annotate change_o/MakeDb.py @ 65:dc06e94bc1e3 draft

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author davidvanzessen
date Wed, 06 Dec 2017 09:55:37 -0500
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1 #!/usr/bin/env python3
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2 """
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3 Create tab-delimited database file to store sequence alignment information
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4 """
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5 # Info
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6 __author__ = 'Namita Gupta, Jason Anthony Vander Heiden'
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7 from changeo import __version__, __date__
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8
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9 # Imports
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10 import os
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11 import sys
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12 from argparse import ArgumentParser
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13 from collections import OrderedDict
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14 from textwrap import dedent
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15 from time import time
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16 from Bio import SeqIO
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17
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18 # Presto and changeo imports
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19 from presto.Defaults import default_out_args
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20 from presto.Annotation import parseAnnotation
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21 from presto.IO import countSeqFile, printLog, printMessage, printProgress, readSeqFile
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22 from changeo.Commandline import CommonHelpFormatter, getCommonArgParser, parseCommonArgs
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23 from changeo.IO import countDbFile, extractIMGT, getDbWriter, readRepo
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24 from changeo.Parsers import IgBLASTReader, IMGTReader, IHMMuneReader, getIDforIMGT
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26
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27 def getFilePrefix(aligner_output, out_args):
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28 """
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29 Get file name prefix and create output directory
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31 Arguments:
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32 aligner_output : aligner output file or directory.
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33 out_args : dictionary of output arguments.
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35 Returns:
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36 str : file name prefix.
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37 """
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38 # Determine output directory
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39 if not out_args['out_dir']:
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40 out_dir = os.path.dirname(os.path.abspath(aligner_output))
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41 else:
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42 out_dir = os.path.abspath(out_args['out_dir'])
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43 if not os.path.exists(out_dir):
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44 os.mkdir(out_dir)
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45
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46 # Determine file prefix
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47 if out_args['out_name']:
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48 file_prefix = out_args['out_name']
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49 else:
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50 file_prefix = os.path.splitext(os.path.split(os.path.abspath(aligner_output))[1])[0]
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52 return os.path.join(out_dir, file_prefix)
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54
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55 def getSeqDict(seq_file):
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56 """
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57 Create a dictionary from a sequence file.
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59 Arguments:
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60 seq_file : sequence file.
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62 Returns:
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63 dict : sequence description as keys with Bio.SeqRecords as values.
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64 """
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65 seq_dict = SeqIO.to_dict(readSeqFile(seq_file),
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66 key_function=lambda x: x.description)
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67
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68 return seq_dict
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70
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71 def writeDb(db, fields, file_prefix, total_count, id_dict=None, no_parse=True, partial=False,
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72 out_args=default_out_args):
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73 """
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74 Writes tab-delimited database file in output directory.
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76 Arguments:
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77 db : a iterator of IgRecord objects containing alignment data.
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78 fields : a list of ordered field names to write.
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79 file_prefix : directory and prefix for CLIP tab-delim file.
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80 total_count : number of records (for progress bar).
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81 id_dict : a dictionary of the truncated sequence ID mapped to the full sequence ID.
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82 no_parse : if ID is to be parsed for pRESTO output with default delimiters.
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83 partial : if True put incomplete alignments in the pass file.
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84 out_args : common output argument dictionary from parseCommonArgs.
