diff aggregate.py @ 18:13f7c0c33dcb draft

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author kaymccoy
date Fri, 12 Aug 2016 23:01:17 -0400
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--- /dev/null	Thu Jan 01 00:00:00 1970 +0000
+++ b/aggregate.py	Fri Aug 12 23:01:17 2016 -0400
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+# A translation of aggregate.pl into python! For analysis of Tn-Seq.
+# This script requires BioPython just like calc_fitness.py, so you need it installed along with its dependencies if you want to run these scripts on your own.
+# How to install BioPython and a list of its dependencies can be found here: http://biopython.org/DIST/docs/install/Installation.html
+# K. McCoy
+
+
+
+
+
+
+
+
+
+##### ARGUMENTS #####
+
+def print_usage():
+	print "Aggregate.py's usage is as follows:" + "\n\n"
+	print "\033[1m" + "Required" + "\033[0m" + "\n"
+	print "-o" + "\t\t" + "Output file for aggregated data." + "\n"
+	print "\n"
+	print "\033[1m" + "Optional" + "\033[0m" + "\n"
+	print "-c" + "\t\t" + "Check for missing genes in the data set - provide a reference genome in genbank format. Missing genes will be sent to stdout." + "\n"
+	print "-m" + "\t\t" + "Place a mark in an extra column for this set of genes. Provide a file with a list of genes seperated by newlines." + "\n"
+	print "-x" + "\t\t" + "Cutoff: Don't include fitness scores with average counts (c1+c2)/2 < x (default: 0)" + "\n"
+	print "-b" + "\t\t" + "Blanks: Exclude -b % of blank fitness scores (scores where c2 = 0) (default: 0 = 0%)" + "\n"
+	print "-f" + "\t\t" + "An in-between file carrying information on the blank count found from calc_fitness or consol_fitness; one of two ways to pass a blank count to this script" + "\n"
+	print "-w" + "\t\t" + "Use weighted algorithm to calculate averages, variance, sd, se" + "\n"
+	print "-l" + "\t\t" + "Weight ceiling: maximum value to use as a weight (default: 999,999)" + "\n"
+	print "\n"
+	print "All remainder arguements will be treated as fitness files (those files created by calc_fitness.py)" + "\n"
+	print "\n"
+	
+import argparse 
+parser = argparse.ArgumentParser()
+parser.add_argument("-o", action="store", dest="summary")
+parser.add_argument("-c", action="store", dest="find_missing")
+parser.add_argument("-m", action="store", dest="marked")
+parser.add_argument("-x", action="store", dest="cutoff")
+parser.add_argument("-b", action="store", dest="blank_pc")
+parser.add_argument("-f", action="store", dest="blank_file")
+parser.add_argument("-w", action="store", dest="weighted")
+parser.add_argument("-l", action="store", dest="weight_ceiling")
+parser.add_argument("fitnessfiles", nargs=argparse.REMAINDER)
+
+arguments = parser.parse_args()
+
+if not arguments.summary:
+	print "\n" + "You are missing a value for the -o flag. "
+	print_usage() 
+	quit()
+
+if not arguments.fitnessfiles:
+	print "\n" + "You are missing fitness file(s); these should be entered immediately after all the flags. "
+	print_usage() 
+	quit()
+	
+# 999,999 is a trivial placeholder number
+	
+if (not arguments.weight_ceiling):
+	arguments.weight_ceiling = 999999
+	
+# Cutoff exists to discard positions with a low number of counted transcripts, because their fitness may not be as accurate - for the same reasoning that studies with low sample sizes can be innacurate. 
