diff shm_csr.py @ 47:64711f461c8e draft

Uploaded
author davidvanzessen
date Thu, 04 May 2017 07:43:09 -0400
parents 77a7ac76c7b9
children c5295dd10dfc
line wrap: on
line diff
--- a/shm_csr.py	Wed Apr 12 04:28:16 2017 -0400
+++ b/shm_csr.py	Thu May 04 07:43:09 2017 -0400
@@ -1,287 +1,436 @@
-from __future__ import division
+import argparse
+import logging
+import sys
+import os
+import re
+
 from collections import defaultdict
-import re
-import argparse
+
+def main():
+	parser = argparse.ArgumentParser()
+	parser.add_argument("--input", help="The '7_V-REGION-mutation-and-AA-change-table' and '10_V-REGION-mutation-hotspots' merged together, with an added 'best_match' annotation")
+	parser.add_argument("--genes", help="The genes available in the 'best_match' column")
+	parser.add_argument("--empty_region_filter", help="Where does the sequence start?", choices=['leader', 'FR1', 'CDR1', 'FR2'])
+	parser.add_argument("--output", help="Output file")
 
-parser = argparse.ArgumentParser()
-parser.add_argument("--input",
-					help="The '7_V-REGION-mutation-and-AA-change-table' and '10_V-REGION-mutation-hotspots' merged together, with an added 'best_match' annotation")
-parser.add_argument("--genes", help="The genes available in the 'best_match' column")
-parser.add_argument("--empty_region_filter", help="Where does the sequence start?", choices=['leader', 'FR1', 'CDR1', 'FR2'])
-parser.add_argument("--output", help="Output file")
+	args = parser.parse_args()
+
+	infile = args.input
+	genes = str(args.genes).split(",")
+	empty_region_filter = args.empty_region_filter
+	outfile = args.output
 
-args = parser.parse_args()
+	genedic = dict()
 
-infile = args.input
-genes = str(args.genes).split(",")
-empty_region_filter = args.empty_region_filter
-outfile = args.output
-
-genedic = dict()
+	mutationdic = dict()
+	mutationMatcher = re.compile("^(.)(\d+).(.),?(.)?(\d+)?.?(.)?(.?.?.?.?.?)?")
+	NAMatchResult = (None, None, None, None, None, None, '')
+	geneMatchers = {gene: re.compile("^" + gene + ".*") for gene in genes}
+	linecount = 0
 
-mutationdic = dict()
-mutationMatcher = re.compile("^(.)(\d+).(.),?(.)?(\d+)?.?(.)?(.?.?.?.?.?)?")
-NAMatchResult = (None, None, None, None, None, None, '')
-linecount = 0
+	IDIndex = 0
+	best_matchIndex = 0
+	fr1Index = 0
+	cdr1Index = 0
+	fr2Index = 0
+	cdr2Index = 0
+	fr3Index = 0
+	first = True
+	IDlist = []
+	mutationList = []
+	mutationListByID = {}
+	cdr1LengthDic = {}
+	cdr2LengthDic = {}
+
+	fr1LengthDict = {}
+	fr2LengthDict = {}
+	fr3LengthDict = {}
+
+	cdr1LengthIndex = 0
+	cdr2LengthIndex = 0
 
-IDIndex = 0
-best_matchIndex = 0
-fr1Index = 0
-cdr1Index = 0
-fr2Index = 0
-cdr2Index = 0
-fr3Index = 0
-first = True
-IDlist = []
-mutationList = []
-mutationListByID = {}
-cdr1LengthDic = {}
-cdr2LengthDic = {}
+	fr1SeqIndex = 0
+	fr2SeqIndex = 0
+	fr3SeqIndex = 0
+
+	tandem_sum_by_class = defaultdict(int)
+	expected_tandem_sum_by_class = defaultdict(float)
 