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85
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86 Returns:
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87 None
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88 """
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89 # Function to check for valid records strictly
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90 def _pass_strict(rec):
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91 valid = [rec.v_call and rec.v_call != 'None',
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92 rec.j_call and rec.j_call != 'None',
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93 rec.functional is not None,
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94 rec.seq_vdj,
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95 rec.junction]
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96 return all(valid)
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97
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98 # Function to check for valid records loosely
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99 def _pass_gentle(rec):
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100 valid = [rec.v_call and rec.v_call != 'None',
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101 rec.d_call and rec.d_call != 'None',
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102 rec.j_call and rec.j_call != 'None']
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103 return any(valid)
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104
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105 # Set pass criteria
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106 _pass = _pass_gentle if partial else _pass_strict
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107
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108 # Define output file names
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109 pass_file = '%s_db-pass.tab' % file_prefix
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110 fail_file = '%s_db-fail.tab' % file_prefix
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111
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112 # Initiate handles, writers and counters
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113 pass_handle = None
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114 fail_handle = None
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115 pass_writer = None
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116 fail_writer = None
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117 start_time = time()
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118 rec_count = pass_count = fail_count = 0
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119
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120 # Validate and write output
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121 printProgress(0, total_count, 0.05, start_time)
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122 for i, record in enumerate(db, start=1):
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123
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124 # Replace sequence description with full string, if required
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125 if id_dict is not None and record.id in id_dict:
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126 record.id = id_dict[record.id]
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127
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128 # Parse sequence description into new columns
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129 if not no_parse:
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130 try:
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131 record.annotations = parseAnnotation(record.id, delimiter=out_args['delimiter'])
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132 record.id = record.annotations['ID']
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133 del record.annotations['ID']
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134
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135 # TODO: This is not the best approach. should pass in output fields.
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136 # If first record, use parsed description to define extra columns
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137 if i == 1: fields.extend(list(record.annotations.keys()))
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138 except IndexError:
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139 # Could not parse pRESTO-style annotations so fall back to no parse
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140 no_parse = True
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141 sys.stderr.write('\nWARNING: Sequence annotation format not recognized. Sequence headers will not be parsed.\n')
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142
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143 # Count pass or fail and write to appropriate file
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144 if _pass(record):
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145 # Open pass file
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146 if pass_writer is None:
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147 pass_handle = open(pass_file, 'wt')
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148 pass_writer = getDbWriter(pass_handle, add_fields=fields)
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149
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150 # Write row to pass file
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151 pass_count += 1
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152 pass_writer.writerow(record.toDict())
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153 else:
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154 # Open failed file
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155 if out_args['failed'] and fail_writer is None:
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156 fail_handle = open(fail_file, 'wt')
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157 fail_writer = getDbWriter(fail_handle, add_fields=fields)
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158
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159 # Write row to fail file if specified
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160 fail_count += 1
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161 if fail_writer is not None:
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162 fail_writer.writerow(record.toDict())
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163
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164 # Print progress
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165 printProgress(i, total_count, 0.05, start_time)
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166
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167 # Print consol log
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168 log = OrderedDict()
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169 log['OUTPUT'] = pass_file
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170 log['PASS'] = pass_count
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171 log['FAIL'] = fail_count
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172 log['END'] = 'MakeDb'
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173 printLog(log)
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174
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175 if pass_handle is not None: pass_handle.close()
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176 if fail_handle is not None: fail_handle.close()
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177
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178
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179 # TODO: may be able to merge with other mains
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180 def parseIMGT(aligner_output, seq_file=None, no_parse=True, partial=False,
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181 parse_scores=False, parse_regions=False, parse_junction=False,
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182 out_args=default_out_args):
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183 """
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184 Main for IMGT aligned sample sequences.
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185
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186 Arguments:
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187 aligner_output : zipped file or unzipped folder output by IMGT.
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188 seq_file : FASTA file input to IMGT (from which to get seqID).
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189 no_parse : if ID is to be parsed for pRESTO output with default delimiters.
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190 partial : If True put incomplete alignments in the pass file.
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191 parse_scores : if True add alignment score fields to output file.
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192 parse_regions : if True add FWR and CDR region fields to output file.
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193 out_args : common output argument dictionary from parseCommonArgs.
