File: ReformatPacBio.java

package info (click to toggle)
bbmap 39.01%2Bdfsg-2
  • links: PTS, VCS
  • area: main
  • in suites: bookworm
  • size: 21,760 kB
  • sloc: java: 267,418; sh: 15,163; python: 5,247; ansic: 2,074; perl: 96; xml: 38; makefile: 38
file content (1438 lines) | stat: -rwxr-xr-x 46,126 bytes parent folder | download | duplicates (2)
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
241
242
243
244
245
246
247
248
249
250
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
274
275
276
277
278
279
280
281
282
283
284
285
286
287
288
289
290
291
292
293
294
295
296
297
298
299
300
301
302
303
304
305
306
307
308
309
310
311
312
313
314
315
316
317
318
319
320
321
322
323
324
325
326
327
328
329
330
331
332
333
334
335
336
337
338
339
340
341
342
343
344
345
346
347
348
349
350
351
352
353
354
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
372
373
374
375
376
377
378
379
380
381
382
383
384
385
386
387
388
389
390
391
392
393
394
395
396
397
398
399
400
401
402
403
404
405
406
407
408
409
410
411
412
413
414
415
416
417
418
419
420
421
422
423
424
425
426
427
428
429
430
431
432
433
434
435
436
437
438
439
440
441
442
443
444
445
446
447
448
449
450
451
452
453
454
455
456
457
458
459
460
461
462
463
464
465
466
467
468
469
470
471
472
473
474
475
476
477
478
479
480
481
482
483
484
485
486
487
488
489
490
491
492
493
494
495
496
497
498
499
500
501
502
503
504
505
506
507
508
509
510
511
512
513
514
515
516
517
518
519
520
521
522
523
524
525
526
527
528
529
530
531
532
533
534
535
536
537
538
539
540
541
542
543
544
545
546
547
548
549
550
551
552
553
554
555
556
557
558
559
560
561
562
563
564
565
566
567
568
569
570
571
572
573
574
575
576
577
578
579
580
581
582
583
584
585
586
587
588
589
590
591
592
593
594
595
596
597
598
599
600
601
602
603
604
605
606
607
608
609
610
611
612
613
614
615
616
617
618
619
620
621
622
623
624
625
626
627
628
629
630
631
632
633
634
635
636
637
638
639
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
655
656
657
658
659
660
661
662
663
664
665
666
667
668
669
670
671
672
673
674
675
676
677
678
679
680
681
682
683
684
685
686
687
688
689
690
691
692
693
694
695
696
697
698
699
700
701
702
703
704
705
706
707
708
709
710
711
712
713
714
715
716
717
718
719
720
721
722
723
724
725
726
727
728
729
730
731
732
733
734
735
736
737
738
739
740
741
742
743
744
745
746
747
748
749
750
751
752
753
754
755
756
757
758
759
760
761
762
763
764
765
766
767
768
769
770
771
772
773
774
775
776
777
778
779
780
781
782
783
784
785
786
787
788
789
790
791
792
793
794
795
796
797
798
799
800
801
802
803
804
805
806
807
808
809
810
811
812
813
814
815
816
817
818
819
820
821
822
823
824
825
826
827
828
829
830
831
832
833
834
835
836
837
838
839
840
841
842
843
844
845
846
847
848
849
850
851
852
853
854
855
856
857
858
859
860
861
862
863
864
865
866
867
868
869
870
871
872
873
874
875
876
877
878
879
880
881
882
883
884
885
886
887
888
889
890
891
892
893
894
895
896
897
898
899
900
901
902
903
904
905
906
907
908
909
910
911
912
913
914
915
916
917
918
919
920
921
922
923
924
925
926
927
928
929
930
931
932
933
934
935
936
937
938
939
940
941
942
943
944
945
946
947
948
949
950
951
952
953
954
955
956
957
958
959
960
961
962
963
964
965
966
967
968
969
970
971
972
973
974
975
976
977
978
979
980
981
982
983
984
985
986
987
988
989
990
991
992
993
994
995
996
997
998
999
1000
1001
1002
1003
1004
1005
1006
1007
1008
1009
1010
1011
1012
1013
1014
1015
1016
1017
1018
1019
1020
1021
1022
1023
1024
1025
1026
1027
1028
1029
1030
1031
1032
1033
1034
1035
1036
1037
1038
1039
1040
1041
1042
1043
1044
1045
1046
1047
1048
1049
1050
1051
1052
1053
1054
1055
1056
1057
1058
1059
1060
1061
1062
1063
1064
1065
1066
1067
1068
1069
1070
1071
1072
1073
1074
1075
1076
1077
1078
1079
1080
1081
1082
1083
1084
1085
1086
1087
1088
1089
1090
1091
1092
1093
1094
1095
1096
1097
1098
1099
1100
1101
1102
1103
1104
1105
1106
1107
1108
1109
1110
1111
1112
1113
1114
1115
1116
1117
1118
1119
1120
1121
1122
1123
1124
1125
1126
1127
1128
1129
1130
1131
1132
1133
1134
1135
1136
1137
1138
1139
1140
1141
1142
1143
1144
1145
1146
1147
1148
1149
1150
1151
1152
1153
1154
1155
1156
1157
1158
1159
1160
1161
1162
1163
1164
1165
1166
1167
1168
1169
1170
1171
1172
1173
1174
1175
1176
1177
1178
1179
1180
1181
1182
1183
1184
1185
1186
1187
1188
1189
1190
1191
1192
1193
1194
1195
1196
1197
1198
1199
1200
1201
1202
1203
1204
1205
1206
1207
1208
1209
1210
1211
1212
1213
1214
1215
1216
1217
1218
1219
1220
1221
1222
1223
1224
1225
1226
1227
1228
1229
1230
1231
1232
1233
1234
1235
1236
1237
1238
1239
1240
1241
1242
1243
1244
1245
1246
1247
1248
1249
1250
1251
1252
1253
1254
1255
1256
1257
1258
1259
1260
1261
1262
1263
1264
1265
1266
1267
1268
1269
1270
1271
1272
1273
1274
1275
1276
1277
1278
1279
1280
1281
1282
1283
1284
1285
1286
1287
1288
1289
1290
1291
1292
1293
1294
1295
1296
1297
1298
1299
1300
1301
1302
1303
1304
1305
1306
1307
1308
1309
1310
1311
1312
1313
1314
1315
1316
1317
1318
1319
1320
1321
1322
1323
1324
1325
1326
1327
1328
1329
1330
1331
1332
1333
1334
1335
1336
1337
1338
1339
1340
1341
1342
1343
1344
1345
1346
1347
1348
1349
1350
1351
1352
1353
1354
1355
1356
1357
1358
1359
1360
1361
1362
1363
1364
1365
1366
1367
1368
1369
1370
1371
1372
1373
1374
1375
1376
1377
1378
1379
1380
1381
1382
1383
1384
1385
1386
1387
1388
1389
1390
1391
1392
1393
1394
1395
1396
1397
1398
1399
1400
1401
1402
1403
1404
1405
1406
1407
1408
1409
1410
1411
1412
1413
1414
1415
1416
1417
1418
1419
1420
1421
1422
1423
1424
1425
1426
1427
1428
1429
1430
1431
1432
1433
1434
1435
1436
1437
1438
package icecream;