-with open(infile, 'ru') as i:
-	for line in i:
-		if first:
+	with open(infile, 'ru') as i:
+		for line in i:
+			if first:
+				linesplt = line.split("\t")
+				IDIndex = linesplt.index("Sequence.ID")
+				best_matchIndex = linesplt.index("best_match")
+				fr1Index = linesplt.index("FR1.IMGT")
+				cdr1Index = linesplt.index("CDR1.IMGT")
+				fr2Index = linesplt.index("FR2.IMGT")
+				cdr2Index = linesplt.index("CDR2.IMGT")
+				fr3Index = linesplt.index("FR3.IMGT")
+				cdr1LengthIndex = linesplt.index("CDR1.IMGT.seq")
+				cdr2LengthIndex = linesplt.index("CDR2.IMGT.seq")
+				fr1SeqIndex = linesplt.index("FR1.IMGT.seq")
+				fr2SeqIndex = linesplt.index("FR2.IMGT.seq")
+				fr3SeqIndex = linesplt.index("FR3.IMGT.seq")
+				first = False
+				continue
+			linecount += 1
 			linesplt = line.split("\t")
-			IDIndex = linesplt.index("Sequence.ID")
-			best_matchIndex = linesplt.index("best_match")
-			fr1Index = linesplt.index("FR1.IMGT")
-			cdr1Index = linesplt.index("CDR1.IMGT")
-			fr2Index = linesplt.index("FR2.IMGT")
-			cdr2Index = linesplt.index("CDR2.IMGT")
-			fr3Index = linesplt.index("FR3.IMGT")
-			cdr1LengthIndex = linesplt.index("CDR1.IMGT.length")
-			cdr2LengthIndex = linesplt.index("CDR2.IMGT.length")
-			first = False
-			continue
-		linecount += 1
-		linesplt = line.split("\t")
-		ID = linesplt[IDIndex]
-		genedic[ID] = linesplt[best_matchIndex]
-		try:
-			mutationdic[ID + "_FR1"] = [mutationMatcher.match(x).groups() for x in linesplt[fr1Index].split("|") if x] if (linesplt[fr1Index] != "NA" and empty_region_filter == "leader") else []
-			mutationdic[ID + "_CDR1"] = [mutationMatcher.match(x).groups() for x in linesplt[cdr1Index].split("|") if x] if (linesplt[cdr1Index] != "NA" and empty_region_filter in ["leader", "FR1"]) else []
-			mutationdic[ID + "_FR2"] = [mutationMatcher.match(x).groups() for x in linesplt[fr2Index].split("|") if x] if (linesplt[fr2Index] != "NA" and empty_region_filter in ["leader", "FR1", "CDR1"]) else []
-			mutationdic[ID + "_CDR2"] = [mutationMatcher.match(x).groups() for x in linesplt[cdr2Index].split("|") if x] if (linesplt[cdr2Index] != "NA") else []
-			mutationdic[ID + "_FR2-CDR2"] = mutationdic[ID + "_FR2"] + mutationdic[ID + "_CDR2"]
-			mutationdic[ID + "_FR3"] = [mutationMatcher.match(x).groups() for x in linesplt[fr3Index].split("|") if x] if linesplt[fr3Index] != "NA" else []
-		except Exception as e:
-			print "Something went wrong while processing this line:"
-			print linesplt
-			print linecount
-			print e
-		mutationList += mutationdic[ID + "_FR1"] + mutationdic[ID + "_CDR1"] + mutationdic[ID + "_FR2"] + mutationdic[ID + "_CDR2"] + mutationdic[ID + "_FR3"]
-		mutationListByID[ID] = mutationdic[ID + "_FR1"] + mutationdic[ID + "_CDR1"] + mutationdic[ID + "_FR2"] + mutationdic[ID + "_CDR2"] + mutationdic[ID + "_FR3"]
+			ID = linesplt[IDIndex]
+			genedic[ID] = linesplt[best_matchIndex]
+			try:
+				mutationdic[ID + "_FR1"] = [mutationMatcher.match(x).groups() for x in linesplt[fr1Index].split("|") if x] if (linesplt[fr1Index] != "NA" and empty_region_filter == "leader") else []
+				mutationdic[ID + "_CDR1"] = [mutationMatcher.match(x).groups() for x in linesplt[cdr1Index].split("|") if x] if (linesplt[cdr1Index] != "NA" and empty_region_filter in ["leader", "FR1"]) else []
+				mutationdic[ID + "_FR2"] = [mutationMatcher.match(x).groups() for x in linesplt[fr2Index].split("|") if x] if (linesplt[fr2Index] != "NA" and empty_region_filter in ["leader", "FR1", "CDR1"]) else []
+				mutationdic[ID + "_CDR2"] = [mutationMatcher.match(x).groups() for x in linesplt[cdr2Index].split("|") if x] if (linesplt[cdr2Index] != "NA") else []
+				mutationdic[ID + "_FR2-CDR2"] = mutationdic[ID + "_FR2"] + mutationdic[ID + "_CDR2"]
+				mutationdic[ID + "_FR3"] = [mutationMatcher.match(x).groups() for x in linesplt[fr3Index].split("|") if x] if linesplt[fr3Index] != "NA" else []
+			except Exception as e:
+				print "Something went wrong while processing this line:"
+				print linesplt
+				print linecount
+				print e
+			mutationList += mutationdic[ID + "_FR1"] + mutationdic[ID + "_CDR1"] + mutationdic[ID + "_FR2"] + mutationdic[ID + "_CDR2"] + mutationdic[ID + "_FR3"]
+			mutationListByID[ID] = mutationdic[ID + "_FR1"] + mutationdic[ID + "_CDR1"] + mutationdic[ID + "_FR2"] + mutationdic[ID + "_CDR2"] + mutationdic[ID + "_FR3"]
+
+			cdr1Length = len(linesplt[cdr1LengthIndex])
+			cdr2Length = len(linesplt[cdr2LengthIndex])
+
+			#print linesplt[fr2SeqIndex]
+			fr1Length = len(linesplt[fr1SeqIndex]) if empty_region_filter == "leader" else 0
+			fr2Length = len(linesplt[fr2SeqIndex]) if empty_region_filter in ["leader", "FR1", "CDR1"] else 0
+			fr3Length = len(linesplt[fr3SeqIndex])
+
+			cdr1LengthDic[ID] = cdr1Length
+			cdr2LengthDic[ID] = cdr2Length
+
+			fr1LengthDict[ID] = fr1Length
+			fr2LengthDict[ID] = fr2Length
+			fr3LengthDict[ID] = fr3Length
+
+			IDlist += [ID]
+
+
+	#tandem mutation stuff
+	tandem_frequency = defaultdict(int)
+	mutation_frequency = defaultdict(int)
+
+	tandem_file = os.path.join(os.path.dirname(outfile), "tandems_by_id.txt")
+	with open(tandem_file, 'w') as o:
+		highest_tandem_length = 0
+
+		o.write("Sequence.ID\tnumber_of_mutations\tnumber_of_tandems\tregion_length\texpected_tandems\tlongest_tandem\ttandems\n")
+		for ID in IDlist:
+			mutations = mutationListByID[ID]
+			if len(mutations) == 0:
+				continue
+			last_mut = max(mutations, key=lambda x: int(x[1]))
+
+			last_mut_pos = int(last_mut[1])
+
+			mut_positions = [False] * (last_mut_pos + 1)
+
+			for mutation in mutations:
+				frm, where, to, frmAA, whereAA, toAA, thing = mutation
+				where = int(where)
+				mut_positions[where] = True
+
+			tandem_muts = []
+			tandem_start = -1
+			tandem_length = 0
+			for i in range(len(mut_positions)):
+				if mut_positions[i]:
+					if tandem_start == -1:
+						tandem_start = i
+					tandem_length += 1
+					#print "".join(["1" if x else "0" for x in mut_positions[:i+1]])
+				else:
+					if tandem_length > 1:
+						tandem_muts.append((tandem_start, tandem_length))
+						#print "{0}{1} {2}:{3}".format(" " * (i - tandem_length), "^" * tandem_length, tandem_start, tandem_length)
+					tandem_start = -1
+					tandem_length = 0
+			if tandem_length > 1:  # if the sequence ends with a tandem mutation
+				tandem_muts.append((tandem_start, tandem_length))
+
+			if len(tandem_muts) > 0:
+				if highest_tandem_length < len(tandem_muts):
+					highest_tandem_length = len(tandem_muts)
 