+	
+if (not arguments.cutoff):
+	arguments.cutoff = 0
+
+# Gets information from the txt output file of calc_fit / consol, if inputted
+
+if arguments.blank_file:
+	with open(arguments.blank_file) as file:
+		blank_pc = file.read().splitlines()
+		arguments.blank_pc = float(blank_pc[0].split()[1])
+
+if (not arguments.blank_pc):
+	arguments.blank_pc = 0
+
+
+
+
+
+##### SUBROUTINES #####
+
+# A subroutine that calculates the average, variance, standard deviation (sd), and standard error (se) of a group of scores; for use when aggregating scores by gene later on
+
+import math
+def average(scores):
+	sum = 0
+	num = 0
+	for i in scores:
+		sum += float(i)
+		num += 1
+	average = sum/num
+	xminusxbars = 0
+	for i in scores:
+		xminusxbars += (i - average)**2
+	variance = xminusxbars/(num-1)
+	sd = math.sqrt(variance)
+	se = sd / math.sqrt(num)
+	return (average, variance, sd, se)
+
+# A subroutine that calculates the weighted average, variance, standard deviation (sd), and standard error (se) of a group of scores; the weights come from the number of reads each insertion location has
+# For use when aggregating scores by gene later on, if the weighted argument is called
+
+def weighted_average(scores,weights):
+	sum = 0
+	weighted_average = 0
+	weighted_variance = 0
+	top = 0
+	bottom = 0
+	i = 0
+	while i < len(weights):
+		if not scores[i]:
+			scores[i] = 0.0
+		top += float(weights[i])*float(scores[i])
+		bottom += float(weights[i])
+		i += 1
+	if bottom == 0:
+		return 0
+	weighted_average = top/bottom
+	top = 0
+	bottom = 0
+	i = 0
+	while i < len(weights):
+		top += float(weights[i]) * (float(scores[i]) - weighted_average)**2
+		bottom += float(weights[i])
+		i += 1
+	weighted_variance = top/bottom
+ 	weighted_stdev = math.sqrt(weighted_variance)
+	weighted_stder = weighted_stdev/math.sqrt(len(scores))
+	return (weighted_average, weighted_variance, weighted_stdev, weighted_stder)
+
+
+
+
+
+
+
+
+
+
+##### AGGREGATION / CALCULATIONS #####
+
+#Reads the genes which should be marked in the final aggregate file into an array
+
+import os.path
+if arguments.marked:
+	with open(arguments.marked) as file:
+		marked_set = file.read().splitlines()
+
+#Creates a dictionary of dictionaries to contain a summary of all genes and their fitness values
+#The fitness values and weights match up, so that the weight of gene_summary[locus]["w"][2] would be gene_summary[locus]["s"][2]
+
+import csv
+gene_summary = {}
+for eachfile in arguments.fitnessfiles:
+	with open(eachfile) as csvfile:
+		lines = csv.reader(csvfile)
+		for line in lines:
+			locus = line[9]
+			w = line[12]
+			if w == 'nW':
+				continue
+			if not w:
+				w == 0
+			c1 = float(line[2])
+			c2 = float(line[3])
+			avg = (c1+c2)/2
+			if avg < float(arguments.cutoff):
+				continue
+			if avg > float(arguments.weight_ceiling):
+				avg = arguments.weight_ceiling
+			if locus not in gene_summary:
+				gene_summary[locus] = {"w" : [], "s": []}
+			gene_summary[locus]["w"].append(w)
+			gene_summary[locus]["s"].append(avg)
+
+#If finding any missing gene loci is requested in the arguments, starts out by loading all the known features from a genbank file
+
+from Bio import SeqIO
+if (arguments.find_missing):
+	output = [["locus","mean","var","sd","se","gene","Total","Blank","Not Blank","Blank Removed","M\n"]]
+	handle = open(arguments.find_missing, "rU")
+	for record in SeqIO.parse(handle, "genbank"):
+		refname = record.id
+		features = record.features
+	handle.close()
+	
+#Goes through the features to find which are genes
+	
+	for feature in features:
+		gene = ""
+		if feature.type == "gene":
+			locus = "".join(feature.qualifiers["locus_tag"])
+			if "gene" in feature.qualifiers:
+				gene = "".join(feature.qualifiers["gene"])
+		else:
+			continue
+			
+#Goes through the fitness scores of insertions within each gene, and removes whatever % of blank fitness scores were requested along with their corresponding weights
+
+		sum = 0
+		num = 0
+		avgsum = 0
+		blank_ws = 0
+		i = 0
+		if locus in gene_summary.keys():
+			for w in gene_summary[locus]["w"]:
+				if float(w) == 0:
+					blank_ws += 1
+				else:
+					sum += float(w)
+					num += 1
+			count = num + blank_ws		
+			removed = 0
+			to_remove = int(float(arguments.blank_pc)*count)
+			if blank_ws > 0:
+				i = 0
+				while i < len(gene_summary[locus]["w"]):
+					w = gene_summary[locus]["w"][i] 
+					if removed == to_remove:
+						break
+					if float(w) == 0:
+						del gene_summary[locus]["w"][i]
+						del gene_summary[locus]["s"][i]
+						removed += 1
+						i -= 1
+					i += 1
+			
+#If all the fitness values within a gene are empty, sets mean/var to 0.10 and Xs out sd/se; marks the gene if that's requested
+
+			if num == 0:	
+				if (arguments.marked and locus in marked_set):
+					output.append([locus, "0.10", "0.10", "X", "X", gene, count, blank_ws, num, removed, "M", "\n"])
+				else:
+					output.append([locus, "0.10", "0.10", "X", "X", gene, count, blank_ws, num, removed, "\n"])
+
+#Otherwise calls average() or weighted_average() to find the aggregate w / count / standard deviation / standard error of the insertions within each gene; marks the gene if that's requested
+
+			else:
+				if not arguments.weighted:
+					(average, variance, stdev, stderr) = average(gene_summary[locus]["w"])
+				else:
+					(average, variance, stdev, stderr) = weighted_average(gene_summary[locus]["w"],gene_summary[locus]["s"])
+				if (arguments.marked and locus in marked_set):
+					output.append([locus, average, variance, stdev, stderr, gene, count, blank_ws, num, removed, "M", "\n"])
+				else:
+					output.append([locus, average, variance, stdev, stderr, gene, count, blank_ws, num, removed, "\n"])
+		
+#If a gene doesn't have any insertions, sets mean/var to 0.10 and Xs out sd/se, plus leaves count through removed blank because there were no reads.