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194
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195 Returns:
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196 None
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197 """
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198 # Print parameter info
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199 log = OrderedDict()
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200 log['START'] = 'MakeDb'
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201 log['ALIGNER'] = 'IMGT'
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202 log['ALIGNER_OUTPUT'] = aligner_output
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203 log['SEQ_FILE'] = os.path.basename(seq_file) if seq_file else ''
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204 log['NO_PARSE'] = no_parse
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205 log['PARTIAL'] = partial
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206 log['SCORES'] = parse_scores
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207 log['REGIONS'] = parse_regions
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208 log['JUNCTION'] = parse_junction
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209 printLog(log)
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210
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211 start_time = time()
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212 printMessage('Loading sequence files', start_time=start_time, width=25)
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213 # Extract IMGT files
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214 temp_dir, imgt_files = extractIMGT(aligner_output)
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215 # Count records in IMGT files
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216 total_count = countDbFile(imgt_files['summary'])
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217 # Get (parsed) IDs from fasta file submitted to IMGT
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218 id_dict = getIDforIMGT(seq_file) if seq_file else {}
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219 printMessage('Done', start_time=start_time, end=True, width=25)
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220
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221 # Parse IMGT output and write db
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222 with open(imgt_files['summary'], 'r') as summary_handle, \
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223 open(imgt_files['gapped'], 'r') as gapped_handle, \
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224 open(imgt_files['ntseq'], 'r') as ntseq_handle, \
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225 open(imgt_files['junction'], 'r') as junction_handle:
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226 parse_iter = IMGTReader(summary_handle, gapped_handle, ntseq_handle, junction_handle,
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227 parse_scores=parse_scores, parse_regions=parse_regions,
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228 parse_junction=parse_junction)
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229 file_prefix = getFilePrefix(aligner_output, out_args)
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230 writeDb(parse_iter, parse_iter.fields, file_prefix, total_count, id_dict=id_dict,
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231 no_parse=no_parse, partial=partial, out_args=out_args)
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232
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233 # Cleanup temp directory
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234 temp_dir.cleanup()
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235
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236 return None
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237
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238
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239 # TODO: may be able to merge with other mains
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240 def parseIgBLAST(aligner_output, seq_file, repo, no_parse=True, partial=False,
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241 parse_regions=False, parse_scores=False, parse_igblast_cdr3=False,
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242 out_args=default_out_args):
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243 """
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244 Main for IgBLAST aligned sample sequences.
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245
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246 Arguments:
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247 aligner_output : IgBLAST output file to process.
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248 seq_file : fasta file input to IgBlast (from which to get sequence).
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249 repo : folder with germline repertoire files.
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250 no_parse : if ID is to be parsed for pRESTO output with default delimiters.
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251 partial : If True put incomplete alignments in the pass file.
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252 parse_regions : if True add FWR and CDR fields to output file.
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253 parse_scores : if True add alignment score fields to output file.
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254 parse_igblast_cdr3 : if True parse CDR3 sequences generated by IgBLAST
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255 out_args : common output argument dictionary from parseCommonArgs.
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256
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257 Returns:
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258 None
0
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259 """
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260 # Print parameter info
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261 log = OrderedDict()
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262 log['START'] = 'MakeDB'
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263 log['ALIGNER'] = 'IgBlast'
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264 log['ALIGNER_OUTPUT'] = os.path.basename(aligner_output)
0
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265 log['SEQ_FILE'] = os.path.basename(seq_file)
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266 log['NO_PARSE'] = no_parse
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267 log['PARTIAL'] = partial
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268 log['SCORES'] = parse_scores
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269 log['REGIONS'] = parse_regions
0
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270 printLog(log)
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271
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272 start_time = time()
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273 printMessage('Loading sequence files', start_time=start_time, width=25)
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274 # Count records in sequence file
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275 total_count = countSeqFile(seq_file)
0
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276 # Get input sequence dictionary
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277 seq_dict = getSeqDict(seq_file)
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278 # Create germline repo dictionary
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279 repo_dict = readRepo(repo)
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280 printMessage('Done', start_time=start_time, end=True, width=25)
0
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281
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282 # Parse and write output
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283 with open(aligner_output, 'r') as f:
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284 parse_iter = IgBLASTReader(f, seq_dict, repo_dict,
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285 parse_scores=parse_scores, parse_regions=parse_regions,
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286 parse_igblast_cdr3=parse_igblast_cdr3)
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287 file_prefix = getFilePrefix(aligner_output, out_args)
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288 writeDb(parse_iter, parse_iter.fields, file_prefix, total_count,
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289 no_parse=no_parse, partial=partial, out_args=out_args)
0
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290
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291 return None
0
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292
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293
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294 # TODO: may be able to merge with other mains
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295 def parseIHMM(aligner_output, seq_file, repo, no_parse=True, partial=False,
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296 parse_scores=False, parse_regions=False, out_args=default_out_args):
0
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297 """
52
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298 Main for iHMMuneAlign aligned sample sequences.