-		cdr1Length = linesplt[cdr1LengthIndex]
-		cdr2Length = linesplt[cdr2LengthIndex]
+			region_length = fr1LengthDict[ID] + cdr1LengthDic[ID] + fr2LengthDict[ID] + cdr2LengthDic[ID] + fr3LengthDict[ID]
+			longest_tandem = max(tandem_muts, key=lambda x: x[1]) if len(tandem_muts) else (0, 0)
+			num_mutations = len(mutations)
+			f_num_mutations = float(num_mutations)
+			num_tandem_muts = len(tandem_muts)
+			expected_tandem_muts = f_num_mutations * (f_num_mutations - 1.0) / float(region_length)
+			o.write("{0}\t{1}\t{2}\t{3}\t{4}\t{5}\t{6}\n".format(ID,
+																str(num_mutations),
+																str(num_tandem_muts),
+																str(region_length),
+																str(round(expected_tandem_muts, 2)),
+																str(longest_tandem[1]),
+																str(tandem_muts)))
+			gene = genedic[ID]
+			if gene.find("unmatched") == -1:
+				tandem_sum_by_class[gene] += num_tandem_muts
+				expected_tandem_sum_by_class[gene] += expected_tandem_muts
 
-		cdr1LengthDic[ID] = int(cdr1Length) if cdr1Length != "X" else 0
-		cdr2LengthDic[ID] = int(cdr2Length) if cdr2Length != "X" else 0
-		
-		IDlist += [ID]
+				tandem_sum_by_class["all"] += num_tandem_muts
+				expected_tandem_sum_by_class["all"] += expected_tandem_muts
 
-#print mutationList, linecount
+				gene = gene[:3]
+				if gene in ["IGA", "IGG"]:
+					tandem_sum_by_class[gene] += num_tandem_muts
+					expected_tandem_sum_by_class[gene] += expected_tandem_muts
+			else:
+				tandem_sum_by_class["unmatched"] += num_tandem_muts
+				expected_tandem_sum_by_class["unmatched"] += expected_tandem_muts
+
 