+
+		else:
+			if (arguments.marked and locus in marked_set):
+				output.append([locus, "0.10", "0.10", "X", "X", gene, "", "", "", "", "M", "\n"])
+			else:
+				output.append([locus, "0.10", "0.10", "X", "X", gene, "", "", "", "", "\n"])
+
+#Writes the aggregated fitness file
+
+	with open(arguments.summary, "wb") as csvfile:
+		writer = csv.writer(csvfile)
+		writer.writerows(output)
+
+#If finding missing genes is not requested, just finds the aggregate w / count / standard deviation / standard error of the insertions within each gene, and writes them to a file, plus marks the genes requested
+#This is never called through Galaxy since finding missing genes is just better than not finding them.
+
+else:
+	output = [["Locus","W","Count","SD","SE","M\n"]]
+	for gene in gene_summary.keys():
+		sum = 0
+		num = 0
+		average = 0
+		if "w" not in gene_summary[gene]:
+			continue
+		for i in gene_summary[gene]["w"]:
+			sum += i
+			num += 1
+		average = sum/num
+		xminusxbars = 0
+		for i in w:
+			xminusxbars += (i-average)**2
+		if num > 1:
+			sd = math.sqrt(xminusxbars/(num-1))
+			se = sd / math.sqrt(num)
+		if (arguments.marked and locus in marked_set):
+			output.append([gene, average, num, sd, se, "M", "\n"])
+		else:
+			output.append([gene, average, num, sd, se, "\n"])
+	with open(arguments.summary, "wb") as csvfile:
+		writer = csv.writer(csvfile)
+		writer.writerows(output)
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+#
+#                                                ~MMM=:DMMM?,              +NMMO=,:~I8MMMMM8+, ,                   ~I8MMMMMN87~?8NNMMN8:                                  +NMND~        +MN=  ,$MMMI        ?M8,          ,OM8,     :MN+    =MM?                                                                               ,MMDNMMD                            ,+DM8I,                     ,,:::~~~::::::::::
+#                                                IMMMNMM8I                  ,I8MM87~::+$8NNMMMMOI+=~~:,  ,,:~=?$DNMMMMMMDOZI7ZDMMMD8I                                       ,  ,        $M8+?8MM8I ,        7MI           +MN=       ZMN,   8MD                                                                                 MMN8MMM,                             :$ONM8I+=:,              ,,,::::~~~~~=====~:
+#                    ,                           ,DMNN7:      ,                ,OMMN7==~::~=?8NNMMMMMMNNMMMMMMMMMMMN8OO8ODNMMMMMD~               ,                                      IMMNMN~            ,OM+          ,NM$        ,NMO, :MM$                                            ,               ,:::,,::::,,         $MNMMNM,                ,,,             :?ONMMNN8?~,, ,       ,,,,,,,::~~=+++??=~
+#          ,,:=+????+,                                        I$                 :ZMMN8$~:,,,  ,:=?7$O8DD8O$7+==+$O8DNMMMMMMMMM$                  ?$,             ==                     ,~,               ~NM=          8MD,         ,OM8ZMMO                                             ,               ,::::::~~~:,,         ?MNMMZ                ,,,,,,               ,+7ONMNMD8O$+~,       ,,,,,,::~====::
+#    ,:=IONMMMMMMMMMMM8:                                      ZN$:                 ,~DMMMND7=,       ,,:~====:=$DNMMMMMMMN88MMMZ                 +N$   ,      ,7DN8=                                       =MN,         IMM             =DMN7                                                               ,,,,,,,,,,            ,~?,                  ,,,,,                 ,   ~?$8MMMMNN8Z?~:,,       ,:::,,,
+#+ONMMMMMMNO7=:,, ,,+MMO,                                      7D$:                     ~OMNMNNMNNNNNNNNNNNMMMMMMMMMNMMMM?,~MMM8                 ND, 7MM=,   ?NN:,                                         +MO,        ?MM,                                                                                    ,,,,                                                                   , :,:=$DMMMMMMN87=~,  ,,,,,
+#MMND8$: ,          7NM7                                      , =?~,,,                     :?88DDDNNMNNNDNDD88Z?:,   ZMM$  ,MMMO                ,MM+ZNMM? ::ZNZ,                                           +M$        ,MM?,                                                                                                                                                           ,   ,, ,,:=?Z8NMMMMMN8Z+=, 
+#:               ~ZMM8~                                  ,,,,                                                        8MM,  +MMM,                 7ZZI~=$OOZ$:            ,:+???+,                          +MZ       =MN=                                                                                      ,,,                                                   ,       ,,:~=?IIII$ZO88DDNNNNNNMMMMMMMMMMN~,
+#             ,:OMMM?                                   ,,:,,                                                        8MM  ~NNM7                                        :OMMMMMMMMO:                        =M8,     :NM~                                     =              ,~?I,                              ,,,,,,,,,,,,,,                                           ,~$DNMMMMMMMMMND8O888Z$II7777I??+++===:, 
+#            ?DMM8?,                                                                                                 OMN??NMM$                                       ~8MMMO?===7MMM8:                      ~NM=    =NN:                                    ,OM~   ,,       +NMMN~                             ,,,,,    ,,,,,,,,                                  ,?$O8NNMDDZ7?+=~:,  ,              ,           ,
+#       , ~$MMMD+                    ,                  ,                                                            ~MMNMMN~                                     : +NMMZ,       NMNM~                      7MI , ZNN,,                                    IMN, :DMNM+  ~+NMMMD~,                          ,,,,,,,,  ,, ,,,,,:,                                +OMNMMOI~,                                        
+#     , $MMMM$,                 , ,=?ODNMNNMNMMMMNNND~                                                                ,$D$, ,                                     ,8MM8           ,MMM7                     ,ZMNNMM=                                       DMMNMMMMMMMMMMMMNI:  ,                         ,,,,,,,,,         ,,,                              ?NMO=, ,                                            
+#   ,~ZNMND7,          ,,:~=+$DNNMMMMNDDDD888OZZZZ8NMMN                                                                                                           IMMN:            ,MMN~                      +Z$+,                                        ?NNNDO+:?O888OI,                                ,,,,,,,                           ,,,            +MN+                                                 
+# =ONMMM8~    ,  ,:=IDMMMMMMMND8$+:, ,           ,INMNZ                                                                                                          :MMM~,          ,  +MMD                                                                         , ,,                                     ,,::,,,,                         ,,:::,           ?M8:                    ,                            