0
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299
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300 Arguments:
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301 aligner_output : iHMMune-Align output file to process.
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302 seq_file : fasta file input to iHMMuneAlign (from which to get sequence).
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303 repo : folder with germline repertoire files.
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304 no_parse : if ID is to be parsed for pRESTO output with default delimiters.
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305 partial : If True put incomplete alignments in the pass file.
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306 parse_scores : if True parse alignment scores.
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307 parse_regions : if True add FWR and CDR region fields.
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308 out_args : common output argument dictionary from parseCommonArgs.
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309
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310 Returns:
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311 None
0
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312 """
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313 # Print parameter info
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314 log = OrderedDict()
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315 log['START'] = 'MakeDB'
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316 log['ALIGNER'] = 'iHMMune-Align'
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317 log['ALIGNER_OUTPUT'] = os.path.basename(aligner_output)
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318 log['SEQ_FILE'] = os.path.basename(seq_file)
0
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319 log['NO_PARSE'] = no_parse
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320 log['PARTIAL'] = partial
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321 log['SCORES'] = parse_scores
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322 log['REGIONS'] = parse_regions
0
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323 printLog(log)
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324
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325 start_time = time()
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326 printMessage('Loading sequence files', start_time=start_time, width=25)
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327 # Count records in sequence file
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328 total_count = countSeqFile(seq_file)
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329 # Get input sequence dictionary
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330 seq_dict = getSeqDict(seq_file)
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331 # Create germline repo dictionary
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332 repo_dict = readRepo(repo)
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333 printMessage('Done', start_time=start_time, end=True, width=25)
0
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334
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335 # Parse and write output
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336 with open(aligner_output, 'r') as f:
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337 parse_iter = IHMMuneReader(f, seq_dict, repo_dict,
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338 parse_scores=parse_scores, parse_regions=parse_regions)
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339 file_prefix = getFilePrefix(aligner_output, out_args)
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340 writeDb(parse_iter, parse_iter.fields, file_prefix, total_count,
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341 no_parse=no_parse, partial=partial, out_args=out_args)
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342
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343 return None
0
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344
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345
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346 def getArgParser():
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347 """
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348 Defines the ArgumentParser.
0
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349
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350 Returns:
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351 argparse.ArgumentParser
0
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352 """
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353 fields = dedent(
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354 '''
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355 output files:
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356 db-pass
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357 database of alignment records with functionality information,
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358 V and J calls, and a junction region.
0
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359 db-fail
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360 database with records that fail due to no functionality information
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361 (did not pass IMGT), no V call, no J call, or no junction region.