-AALength = (int(max(mutationList, key=lambda i: int(i[4]) if i[4] else 0)[4]) + 1)  # [4] is the position of the AA mutation, None if silent
-if AALength < 60:
-	AALength = 64
+			for tandem_mut in tandem_muts:
+				tandem_frequency[str(tandem_mut[1])] += 1
+			#print "\t".join([ID, str(len(tandem_muts)), str(longest_tandem[1]) , str(tandem_muts)])
+
+	tandem_freq_file = os.path.join(os.path.dirname(outfile), "tandem_frequency.txt")
+	with open(tandem_freq_file, 'w') as o:
+		for frq in sorted([int(x) for x in tandem_frequency.keys()]):
+			o.write("{0}\t{1}\n".format(frq, tandem_frequency[str(frq)]))
 
-AA_mutation = [0] * AALength
-AA_mutation_dic = {"IGA": AA_mutation[:], "IGG": AA_mutation[:], "IGM": AA_mutation[:], "IGE": AA_mutation[:], "unm": AA_mutation[:], "all": AA_mutation[:]}
-AA_mutation_empty = AA_mutation[:]
+	tandem_row = []
+	print genes
+	print tandem_sum_by_class
+	print expected_tandem_sum_by_class
+	genes_extra = list(genes)
+	genes_extra.append("all")
+	for x, y, in zip([tandem_sum_by_class[x] for x in genes_extra], [expected_tandem_sum_by_class[x] for x in genes_extra]):
+		if y != 0:
+			tandem_row += [x, round(y, 2), round(x / y, 2)]
+		else:
+			tandem_row += [x, round(y, 2), 0]
+	
+	"""
+	print tandem_row
+	tandem_row += tandem_row[-3:]
+	print tandem_row
+	all_expected_tandem = expected_tandem_sum_by_class["all"]
+	all_tandem = tandem_sum_by_class["all"]
+	if all_expected_tandem == 0:
+		tandem_row[-6:-3] = [all_tandem, round(all_expected_tandem, 2), 0]
+	else:
+		tandem_row[-6:-3] = [all_tandem, round(all_expected_tandem, 2), round(all_tandem / all_expected_tandem, 2)]
+	print tandem_row
+	"""
+	for i in range(len(genes_extra)):
+		gene = genes_extra[i]
+		print gene, tandem_row[i*3:i*3+3]
 
-aa_mutations_by_id_file = outfile[:outfile.rindex("/")] + "/aa_id_mutations.txt"
-with open(aa_mutations_by_id_file, 'w') as o:
-	o.write("ID\tbest_match\t" + "\t".join([str(x) for x in range(1,AALength)]) + "\n")
-	for ID in mutationListByID.keys():
-		AA_mutation_for_ID = AA_mutation_empty[:]
-		for mutation in mutationListByID[ID]:
-			if mutation[4]:
-				AA_mutation_position = int(mutation[4])
-				AA_mutation[AA_mutation_position] += 1
-				AA_mutation_for_ID[AA_mutation_position] += 1
-				clss = genedic[ID][:3]
-				AA_mutation_dic[clss][AA_mutation_position] += 1
-		o.write(ID + "\t" + genedic[ID] + "\t" + "\t".join([str(x) for x in AA_mutation_for_ID[1:]]) + "\n")
+	tandem_freq_file = os.path.join(os.path.dirname(outfile), "shm_overview_tandem_row.txt")
+	with open(tandem_freq_file, 'w') as o:
+		o.write("Tandems/Expected (ratio),{0}\n".format(",".join([str(x) for x in tandem_row])))
+
+	#print mutationList, linecount
+
+	AALength = (int(max(mutationList, key=lambda i: int(i[4]) if i[4] else 0)[4]) + 1)  # [4] is the position of the AA mutation, None if silent
+	if AALength < 60:
+		AALength = 64
+
+	AA_mutation = [0] * AALength
+	AA_mutation_dic = {"IGA": AA_mutation[:], "IGG": AA_mutation[:], "IGM": AA_mutation[:], "IGE": AA_mutation[:], "unm": AA_mutation[:], "all": AA_mutation[:]}
+	AA_mutation_empty = AA_mutation[:]
+
+	aa_mutations_by_id_file = outfile[:outfile.rindex("/")] + "/aa_id_mutations.txt"
+	with open(aa_mutations_by_id_file, 'w') as o:
+		o.write("ID\tbest_match\t" + "\t".join([str(x) for x in range(1,AALength)]) + "\n")
+		for ID in mutationListByID.keys():
+			AA_mutation_for_ID = AA_mutation_empty[:]
+			for mutation in mutationListByID[ID]:
+				if mutation[4]:
+					AA_mutation_position = int(mutation[4])
+					AA_mutation[AA_mutation_position] += 1
+					AA_mutation_for_ID[AA_mutation_position] += 1
+					clss = genedic[ID][:3]
+					AA_mutation_dic[clss][AA_mutation_position] += 1
+			o.write(ID + "\t" + genedic[ID] + "\t" + "\t".join([str(x) for x in AA_mutation_for_ID[1:]]) + "\n")
 