+#8MMMNZ~   , =I$ONMMNDZ7?+,                 ,,=I8NMD7:                                                                                                           DMMN                DMN=                                                                                                                 ,:::,                          ,:~~=~~:,,         :ZMMZI+:                ,,            ,,,,,,,        
+#MN7:,:=7ONMMMD$?=,                  ,  ~7ODNMM$+:                                                                                                              ~MMM++7ZOZOO8O8D8$~  ,MM8                                                                                                                                           ,,,,::~~==~~:,,            :+7DMMMMNNDD88OOZZZO88DNNNN8=,     ,,,,,,,        
+#I,~+DMMNND7:   ,                   ,$MMMMMN7,                                                                                                                  $MMN :??+=~::,,,,     NMD,                                                                                                                                           ,:,,:::~=~~,,,,                ,,=I$8DNMMMMMMMMMNMMMNZ:       ,,,,,,        
+#DNMNN7:,                     ,+ONMMMMNI:                                                                                                                       NMM$                  DMN=                                                                                                                                                 ,,,,:::::~~::,,                ?DMMMMDZ=,                ,,,          
+#N8$: ,,,               ,:=?ONMMMD8Z+,                              ,,,,                                                                                        MMM=                  ZMMI                                                                                                                                               ,=?$8NMMMMMMMMMMMN87=~~,,,  :=ZMMMMD$~                                  
+#     ,             ,=ZNMMMMMNI:,                               ,~?Z88888$=,         ,:~+??~,                                                                   MMM,                  IMM$                                                                                                                                          ,=ZNMMMMMN8$+~=~=~~===7ODNMMMN8DNMMMN+,                        ,,            
+#        ,     ~?$ODMMMMNZ?:                                  :II+~,     ,=7=    :?77?=:====?O+             ,,:,,,                                              MMM,                  ?MM$                                                           ,,,,                                                                       :?ONMM8II=,                ,  =DMMMM87=,                          ,,,,           
+#    , ,, ,~I8MMMMMMN87?~:=+?7$ZOO88DD888O$I+~:,            ~ZZ:           ,$7,~??,          ,?+         ,+Z8$?==??=                                            MMM                   =MM$                                                    :?ODNNNNNNNMMO:                                                               ,:?NNMNO=                     ,,,IMMMNZ,                 ,,,:,,,,,,,,,,,,,           
+#,   ,~7DNMMMMMMMMMMMNNNNMMMNND8Z7II7$$$$ZODNNNMND$,      :O$:           , ,IN$,               I+     +ZZ=,,       7+                                           MMM   =ODDDDNNNNNN8=  :MM$                                               ?DDNN8?::,,  ,,7NMM8,                                                             7NMNZ~,                        :OMMM$, ,                 ,,:::~~:,,,,,,,,,,,,,,,,,    
+#?8NMMMMMMMMMMMN8I:,,,                      ~$NMMM$     :87               +DM$                 ID+~78I,            O7   :=+~                                    MMM    ,   ,          ?MM7                                           $NMN$,      :I8MMMMMMNMDNNNNNNNNNDD88ZI=:                                          ,ZMM7,                           +MMN~                        ,,,:::,,     ,,:,::::,,,,,,
+#MMMD8DNMDZ7=:                      ,:=+7ZNNNDOZ~      =DI ,            :7MM7,                 IDDZI,             =DN88ZI77$N?                                  NMM:                  $MM=                             ~:         =OMNZ+    ,=7DMMMMMMNDDOOOOZ$7IIIII77$ZOO8NNMMD$+~                                   :OND~, ,,                         MMN=                                      ,,,,,,,,,,,,, 
+#:     ,,,               ,    ,:+ZNMNNMMNO?:          +8?              +NMN7                    ,                  ,        ON~                                 8MM$                  NMD,                          ,7MMMN,     7MMO,   ,:ONMMMMN8I:                           ,~ZNMNN$,                               ~MM? ,                            ZMNND?~:             ,,, ,,                 ,,,,,,      
+# ,         ,      ,:~?7$ZZ8NNMMMNO7I=,              ~D+               ZNO=,                        ,                       :NM$:                               =MMM                  NMO                          ~NMNMMM   :ZDN$, ,,=7DMD$?=,                                     :?ZMD$:                            :DMNOZZZ$:          ,,,,,          ~IDMMMMMMMMMMMMMMMMMMMMM8~                             
+#       ,        =DMNNNNNNNN8$=                       :                  ,                   ,,~?ODNZ:                        :DM?     =:                       ,MMM7                +MN~                         ,MM8:NMN  INMZ, ,=ONOI                 ,                              +NMZ                       ,,+ZDMMMMMMMN+,        ,,,,,,,            ,~?$ODNNNNNNNMMMMMMMMD=                            ,
+#                ,:::::~7$I?=:                                                          ,~78NMNMMMMM?           :+Z+            M8    $MM,  =,                   DMMD                NMN                          DMO OMM77NMI, ?8$~      , ~ZDDDNNNNNMMMMMMMMNMNNNNNDD8Z+:,              IMN:                I8DNMMMMN7~:                 ,,,,,,          +$O$,       ~$DMMMMMMMD~              ,,             ,
+# :I+   :ID?           ~ZDNNNNMMNO?~,,                                         ,:::::=7ONMNNNDMMMN$       ,=IONNMMMMZ:,        ~MO  ,7MMMI $MN, ,                :MMN   ?8NDDNND$~  +MM~                         ,MM~=MMMNMD,            =ONNDDNMMMMMMMMMMMMNNND88DNNNNNMMMMMMMNDO7+~::,:,7MM+             =DMMMNNO?                                    ~ZMMMMM7::~INMMMMMMMMMMMMMN8:                            
+#:MMM+,INMMM:               ,:~+78NMMMM8?==++++++???++??I$Z77$$$$$$$7II??I$ZZO8MMMMM8Z7~     ,IZI       ~77+   ?MMMMD88MN8+~,      +8MO$OMMMMMNMMMZ,     ~=:      OMM?             ,MM?                          ZMM~MMMM8~ ,:+7$$$$ZZ7?==:   :8MDOZ$ZZZODMMMMMM8+,          ,:=?$ZZOOOOOOZ$:               ,                                 ,     ,=8MNMMMMMMMMMMNDZ$7$MMMMMMMMMMMMMMMMMMNI+?I7ZDNMMMMMMMMMM$, 