0
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362
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363 universal output fields:
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364 SEQUENCE_ID, SEQUENCE_INPUT, SEQUENCE_VDJ, SEQUENCE_IMGT,
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365 FUNCTIONAL, IN_FRAME, STOP, MUTATED_INVARIANT, INDELS,
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366 V_CALL, D_CALL, J_CALL,
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367 V_SEQ_START, V_SEQ_LENGTH,
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368 D_SEQ_START, D_SEQ_LENGTH, D_GERM_START, D_GERM_LENGTH,
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369 J_SEQ_START, J_SEQ_LENGTH, J_GERM_START, J_GERM_LENGTH,
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370 JUNCTION_LENGTH, JUNCTION, NP1_LENGTH, NP2_LENGTH,
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371 FWR1_IMGT, FWR2_IMGT, FWR3_IMGT, FWR4_IMGT,
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372 CDR1_IMGT, CDR2_IMGT, CDR3_IMGT
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373
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374 imgt specific output fields:
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375 V_GERM_START_IMGT, V_GERM_LENGTH_IMGT,
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376 N1_LENGTH, N2_LENGTH, P3V_LENGTH, P5D_LENGTH, P3D_LENGTH, P5J_LENGTH,
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377 D_FRAME, V_SCORE, V_IDENTITY, J_SCORE, J_IDENTITY,
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378
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379 igblast specific output fields:
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380 V_GERM_START_VDJ, V_GERM_LENGTH_VDJ,
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381 V_EVALUE, V_SCORE, V_IDENTITY, V_BTOP,
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382 J_EVALUE, J_SCORE, J_IDENTITY, J_BTOP.
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383 CDR3_IGBLAST_NT, CDR3_IGBLAST_AA
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384
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385 ihmm specific output fields:
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386 V_GERM_START_VDJ, V_GERM_LENGTH_VDJ,
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387 HMM_SCORE
0
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388 ''')
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389
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390 # Define ArgumentParser
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391 parser = ArgumentParser(description=__doc__, epilog=fields,
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392 formatter_class=CommonHelpFormatter)
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393 parser.add_argument('--version', action='version',
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394 version='%(prog)s:' + ' %s-%s' %(__version__, __date__))
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395 subparsers = parser.add_subparsers(title='subcommands', dest='command',
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396 help='Aligner used', metavar='')
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397 # TODO: This is a temporary fix for Python issue 9253
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398 subparsers.required = True
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399
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400 # Parent parser
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401 parser_parent = getCommonArgParser(seq_in=False, seq_out=False, log=False)
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402
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403 # IgBlast Aligner
52
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404 parser_igblast = subparsers.add_parser('igblast', parents=[parser_parent],
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405 formatter_class=CommonHelpFormatter,
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406 help='Process IgBLAST output.',
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parents: 0
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407 description='Process IgBLAST output.')
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408 parser_igblast.add_argument('-i', nargs='+', action='store', dest='aligner_outputs',
0
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409 required=True,
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410 help='''IgBLAST output files in format 7 with query sequence
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411 (IgBLAST argument \'-outfmt "7 std qseq sseq btop"\').''')
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412 parser_igblast.add_argument('-r', nargs='+', action='store', dest='repo', required=True,
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413 help='''List of folders and/or fasta files containing
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414 IMGT-gapped germline sequences corresponding to the
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415 set of germlines used in the IgBLAST alignment.''')
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416 parser_igblast.add_argument('-s', action='store', nargs='+', dest='seq_files',
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417 required=True,
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418 help='''List of input FASTA files (with .fasta, .fna or .fa
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parents: 0
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419 extension), containing sequences.''')
0
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420 parser_igblast.add_argument('--noparse', action='store_true', dest='no_parse',
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421 help='''Specify to prevent input sequence headers from being parsed
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422 to add new columns to database. Parsing of sequence headers requires
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423 headers to be in the pRESTO annotation format, so this should be specified
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424 when sequence headers are incompatible with the pRESTO annotation scheme.
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425 Note, unrecognized header formats will default to this behavior.''')
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426 parser_igblast.add_argument('--partial', action='store_true', dest='partial',
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427 help='''If specified, include incomplete V(D)J alignments in
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428 the pass file instead of the fail file.''')