import java.io.PrintStream;
import java.util.ArrayList;
import java.util.Random;

import aligner.Aligner;
import consensus.BaseGraph;
import dna.AminoAcid;
import dna.Data;
import fileIO.ByteFile;
import fileIO.ByteStreamWriter;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import json.JsonObject;
import prok.GeneCaller;
import shared.KillSwitch;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.ReadStats;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import shared.TrimRead;
import stream.ConcurrentReadOutputStream;
import stream.FASTQ;
import stream.FastaReadInputStream;
import stream.Read;
import stream.SamLine;
import structures.ByteBuilder;
import structures.EntropyTracker;
import structures.IntHashSet;
import structures.IntList;

/**
 * Version of Reformat designed for PacBio data.
 * Supports some of Reformat's capability, like subsampling,
 * in a ZMW-aware tool.
 * 
 * @author Brian Bushnell
 * @date June 5, 2019
 *
 */
public final class ReformatPacBio {
	
	/*--------------------------------------------------------------*/
	/*----------------        Initialization        ----------------*/
	/*--------------------------------------------------------------*/
	
	/**
	 * Code entrance from the command line.
	 * @param args Command line arguments
	 */
	public static void main(String[] args){
		//Start a timer immediately upon code entrance.
		Timer t=new Timer();
		
		//Create an instance of this class
		ReformatPacBio x=new ReformatPacBio(args);
		
		//Run the object
		x.process(t);
		
		//Close the print stream if it was redirected
		Shared.closeStream(x.outstream);
	}
	
	/**
	 * Constructor.
	 * @param args Command line arguments
	 */
	public ReformatPacBio(String[] args){
		
		{//Preparse block for help, config files, and outstream
			PreParser pp=new PreParser(args, getClass(), false);
			args=pp.args;
			outstream=pp.outstream;
		}
		