+#=MMMZ$MMMMMM~    ,::::~==~~+I$8NMMMMMN$::::::::,,,   ,,=ONDDO$7II?+~,           ,                                     ,,$DD87:  =NND=   ,  ,+$$=:~,     ,:,      ,MMD             NMI    ,~?Z8DND88$?:          8MM$MMMZ:=~:,~~:::,,,=$DNMND$MM8,          ~DMNMMMN?,           :7$7?+==~=:,,,,,,,,     ,,,,,,, ,,,,,,::,,,,,:::::::,,,::::::,,   ,OM$,7MMMMMMNZ=  ,    ~8MMMMMMMNDO7I7MMMMMMMMMMMMN8Z7+?NMMMD,,
+#NMMMMMM? ,++,                  ~=I??=                                                                                                                        :7$  7MM+           7M7                      ,:?ZZ$MMM8NMZ~+,               :?INMMM?             +$NMMMNZ:          :+?I7$$Z$O88D88DDDDNNNNNNNNNNNNNNNNNNDDNDDDNNNNNMMNMMMMMMMNNNN$:??~   ,+II?=,            , ,,         ,?I??+=~,        :MMMMMNM
+#MMMMMD~                                                                      ,:=++++++++++=~,,,                                                            +MMN:  ,MMD,         :M$                 ,INMNMMMMMMMMMM~~~?D,                    :OMM8:     ,+$8Z$+,,$MMMMMD,                                                                                                                                :IODDDD
+#N$~,,,                                                          ,~I$8NMMMMMMMMMNMMMMMMMMMMMMMMMMNNNN7:                                                   ZMMMM?    NMMMNZ~:,   +$:                ,NMNI:::~?8MMM7I?   IO      ~I$ODNNNDND8OO$I?DMMMD$I8NMMMMMMMNMNMMMMMMM8=,                                                                                                                              ,     
+#                                                        ,:~?ONMMMMNNNDDD8NMD+,                ,~?Z8NMNM8=,                                           , $MNNNMM?   :DMMMNDD8DZ7$+,                 ,8NMMNMNMMMD$:      =8, ,INMMMNZI?====+I$$8DDNMMMMMMNNNMND7,    :~$DMMMNMO                                                                                                                                    
+#                                                    ,?DMMMMMNNNMMM+::~++ZMMMM8ZZZZZZ$II77II7???=~:,  ,+DMN7                                         =NMM8, ,NMMNMNMN$?I7I77OZ~                         ~8D$~,         ,MMMMMMNM8:                 :DMMM, ,             DMO=ZMZ,                                                                                                                                 
+#                                                 ,?OMMN87=,     IZNNNMMMMMMMMMMND7IIII??III$8DDNN8Z+, , ~MM                                       ?OMMMM~  ,MMMMMZ+                                   ~I=              IMMMO$$=                     ?NNM?               MN~ +NM:                                                                                                                                
+#                                                IMM8~             =MD,        ?N=         :,,      ,+77?=+,                                    $8MM7::OMMMMMMZ+                                     I+,                  ?NI   ,                      :MMM8$            NNI,  7M$                                                                                                                               
+#                                               ,MMN               $MD         7M+                     ~ODNZ~,                               :7MMN? ,  $MMMN=             7MNMN:                   +8+                     ,D8~                         =MMDD8Z=       =NMD     OM7,                                                                                                                             
+#                                                OMMD~             IMN:        =8O:,~+$OO?  :IZ+:          :ZMMNM8=                        =DMMI    ~8MM8=     , ,,     :8M?,$MM,     ::,         DM,       =Z~     ,I8DDDN8$DMN~                        MN    ,=Z8DNDZZ=        MD:                                                                                                  ,                          
+#                 ,,,,,,,,                        ?8NNMM8$I????++=+8MMMD$77$ZDMMNMMMDNOZ~     :IZ8DOI~,     =MMMNMNM8I:,                 ,7M8I    +MND7       ~MNDDM8~  7MI   MM7  :8MDDM7       8N=     ~ID$,   ,:Z$?:,,,=ONNMNMD=                     ?M$                ::    +MI                                                                    ,,,,,,::~~:::,,,,,,, ,+OMO:,  ,                          
+#       ,,,::~===~~:,                                ,~+I$88DNNNNNNNNNDNDD8O$?~:,                   ,7DNMNDMMMN~,ZMND$8NNDZ=,            $M?    ?NNN7        =MO    OMD$MM=   MMMINMO   $M7     DN:    8MMD:    ?D~        :O8ZDMMMMD=,               ,ZN$              ,INNMD=  :NO                                                                                     ,:+8MMMMMMMMDI:,::,,                    
+#=~~~::~~::,                                                                                            ~?78MMMDNMM?    :+I$DD87?=,      OMNDO$7$OI:         7N+     ?MNMN~   MMMNM$    7M$    7M$   +NMMO,    ?N~        ~8+   ~7NMMNNOI=         ~?ONZ+             ~ZNND,$M8  ~MO                                                                                       $MMMMMMMMMM7:::~~~~~~=+++===~~~:::,,  
+#                                                                                                            ,~ONMMNO~,        :?8NNMN8?~ZNMMMMMMMMMNMMNDOI=~NM?      =MMM+   NMMMO     8M$   ,MM,  $MMMN      =D,      ,?N~     , ?NMMMNNNDDO8DDD887,             ,$ND= :O$NMZ  IMO                                                                                     ,IMMMMN$, 7NO,                 ,,,,,:,,,
+#                                                                                                                 ,~IDMNN8OI,       :~+$DMMMMMMMN87+=~~~~?ZDNNMM,      ?NM?   :NMN     :MM7   ~MN ~DMMMN        O~     :DD,         ,MMMMNNOI:,,       ,     ,=ZODD8D?    :MMM:  NMZ                                     +8NM8,                                           ,,       :~                      ,,~:, 
+#                                                                                                                         ,INNMMNNO+           :=78NNZIIONMNNDMMN       ,7D~           :MN~    MNMM?=MM          $?   OD,         ,IMD     ,:8MNMNNNNNNNNMMNM7:     :8~  ZND    OMN7,                                 :NNMMMMN                                                                       :8MMMMMNZ=: 
+#                                                                                                                              :O88NMMN$=~:,           ?MMN88NMMMD                     =MN?     =,  :NM           +7$NO,          =N8 ,       OMI, ,,     7M:        ZDDND?     MMO         ,,                       +MM8?OMN:                                                                       ,::         
+#                                                                                                                                     :$NMMMNMM8I:     ,8MD,   +MMN=            ,=?77=:ZMMM7        ?MN             ~$+          7M7         ,DZ          :M~       IMM8:     ,8MN,               , ,,,,           :MMD,=DMM+,?O8= ,  ,                                                                          