 
 
-#absent AA stuff
-absentAACDR1Dic = defaultdict(list)
-absentAACDR1Dic[5] = range(29,36)
-absentAACDR1Dic[6] = range(29,35)
-absentAACDR1Dic[7] = range(30,35)
-absentAACDR1Dic[8] = range(30,34)
-absentAACDR1Dic[9] = range(31,34)
-absentAACDR1Dic[10] = range(31,33)
-absentAACDR1Dic[11] = [32]
+	#absent AA stuff
+	absentAACDR1Dic = defaultdict(list)
+	absentAACDR1Dic[5] = range(29,36)
+	absentAACDR1Dic[6] = range(29,35)
+	absentAACDR1Dic[7] = range(30,35)
+	absentAACDR1Dic[8] = range(30,34)
+	absentAACDR1Dic[9] = range(31,34)
+	absentAACDR1Dic[10] = range(31,33)
+	absentAACDR1Dic[11] = [32]
 
-absentAACDR2Dic = defaultdict(list)
-absentAACDR2Dic[0] = range(55,65)
-absentAACDR2Dic[1] = range(56,65)
-absentAACDR2Dic[2] = range(56,64)
-absentAACDR2Dic[3] = range(57,64)
-absentAACDR2Dic[4] = range(57,63)
-absentAACDR2Dic[5] = range(58,63)
-absentAACDR2Dic[6] = range(58,62)
-absentAACDR2Dic[7] = range(59,62)
-absentAACDR2Dic[8] = range(59,61)
-absentAACDR2Dic[9] = [60]
+	absentAACDR2Dic = defaultdict(list)
+	absentAACDR2Dic[0] = range(55,65)
+	absentAACDR2Dic[1] = range(56,65)
+	absentAACDR2Dic[2] = range(56,64)
+	absentAACDR2Dic[3] = range(57,64)
+	absentAACDR2Dic[4] = range(57,63)
+	absentAACDR2Dic[5] = range(58,63)
+	absentAACDR2Dic[6] = range(58,62)
+	absentAACDR2Dic[7] = range(59,62)
+	absentAACDR2Dic[8] = range(59,61)
+	absentAACDR2Dic[9] = [60]
 
-absentAA = [len(IDlist)] * (AALength-1)
-for k, cdr1Length in cdr1LengthDic.iteritems():
-	for c in absentAACDR1Dic[cdr1Length]:
-		absentAA[c] -= 1
+	absentAA = [len(IDlist)] * (AALength-1)
+	for k, cdr1Length in cdr1LengthDic.iteritems():
+		for c in absentAACDR1Dic[cdr1Length]:
+			absentAA[c] -= 1
 
-for k, cdr2Length in cdr2LengthDic.iteritems():
-	for c in absentAACDR2Dic[cdr2Length]:
-		absentAA[c] -= 1
+	for k, cdr2Length in cdr2LengthDic.iteritems():
+		for c in absentAACDR2Dic[cdr2Length]:
+			absentAA[c] -= 1
 
 
-aa_mutations_by_id_file = outfile[:outfile.rindex("/")] + "/absent_aa_id.txt"
-with open(aa_mutations_by_id_file, 'w') as o:
-	o.write("ID\tcdr1length\tcdr2length\tbest_match\t" + "\t".join([str(x) for x in range(1,AALength)]) + "\n")
-	for ID in IDlist:
-		absentAAbyID = [1] * (AALength-1)
-		cdr1Length = cdr1LengthDic[ID]
-		for c in absentAACDR1Dic[cdr1Length]:
-			absentAAbyID[c] -= 1
+	aa_mutations_by_id_file = outfile[:outfile.rindex("/")] + "/absent_aa_id.txt"
+	with open(aa_mutations_by_id_file, 'w') as o:
+		o.write("ID\tcdr1length\tcdr2length\tbest_match\t" + "\t".join([str(x) for x in range(1,AALength)]) + "\n")
+		for ID in IDlist:
+			absentAAbyID = [1] * (AALength-1)
+			cdr1Length = cdr1LengthDic[ID]
+			for c in absentAACDR1Dic[cdr1Length]:
+				absentAAbyID[c] -= 1
 