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429 parser_igblast.add_argument('--scores', action='store_true', dest='parse_scores',
0
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430 help='''Specify if alignment score metrics should be
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431 included in the output. Adds the V_SCORE, V_IDENTITY,
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432 V_EVALUE, V_BTOP, J_SCORE, J_IDENTITY,
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433 J_BTOP, and J_EVALUE columns.''')
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434 parser_igblast.add_argument('--regions', action='store_true', dest='parse_regions',
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435 help='''Specify if IMGT FWR and CDRs should be
0
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436 included in the output. Adds the FWR1_IMGT, FWR2_IMGT,
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437 FWR3_IMGT, FWR4_IMGT, CDR1_IMGT, CDR2_IMGT, and
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438 CDR3_IMGT columns.''')
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439 parser_igblast.add_argument('--cdr3', action='store_true',
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440 dest='parse_igblast_cdr3',
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441 help='''Specify if the CDR3 sequences generated by IgBLAST
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442 should be included in the output. Adds the columns
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443 CDR3_IGBLAST_NT and CDR3_IGBLAST_AA. Requires IgBLAST
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444 version 1.5 or greater.''')
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445 parser_igblast.set_defaults(func=parseIgBLAST)
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446
0
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447 # IMGT aligner
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448 parser_imgt = subparsers.add_parser('imgt', parents=[parser_parent],
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449 formatter_class=CommonHelpFormatter,
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450 help='''Process IMGT/HighV-Quest output
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451 (does not work with V-QUEST).''',
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452 description='''Process IMGT/HighV-Quest output
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453 (does not work with V-QUEST).''')
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454 parser_imgt.add_argument('-i', nargs='+', action='store', dest='aligner_outputs',
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455 help='''Either zipped IMGT output files (.zip or .txz) or a
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456 folder containing unzipped IMGT output files (which must
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457 include 1_Summary, 2_IMGT-gapped, 3_Nt-sequences,
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458 and 6_Junction).''')
0
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459 parser_imgt.add_argument('-s', nargs='*', action='store', dest='seq_files',
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460 required=False,
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461 help='''List of input FASTA files (with .fasta, .fna or .fa
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462 extension) containing sequences.''')
0
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463 parser_imgt.add_argument('--noparse', action='store_true', dest='no_parse',
52
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464 help='''Specify to prevent input sequence headers from being parsed
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465 to add new columns to database. Parsing of sequence headers requires
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466 headers to be in the pRESTO annotation format, so this should be specified
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467 when sequence headers are incompatible with the pRESTO annotation scheme.
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468 Note, unrecognized header formats will default to this behavior.''')
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469 parser_imgt.add_argument('--partial', action='store_true', dest='partial',
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470 help='''If specified, include incomplete V(D)J alignments in
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471 the pass file instead of the fail file.''')
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472 parser_imgt.add_argument('--scores', action='store_true', dest='parse_scores',
0
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473 help='''Specify if alignment score metrics should be
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474 included in the output. Adds the V_SCORE, V_IDENTITY,
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475 J_SCORE and J_IDENTITY.''')
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476 parser_imgt.add_argument('--regions', action='store_true', dest='parse_regions',
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477 help='''Specify if IMGT FWRs and CDRs should be
0
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478 included in the output. Adds the FWR1_IMGT, FWR2_IMGT,
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479 FWR3_IMGT, FWR4_IMGT, CDR1_IMGT, CDR2_IMGT, and
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480 CDR3_IMGT columns.''')
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481 parser_imgt.add_argument('--junction', action='store_true', dest='parse_junction',
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482 help='''Specify if detailed junction fields should be
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483 included in the output. Adds the columns
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484 N1_LENGTH, N2_LENGTH, P3V_LENGTH, P5D_LENGTH, P3D_LENGTH,
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485 P5J_LENGTH, D_FRAME.''')