+#                                                                                                                                          ,=I77$ONNNZ?+ONMZ     =$D          ,+=::+$8NNMNDNMD7?,   ?MN                :~~:     ~D7          ,NI          ,M7   ~?ZNZ?,       8MM~                               =ZMMN~$MMMMMMMNMMD$ONMMMMD:                                                                     
+#                                                                                                                                                        ~DMM=                   ~I7=,       =DMMMD7MMM,                   :+?=?O$:          ~N:          ~MN88D$:           NMM=,                              ~MMMMMMMMMMMMOZMMMMMM7::8MMI                                                                     
+#                                                                                                                                                        , :DNNI                                :~IDMMND                        :::=?II?==~::ZN7=+I$ZZZ8DZ+~~:             IMMM~                               ~MMMNMMMMMI~:7NMMMMD7,: +NM8,                                                                     
+#                                                                                                                                                            :NMM7,                                  ~MM+                               ,,,:~==~~~:  ,                    OMMN:                               ?NNMMMMM8,   ~NMNO,     =MMMN?,,                                                                   
+#                                                                                   ,,,,,,,,                                                                    ,$8MMO=                                ~M8                                                              +MNO:                                ,MM~ ,        ::          ,~,          ,,:::,,,,                                                    
+#                                                                          ,::,,,,                                                                                  ~ONND+                               OD+                                                          ?NMD,                                   ??                                            ,::::::,,                                            
+#                                                             :=+??=:,,,,,                                                                                              ~?$D87I:                          ~Z?                                                      =$DM$:                                                                                           ,::,,,,,,,,                                  
+#                                                       ,:~==~,   :+=,                                                                                                       $NNNNZ~             ,,:~~:    ,                                                    :DMMMI                                                                                                                                           
+#                                              ,~~~~~:,,,             ,,,                                                                                                       ,~IMMMN8O+,             ,:~?7$Z7~,                                          :ZDMNNDNM8ZI                                                                                                                                         
+#                                      ,,~~:,,                            ,                                                                                                          :$DNNMMN8?, ,            ,~7ZOO?:                                 ,:$NMMMM?     :7NMMD?,                                                                                                                                    
+#                              ,:~~:,,,                                                                                                                                                      $NMMMMMNMNO?~             ~?ODDD$=,                :?8MMMMMD?~7NMMMD$~        :ONMMN?                                                                                                                               
+#                     ,,,,,,,,,                                                                                                                                                                 ?$DMMMMMMMMMMD$?=~,        ,~7ZZODN87????I$ODNMDOZ$7I:         ~$ZDMD7==~      =$ZDND$++~,                                                                                                                       
+#          ~?++=~~,                                                                                                                                                                                    ~+?II7ZNMMMMMM8$$$?~,           ,~?II7Z8DDOZ$77II+:            ~?IZDM8O$I~,   :??$8MMNZZ7+,                                                                                                               
+#            ,                                                                           , ,+D7                                                                                                                   :=IZ8NMMMNNMNNO$=               ,:?ZDNNMMMNMND8Z7=,       =ZNMMMMMD?,,    :ONMNMMMN?,                                                                                                          
+#                                                                                         ,MMMM7                                                                                                                             ,,::~IONNMMNNDDD8OZI=,         ,::::=+I$ONMNNNNDDDNNMMMMMMMMMD87:   ,:~=ZNNNDOI,            ~7$ZO8DDNNNNDD8O+:,                                                                     
+#~,                                                                                       +MMMMDI?~                                                                                                                                       ,~+Z8DNDNNMMMMDD$+:,,, ,           ,~?7O8DNNNNNNMMMMMNOOZI??++?IZ88$:  ,,~ZDMMMMMMMMMMMMMMMMMMNNNNNMMMNND8$=,,                                                         
+# ,~:                                                                                     NMMMMOZMM8:      ,7DZ~                                                                                                                                        ,,~IONMMMMMMMMMNDZ+~:               ,     ,=I8DNMMMMMNMNMMMMMDNMMMMNMMMNNNDDDDD8O8Z$7II+++IZDDMMNMN$,,                                                   