-		cdr2Length = cdr2LengthDic[ID]
-		for c in absentAACDR2Dic[cdr2Length]:
-			absentAAbyID[c] -= 1
-		o.write(ID + "\t" + str(cdr1Length) + "\t" + str(cdr2Length) + "\t" + genedic[ID] + "\t" + "\t".join([str(x) for x in absentAAbyID]) + "\n")
+			cdr2Length = cdr2LengthDic[ID]
+			for c in absentAACDR2Dic[cdr2Length]:
+				absentAAbyID[c] -= 1
+			o.write(ID + "\t" + str(cdr1Length) + "\t" + str(cdr2Length) + "\t" + genedic[ID] + "\t" + "\t".join([str(x) for x in absentAAbyID]) + "\n")
 
-if linecount == 0:
-	print "No data, exiting"
-	with open(outfile, 'w') as o:
-		o.write("RGYW (%)," + ("0,0,0\n" * len(genes)))
-		o.write("WRCY (%)," + ("0,0,0\n" * len(genes)))
-		o.write("WA (%)," + ("0,0,0\n" * len(genes)))
-		o.write("TW (%)," + ("0,0,0\n" * len(genes)))
-	import sys
+	if linecount == 0:
+		print "No data, exiting"
+		with open(outfile, 'w') as o:
+			o.write("RGYW (%)," + ("0,0,0\n" * len(genes)))
+			o.write("WRCY (%)," + ("0,0,0\n" * len(genes)))
+			o.write("WA (%)," + ("0,0,0\n" * len(genes)))
+			o.write("TW (%)," + ("0,0,0\n" * len(genes)))
+		import sys
 
-	sys.exit()
+		sys.exit()
 
-hotspotMatcher = re.compile("[actg]+,(\d+)-(\d+)\((.*)\)")
-RGYWCount = {}
-WRCYCount = {}
-WACount = {}
-TWCount = {}
+	hotspotMatcher = re.compile("[actg]+,(\d+)-(\d+)\((.*)\)")
+	RGYWCount = {}
+	WRCYCount = {}
+	WACount = {}
+	TWCount = {}
 
-#IDIndex = 0
-ataIndex = 0
-tatIndex = 0
-aggctatIndex = 0
-atagcctIndex = 0
-first = True
-with open(infile, 'ru') as i:
-	for line in i:
-		if first:
+	#IDIndex = 0
+	ataIndex = 0
+	tatIndex = 0
+	aggctatIndex = 0
+	atagcctIndex = 0
+	first = True
+	with open(infile, 'ru') as i:
+		for line in i:
+			if first:
+				linesplt = line.split("\t")
+				ataIndex = linesplt.index("X.a.t.a")
+				tatIndex = linesplt.index("t.a.t.")
+				aggctatIndex = linesplt.index("X.a.g.g.c.t..a.t.")
+				atagcctIndex = linesplt.index("X.a.t..a.g.c.c.t.")
+				first = False
+				continue
 			linesplt = line.split("\t")
-			ataIndex = linesplt.index("X.a.t.a")
-			tatIndex = linesplt.index("t.a.t.")
-			aggctatIndex = linesplt.index("X.a.g.g.c.t..a.t.")
-			atagcctIndex = linesplt.index("X.a.t..a.g.c.c.t.")
-			first = False
-			continue
-		linesplt = line.split("\t")
-		gene = linesplt[best_matchIndex]
-		ID = linesplt[IDIndex]
-		RGYW = [(int(x), int(y), z) for (x, y, z) in
-				[hotspotMatcher.match(x).groups() for x in linesplt[aggctatIndex].split("|") if x]]
-		WRCY = [(int(x), int(y), z) for (x, y, z) in
-				[hotspotMatcher.match(x).groups() for x in linesplt[atagcctIndex].split("|") if x]]
-		WA = [(int(x), int(y), z) for (x, y, z) in
-			  [hotspotMatcher.match(x).groups() for x in linesplt[ataIndex].split("|") if x]]
-		TW = [(int(x), int(y), z) for (x, y, z) in
-			  [hotspotMatcher.match(x).groups() for x in linesplt[tatIndex].split("|") if x]]
-		RGYWCount[ID], WRCYCount[ID], WACount[ID], TWCount[ID] = 0, 0, 0, 0
+			gene = linesplt[best_matchIndex]
+			ID = linesplt[IDIndex]
+			RGYW = [(int(x), int(y), z) for (x, y, z) in
+					[hotspotMatcher.match(x).groups() for x in linesplt[aggctatIndex].split("|") if x]]
+			WRCY = [(int(x), int(y), z) for (x, y, z) in
+					[hotspotMatcher.match(x).groups() for x in linesplt[atagcctIndex].split("|") if x]]
+			WA = [(int(x), int(y), z) for (x, y, z) in
+				[hotspotMatcher.match(x).groups() for x in linesplt[ataIndex].split("|") if x]]
+			TW = [(int(x), int(y), z) for (x, y, z) in
+				[hotspotMatcher.match(x).groups() for x in linesplt[tatIndex].split("|") if x]]
+			RGYWCount[ID], WRCYCount[ID], WACount[ID], TWCount[ID] = 0, 0, 0, 0
 