		//Set shared static variables prior to parsing
		ReadWrite.USE_PIGZ=ReadWrite.USE_UNPIGZ=true;
		ReadWrite.USE_BGZIP=ReadWrite.USE_UNBGZIP=ReadWrite.PREFER_BGZIP=true;
		ReadWrite.MAX_ZIP_THREADS=Shared.threads();
		FASTQ.TEST_INTERLEAVED=FASTQ.FORCE_INTERLEAVED=false;
		SamLine.SET_FROM_OK=true;
		Shared.setBufferData(1000000);
//		Shared.FASTA_WRAP=511;
		Data.USE_SAMBAMBA=false;//Sambamba changes PacBio headers.
		Read.CHANGE_QUALITY=false;
		EntropyTracker.defaultK=3;
		
		{//Parse the arguments
			final Parser parser=parse(args);
			Parser.processQuality();
			
			maxReads=parser.maxReads;
			overwrite=ReadStats.overwrite=parser.overwrite;
			append=ReadStats.append=parser.append;
			
			in1=parser.in1;
			extin=parser.extin;

			if(outg==null){outg=parser.out1;}
			extout=parser.extout;
		}
		
		sampleReadsExact=sampleReadsTarget>-1;
		sampleBasesExact=sampleBasesTarget>-1;
		sampleZMWsExact=sampleZMWsTarget>-1;
		sampleExact=(sampleReadsExact || sampleBasesExact || sampleZMWsExact);
		
		//Determine how many threads may be used
		threads=(sampleExact || (samplerate>0 && seed>=0)) ? 1 : Shared.threads();
		
		//Ensure there is an input file
		if(in1==null){throw new RuntimeException("Error - at least one input file is required.");}
		fixExtensions(); //Add or remove .gz or .bz2 as needed
		checkFileExistence(); //Ensure files can be read and written
		checkStatics(); //Adjust file-related static fields as needed for this program 
		
		//Create output FileFormat objects
		ffoutg=FileFormat.testOutput(outg, FileFormat.FASTQ, extout, true, overwrite, append, false);
		ffoutb=FileFormat.testOutput(outb, FileFormat.FASTQ, extout, true, overwrite, append, false);
		ffstats=FileFormat.testOutput(outstats, FileFormat.TXT, null, false, overwrite, append, false);
		ffschist=FileFormat.testOutput(schist, FileFormat.TXT, null, false, overwrite, append, false);
		
		//Create input FileFormat objects
		ffin1=FileFormat.testInput(in1, FileFormat.FASTQ, extin, true, true);
		if(verbose){System.err.println("Finished constructor for "+getClass().getName());}
	}
	
	/*--------------------------------------------------------------*/
	/*----------------    Initialization Helpers    ----------------*/
	/*--------------------------------------------------------------*/
	
	/** Parse arguments from the command line */
	private Parser parse(String[] args){
		//Create a parser object
		Parser parser=new Parser();
		
		//Set any necessary Parser defaults here
		//parser.foo=bar;
		
		//Parse each argument
		for(int i=0; i<args.length; i++){
			String arg=args[i];
			
			//Break arguments into their constituent parts, in the form of "a=b"
			String[] split=arg.split("=");
			String a=split[0].toLowerCase();
			String b=split.length>1 ? split[1] : null;
			if(b!=null && b.equalsIgnoreCase("null")){b=null;}
			
			if(a.equals("verbose")){
				verbose=Parse.parseBoolean(b);
			}else if(a.equals("format")){
				if(b==null){
					assert(false) : arg;
				}else if(Tools.isDigit(b.charAt(i))){
					format=Integer.parseInt(b);
				}else if(b.equalsIgnoreCase("json")){
					format=FORMAT_JSON;
				}else if(b.equalsIgnoreCase("text")){
					format=FORMAT_TEXT;
				}else{
					assert(false) : arg;
				}
				assert(format>=1 && format<=2) : arg;
			}else if(a.equals("json")){
				boolean x=Parse.parseBoolean(b);
				format=(x ? FORMAT_JSON : FORMAT_TEXT);
			}else if(a.equals("ss") || a.equals("samstreamer") || a.equals("streamer")){
				if(b!=null && Tools.isDigit(b.charAt(0))){
					ZMWStreamer.useStreamer=true;
					assert(Integer.parseInt(b)==1) : "ZMWStreamer threads currently capped at 1.";
//					ZMWStreamer.streamerThreads=Tools.max(1, Integer.parseInt(b));
				}else{
					ZMWStreamer.useStreamer=Parse.parseBoolean(b);
				}
			}
			