+#   :~:                                                                                   MMMMMNMMM+ :, +?DOI~                                                                                                                                                     ,:~?7$$ZODDNMNN8Z7??+=~:,                 ,~=+?7I?=:,,,,:=?I7$$$$$$$ZZZOO8DDNNMMNNNNNMMMMNMZ+                                                 
+#     ~=,                                                                                 MMMMMMMM8   ,NNNO                                         ,                                                                                                                             ,~?ZDNMNNNNNNND8O7?:,,,                                          ,~=7DMMMMMMMMMZ~                                              
+#      ,:+~                                                                              ,MMMMMMMMO  ~N8: ,,:~,                            ,:~:,,,                                                                                        ,:~,                                              ,:~~=+++?7$O8DNNNNNMMN8$=,           ~+$ZO8DNNMMNNNND8OOOZ$+:, :=+I8MMMMDOI                                          
+#         ,=+                                                                             MMMMMMMMMDMZ,         ~?:               ,,,,,                                                                                                         ,:~~~,                                                          ,NN?$NMMMMMMMMMMMN87~:,,,=+=:,,,,,,:?NMMNMMMMMMNNDNDI8MMM+  ,                                    
+#            +=,                                                                          $MMMN?$NMN,            ,+?+:,    ,,:~~:,                                                                                                                    :=~,                                                      8M=     ,                :7ONNNO?~,   , :$NNMMMMMMMMMMMMMN,                                      
+#             ,+?                                                                         ,NNO,,  ,                 :ODNNNZ:                                                                                                                                :?777I=                                             IMO           ,,=$Z7+~        ,?NNMMNZ:       :7NMMMMMNMMMM8                         , ,IDND~    
+#               ,=~,                                                                        7I                :+I+~::,, ~?I~                                                                                                                              , ,~=~==:                                              ZMZ      ?8I:     :I$DND$?~      :?$8MMN$?:      ?ZDMMMMMMMM7                      +$NMMMMMI    
+#                  ~~                                                                      ,             ,=+=:              ,==                                                                                                                       ::,               ,                                         ZMM:     ?NN8:        ,7NMMNI,       ,$NMMMO:       =NMMMNMM+                     I8NMMMM,     
+#                    ~=                                                                          ,~:,,~~,                      ,:,                                                                                                              ,,,,                        , ,,                                   +MN8     ,~OMM8Z,        :+ZMNDOI       =IMMN8=      ,=NNMO                      ?NMMMD       
+#                      ::,                                                                  ,,,::                                  ,:,                                                                                                       ,,                                                                      =MMO~    , ?NMMMI,         ,INMMMDI,     ~DMMN+,      NMO                      ?DNO~        
+#                        ,~,                                                      ,,,::,                                                ,,                                                                                           ,                                                   ,,                            ,INMND+:    :MMM+              ~ZDDMMMD?~:$MMMMMMMMMMM?                                   
+#                           ,,                                             :~~:,                                                                                                                                               ,:,                                                             ,,                          :ZMMMMNNMMMN,                   ,=ONMMMMNM$,,,,,,                                   ,,
+#Z,                           :,                                   ,, ,~=~,                                                                                                                                                ,,                                                                                                  ,=?7$I= ,                        :~~~,                                      ,,,,  
+#MD=                            ::                             ,,,,                                                                                                                                                  , ,,                                                                                                                                                                              ,,,,      
+#MNMI                             ::                                                                                                                                                                            ,,,                                                                                                                                                                                 ,,,,         
+#7MMM?         ,                   ,~:                                                                                                                                                                     :~:,                                                                                                                                                                                  ,:,             
+# OMMMO:     ~NMD,                    :=:                                                                                                                                                            :+=:                                                                                                                                                                                    ,,,                 
+# ~MMMN8:    NMMM~                      +?:                                                                                                                                                    ,:=++~                                                                                                                                                                                    ,:,,                    