-		mutationList = mutationdic[ID + "_FR1"] + mutationdic[ID + "_CDR1"] + mutationdic[ID + "_FR2"] + mutationdic[ID + "_CDR2"] + mutationdic[ID + "_FR3"]
-		for mutation in mutationList:
-			frm, where, to, AAfrm, AAwhere, AAto, junk = mutation
-			mutation_in_RGYW = any([(start <= int(where) <= end) for (start, end, region) in RGYW])
-			mutation_in_WRCY = any([(start <= int(where) <= end) for (start, end, region) in WRCY])
-			mutation_in_WA = any([(start <= int(where) <= end) for (start, end, region) in WA])
-			mutation_in_TW = any([(start <= int(where) <= end) for (start, end, region) in TW])
+			mutationList = mutationdic[ID + "_FR1"] + mutationdic[ID + "_CDR1"] + mutationdic[ID + "_FR2"] + mutationdic[ID + "_CDR2"] + mutationdic[ID + "_FR3"]
+			for mutation in mutationList:
+				frm, where, to, AAfrm, AAwhere, AAto, junk = mutation
+				mutation_in_RGYW = any([(start <= int(where) <= end) for (start, end, region) in RGYW])
+				mutation_in_WRCY = any([(start <= int(where) <= end) for (start, end, region) in WRCY])
+				mutation_in_WA = any([(start <= int(where) <= end) for (start, end, region) in WA])
+				mutation_in_TW = any([(start <= int(where) <= end) for (start, end, region) in TW])
 
-			in_how_many_motifs = sum([mutation_in_RGYW, mutation_in_WRCY, mutation_in_WA, mutation_in_TW])
+				in_how_many_motifs = sum([mutation_in_RGYW, mutation_in_WRCY, mutation_in_WA, mutation_in_TW])
 
-			if in_how_many_motifs > 0:
-				RGYWCount[ID] += (1.0 * int(mutation_in_RGYW)) / in_how_many_motifs
-				WRCYCount[ID] += (1.0 * int(mutation_in_WRCY)) / in_how_many_motifs
-				WACount[ID] += (1.0 * int(mutation_in_WA)) / in_how_many_motifs
-				TWCount[ID] += (1.0 * int(mutation_in_TW)) / in_how_many_motifs
+				if in_how_many_motifs > 0:
+					RGYWCount[ID] += (1.0 * int(mutation_in_RGYW)) / in_how_many_motifs
+					WRCYCount[ID] += (1.0 * int(mutation_in_WRCY)) / in_how_many_motifs
+					WACount[ID] += (1.0 * int(mutation_in_WA)) / in_how_many_motifs
+					TWCount[ID] += (1.0 * int(mutation_in_TW)) / in_how_many_motifs
 
 
-def mean(lst):
-	return (float(sum(lst)) / len(lst)) if len(lst) > 0 else 0.0
+	def mean(lst):
+		return (float(sum(lst)) / len(lst)) if len(lst) > 0 else 0.0
 
 
-def median(lst):
-	lst = sorted(lst)
-	l = len(lst)
-	if l == 0:
-		return 0
-	if l == 1:
-		return lst[0]
+	def median(lst):
+		lst = sorted(lst)
+		l = len(lst)
+		if l == 0:
+			return 0
+		if l == 1:
+			return lst[0]
+			
+		l = int(l / 2)
 		
-	l = int(l / 2)
-	
-	if len(lst) % 2 == 0:
-		return float(lst[l] + lst[(l - 1)]) / 2.0
-	else:
-		return lst[l]
-
-funcs = {"mean": mean, "median": median, "sum": sum}
+		if len(lst) % 2 == 0:
+			return float(lst[l] + lst[(l - 1)]) / 2.0
+		else:
+			return lst[l]
 
-directory = outfile[:outfile.rfind("/") + 1]
-value = 0
-valuedic = dict()
+	funcs = {"mean": mean, "median": median, "sum": sum}
 
-for fname in funcs.keys():
-	for gene in genes:
-		with open(directory + gene + "_" + fname + "_value.txt", 'r') as v:
-			valuedic[gene + "_" + fname] = float(v.readlines()[0].rstrip())
-	with open(directory + "all_" + fname + "_value.txt", 'r') as v:
-		valuedic["total_" + fname] = float(v.readlines()[0].rstrip())
-	
+	directory = outfile[:outfile.rfind("/") + 1]
+	value = 0
+	valuedic = dict()
 