0
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486 parser_imgt.set_defaults(func=parseIMGT)
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487
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488 # iHMMuneAlign Aligner
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489 parser_ihmm = subparsers.add_parser('ihmm', parents=[parser_parent],
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490 formatter_class=CommonHelpFormatter,
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491 help='Process iHMMune-Align output.',
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492 description='Process iHMMune-Align output.')
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493 parser_ihmm.add_argument('-i', nargs='+', action='store', dest='aligner_outputs',
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494 required=True,
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495 help='''iHMMune-Align output file.''')
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496 parser_ihmm.add_argument('-r', nargs='+', action='store', dest='repo', required=True,
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497 help='''List of folders and/or FASTA files containing
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498 IMGT-gapped germline sequences corresponding to the
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davidvanzessen
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499 set of germlines used in the IgBLAST alignment.''')
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500 parser_ihmm.add_argument('-s', action='store', nargs='+', dest='seq_files',
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501 required=True,
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502 help='''List of input FASTA files (with .fasta, .fna or .fa
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503 extension) containing sequences.''')
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504 parser_ihmm.add_argument('--noparse', action='store_true', dest='no_parse',
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505 help='''Specify to prevent input sequence headers from being parsed
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parents: 0
diff changeset
506 to add new columns to database. Parsing of sequence headers requires
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507 headers to be in the pRESTO annotation format, so this should be specified
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davidvanzessen
parents: 0
diff changeset
508 when sequence headers are incompatible with the pRESTO annotation scheme.
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509 Note, unrecognized header formats will default to this behavior.''')
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510 parser_ihmm.add_argument('--partial', action='store_true', dest='partial',
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511 help='''If specified, include incomplete V(D)J alignments in
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512 the pass file instead of the fail file.''')
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513 parser_ihmm.add_argument('--scores', action='store_true', dest='parse_scores',
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514 help='''Specify if alignment score metrics should be
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515 included in the output. Adds the path score of the
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516 iHMMune-Align hidden Markov model to HMM_SCORE.''')
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517 parser_ihmm.add_argument('--regions', action='store_true', dest='parse_regions',
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518 help='''Specify if IMGT FWRs and CDRs should be
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davidvanzessen
parents: 0
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519 included in the output. Adds the FWR1_IMGT, FWR2_IMGT,
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520 FWR3_IMGT, FWR4_IMGT, CDR1_IMGT, CDR2_IMGT, and
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521 CDR3_IMGT columns.''')
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522 parser_ihmm.set_defaults(func=parseIHMM)
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523
0
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524 return parser
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525
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526
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527 if __name__ == "__main__":
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528 """
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529 Parses command line arguments and calls main
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530 """
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531 parser = getArgParser()
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532 args = parser.parse_args()
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533 args_dict = parseCommonArgs(args, in_arg='aligner_outputs')
0
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534
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535 # Set no ID parsing if sequence files are not provided
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536 if 'seq_files' in args_dict and not args_dict['seq_files']:
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537 args_dict['no_parse'] = True
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538
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539 # Delete
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540 if 'seq_files' in args_dict: del args_dict['seq_files']
52
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541 if 'aligner_outputs' in args_dict: del args_dict['aligner_outputs']
0
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542 if 'command' in args_dict: del args_dict['command']
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543 if 'func' in args_dict: del args_dict['func']
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544
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545 if args.command == 'imgt':
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546 for i in range(len(args.__dict__['aligner_outputs'])):
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547 args_dict['aligner_output'] = args.__dict__['aligner_outputs'][i]
0
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548 args_dict['seq_file'] = args.__dict__['seq_files'][i] \
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549 if args.__dict__['seq_files'] else None
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550 args.func(**args_dict)
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551 elif args.command == 'igblast' or args.command == 'ihmm':
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552 for i in range(len(args.__dict__['aligner_outputs'])):
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553 args_dict['aligner_output'] = args.__dict__['aligner_outputs'][i]
0
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diff changeset
554 args_dict['seq_file'] = args.__dict__['seq_files'][i]
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555 args.func(**args_dict)