+#  $MMMMMNI,?MMMMZ ,DM7                   ,=I=                                                                                                                                           ~+~~:,                                                                                                                                                                                     ,:,,,                        
+#  +MMO~OMMMMMMMMD MMM7 , ,                  =$~                                                                                                                                     ?OZ+                                                                                                                                                                                         ,:,                            
+#  ~NMN: :+DNN7MMMNMMMDODMMN+                  :+?,                                                                                                                            +77+~:                                                                                                                                                                                         :::,                               
+#   IMM+      ,NMMMMMMMMN?NM7                    ~7+,   ,                                                                                                                  :+$I,                                                            ,  +8?                                                                                                                         :+~                                   
+#   =NMD,      NMMMMMM8~ ?NN,                      =ND?                                                                                                              ,=??:                                                                 ,8NMMMM=                                                                                                                    :++                                       
+#    +DO,      ~ZZ$ODI  :8M~                  :=I8NMNMMD+,                                                                                                      ,:~~=:                                                                 , +8MO$DMD+   ~===~:                                                                                                         :~~:                                         
+#                       :$~                 7NMD$,   ZMM8I?                                                                                                 ,~=:                                                                     , ZNN7,:MM8~,OMNMD8DMM=                                                                                                     ++:                                             
+#                                        =ZNOI:    ?DND=  ,?I~                                                                                          ,,,:                                                                        ,$ND+ ?NMNNNNN7+,   7MN,                                                                                                 ,=+~                                                
+#                                      :ZI:        :,        ,=7:                                                                                ::,,                                                                             $NN= 7NMMMM7:      :DMMMN8NMMDDMN7                                                                                     ,::,                                                    
+#                                                               ~?=                                                                          ,,,                                                                                ,ZM=  DMMD8         ?MMMMNN7,    ,I+                                                                                  :~~,                                                       
+#                                                                  =+,                                                                  ,,,                                                                                           ,,            =Z8:                                                                                           +$~                                                           
+#                                                                    :~                                                                                                                                                                                                                                                                        ,,~~,                                                             
+#                                                                        :,                                                                                                                                                                                                                                                                  ::,                                                                 
+#                                                                         ::,                                                                                                                                                                                                                                                            ,:::,                                                                   
+#                                                                            ,,                                                                                                                                                                                                                                                        ,~                                                                        
+#                                                                               ,,                                                                                                                                                                                                                                                 ,,,                                                                           
+#                                                                                                                                                                                                                                                                                                                                                                                                
+#
\ No newline at end of file
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