-def get_xyz(lst, gene, f, fname):
-	x = round(round(f(lst), 1))
-	y = valuedic[gene + "_" + fname]
-	z = str(round(x / float(y) * 100, 1)) if y != 0 else "0"
-	return (str(x), str(y), z)
+	for fname in funcs.keys():
+		for gene in genes:
+			with open(directory + gene + "_" + fname + "_value.txt", 'r') as v:
+				valuedic[gene + "_" + fname] = float(v.readlines()[0].rstrip())
+		with open(directory + "all_" + fname + "_value.txt", 'r') as v:
+			valuedic["total_" + fname] = float(v.readlines()[0].rstrip())
+		
 
-dic = {"RGYW": RGYWCount, "WRCY": WRCYCount, "WA": WACount, "TW": TWCount}
-arr = ["RGYW", "WRCY", "WA", "TW"]
+	def get_xyz(lst, gene, f, fname):
+		x = round(round(f(lst), 1))
+		y = valuedic[gene + "_" + fname]
+		z = str(round(x / float(y) * 100, 1)) if y != 0 else "0"
+		return (str(x), str(y), z)
 
-geneMatchers = {gene: re.compile("^" + gene + ".*") for gene in genes}
+	dic = {"RGYW": RGYWCount, "WRCY": WRCYCount, "WA": WACount, "TW": TWCount}
+	arr = ["RGYW", "WRCY", "WA", "TW"]
 
-for fname in funcs.keys():
-	func = funcs[fname]
-	foutfile = outfile[:outfile.rindex("/")] + "/hotspot_analysis_" + fname + ".txt"
-	with open(foutfile, 'w') as o:
-		for typ in arr:
-			o.write(typ + " (%)")
-			curr = dic[typ]
-			for gene in genes:
-				geneMatcher = geneMatchers[gene]
-				if valuedic[gene + "_" + fname] is 0:
-					o.write(",0,0,0")
-				else:
-					x, y, z = get_xyz([curr[x] for x in [y for y, z in genedic.iteritems() if geneMatcher.match(z)]], gene, func, fname)
-					o.write("," + x + "," + y + "," + z)
-			x, y, z = get_xyz([y for x, y in curr.iteritems() if not genedic[x].startswith("unmatched")], "total", func, fname)
-			#x, y, z = get_xyz([y for x, y in curr.iteritems()], "total", func, fname)
-			o.write("," + x + "," + y + "," + z + "\n")
+	for fname in funcs.keys():
+		func = funcs[fname]
+		foutfile = outfile[:outfile.rindex("/")] + "/hotspot_analysis_" + fname + ".txt"
+		with open(foutfile, 'w') as o:
+			for typ in arr:
+				o.write(typ + " (%)")
+				curr = dic[typ]
+				for gene in genes:
+					geneMatcher = geneMatchers[gene]
+					if valuedic[gene + "_" + fname] is 0:
+						o.write(",0,0,0")
+					else:
+						x, y, z = get_xyz([curr[x] for x in [y for y, z in genedic.iteritems() if geneMatcher.match(z)]], gene, func, fname)
+						o.write("," + x + "," + y + "," + z)
+				x, y, z = get_xyz([y for x, y in curr.iteritems() if not genedic[x].startswith("unmatched")], "total", func, fname)
+				#x, y, z = get_xyz([y for x, y in curr.iteritems()], "total", func, fname)
+				o.write("," + x + "," + y + "," + z + "\n")
 
 
-# for testing
-seq_motif_file = outfile[:outfile.rindex("/")] + "/motif_per_seq.txt"
-with open(seq_motif_file, 'w') as o:
-	o.write("ID\tRGYW\tWRCY\tWA\tTW\n")
-	for ID in IDlist:
-		#o.write(ID + "\t" + str(round(RGYWCount[ID], 2)) + "\t" + str(round(WRCYCount[ID], 2)) + "\t" + str(round(WACount[ID], 2)) + "\t" + str(round(TWCount[ID], 2)) + "\n")
-		o.write(ID + "\t" + str(RGYWCount[ID]) + "\t" + str(WRCYCount[ID]) + "\t" + str(WACount[ID]) + "\t" + str(TWCount[ID]) + "\n")
+	# for testing
+	seq_motif_file = outfile[:outfile.rindex("/")] + "/motif_per_seq.txt"
+	with open(seq_motif_file, 'w') as o:
+		o.write("ID\tRGYW\tWRCY\tWA\tTW\n")
+		for ID in IDlist:
+			#o.write(ID + "\t" + str(round(RGYWCount[ID], 2)) + "\t" + str(round(WRCYCount[ID], 2)) + "\t" + str(round(WACount[ID], 2)) + "\t" + str(round(TWCount[ID], 2)) + "\n")
+			o.write(ID + "\t" + str(RGYWCount[ID]) + "\t" + str(WRCYCount[ID]) + "\t" + str(WACount[ID]) + "\t" + str(TWCount[ID]) + "\n")
+
+if __name__ == "__main__":
+	main()