
| 218 strand = 1 if entry.strand == '+' else -1 | |
| 219 translations = entry.get_translations() | |
| 220 if options.debug: | |
| 221 exon_seqs = entry.get_exon_seqs() | |
| 222 exon_sizes = [len(seq) for seq in exon_seqs] | |
| 223 splice_sites = [sum(exon_sizes[:x]) / 3 for x in range(1,len(exon_sizes))] | |
| 224 print >> sys.stderr, entry.name | |
| 225 print >> sys.stderr, line.rstrip('\r\n') | |
| 226 print >> sys.stderr, "exons: %s" % exon_seqs | |
| 227 print >> sys.stderr, "%s" % splice_sites | |
| 228 for i,translation in enumerate(translations): | |
| 229 print >> sys.stderr, "frame %d: %s" % (i+1,translation) | |
| 230 print >> sys.stderr, "splice: %s" % (''.join(['^' if (((j*3)+i)/3) in splice_sites else '-' for j in range(len(translation))])) | |
| 231 print >> sys.stderr, "" | |
| 232 if options.bed_format: | |
| 233 tx_entry = "%s\t%s\n" % (line.rstrip('\r\n'),'\t'.join(translations)) | |
| 234 outFile.write(tx_entry) | |
| 235 else: | |
| 236 translations = entry.get_filterd_translations(untrimmed=options.untrimmed,filtering=options.filtering,ignore_left_bp=leading_bp,ignore_right_bp=trailing_bp) | |
| 237 for i,tx in enumerate(translations): | |
| 238 if tx: | |
| 239 (chromStart,chromEnd,translation) = tx | |
| 240 if options.min_length != None and len(translation) < options.min_length: | |
| 241 continue | |
| 242 if options.max_stop_codons != None and translation.count('*') > options.max_stop_codons: | |
| 243 continue | |
| 244 frame_name = '_%s' % (i + 1) | |
| 245 location = "chromosome:%s:%s:%s:%s:%s" % (options.reference,entry.chrom,chromStart,chromEnd,strand) | |
| 246 score = " %s:%s" % (options.score_name,entry.score) if options.score_name else '' | |
| 247 seq_id = "%s%s %s %s%s" % (entry.name,frame_name,options.seqtype,location, score) | |
| 248 outFile.write(">%s\n" % seq_id) | |
| 249 outFile.write(translation) | |
| 250 outFile.write('\n') | |
| 251 except Exception, e: | |
| 252 print >> sys.stderr, "failed: Error reading %s - %s" % (options.input if options.input else 'stdin',e) | |
| 253 | |
| 254 if __name__ == "__main__" : __main__() | |
| 255 |