			else if(a.equals("keepshortreads") || a.equals("ksr")){
				keepShortReads=Parse.parseBoolean(b);
			}else if(a.equalsIgnoreCase("keepzmwstogether") || a.equals("kzt") || a.equals("keepreadstogether") || a.equals("krt")){
				keepZMWsTogether=Parse.parseBoolean(b);
			}else if(a.equalsIgnoreCase("subsampleFromEnds")){
				subsampleFromEnds=Parse.parseBoolean(b);
			}
			
			else if(a.equals("ccsinput") || a.equals("ccsin")){
				CCSInput=Parse.parseBoolean(b);
			}else if(a.equals("minlength") || a.equals("minlen")){
				minLengthAfterTrimming=Integer.parseInt(b);
			}else if(a.equals("flaglongreads")){
				flagLongReads=Parse.parseBoolean(b);
			}else if(a.equals("longreadmult")){
				longReadMult=Float.parseFloat(b);
			}else if(a.equals("trimreads") || a.equals("trim")){
				trimReads=Parse.parseBoolean(b);
			}else if(a.equals("parsecustom")){
				parseCustom=Parse.parseBoolean(b);
			}else if(a.equals("outg") || a.equals("outgood")){
				outg=b;
			}else if(a.equals("outb") || a.equals("outbad")){
				outb=b;
			}else if(a.equals("outs") || a.equals("outstats") || a.equals("stats")){
				outstats=b;
			}else if(a.equals("schist") || a.equals("shist")){
				schist=b;
			}
			
			else if(a.equals("shredlen")){
				shredLength=Integer.parseInt(b);
			}else if(a.equals("overlap")){
				overlap=Integer.parseInt(b);
			}else if(a.equalsIgnoreCase("minShredIdentity") || a.equalsIgnoreCase("minShredId") ||
					 a.equalsIgnoreCase("shredId")){
				minShredIdentity=Tools.max(0.01f, Float.parseFloat(b));
				if(minShredIdentity>1){minShredIdentity*=0.01f;}
				assert(minShredIdentity>0 && minShredIdentity<=1) : 
					"minShredIdentity ranges from 0 (exclusive) to 1 (inclusive).";
			}else if(a.equals("ccs") || a.equals("makeccs") || a.equals("consensus")){
				makeCCS=Parse.parseBoolean(b);
			}else if(a.equals("findorientation") || a.equals("orient") || a.equals("reorient")){
				findOrientation=Parse.parseBoolean(b);
			}
			
			else if(a.equals("minpasses")){
				minPasses=Float.parseFloat(b);
			}else if(a.equals("minsubreads")){
				minSubreads=Integer.parseInt(b);
			}
			
			else if(a.equalsIgnoreCase("samplerate")){
				samplerate=Float.parseFloat(b);
			}else if(a.equalsIgnoreCase("sampleReadsTarget") || a.equals("srt")){
				sampleReadsTarget=Parse.parseKMG(b);
			}else if(a.equalsIgnoreCase("sampleBasesTarget") || a.equals("sbt")){
				sampleBasesTarget=Parse.parseKMG(b);
			}else if(a.equalsIgnoreCase("sampleZMWsTarget") || a.equals("szt")){
				sampleZMWsTarget=Parse.parseKMG(b);
			}else if(a.equals("bestpass") || a.equals("keepbestpass") || 
					a.equals("keepbest") || a.equals("bestpassonly")){
				keepBestPass=Parse.parseBoolean(b);
			}else if(a.equals("longestpass") || a.equals("keeplongestpass") || 
					a.equals("keeplongest") || a.equals("longestpassonly")){
				keepLongestPass=Parse.parseBoolean(b);
			}else if(a.equals("seed")){
				seed=Long.parseLong(b);
			}
			
			else if(a.equalsIgnoreCase("zmws") || a.equalsIgnoreCase("maxzmws")){
				maxZMWs=Parse.parseKMG(b);
			}
			
			else if(a.equals("whitelist")){
				whitelist=b;
			}else if(a.equals("whitelist")){
				blacklist=b;
			}
			
			else if(a.equalsIgnoreCase("trimpolya")){
				trimPolyA=Parse.parseBoolean(b);
			}else if(PolymerTrimmer.parse(arg, a, b)){
				//do nothing
			}else if(a.equals("minentropy") || a.equals("entropy") || a.equals("entropyfilter") || a.equals("efilter")){
				if(b==null || Character.isLetter(b.charAt(0))){
					if(Parse.parseBoolean(b)){
						entropyCutoff=0.55f;
					}else{
						entropyCutoff=-1;
					}
				}else{
					entropyCutoff=Float.parseFloat(b);
				}
			}else if(a.equals("entropyblock") || a.equals("entropylength") || a.equals("entropylen") || a.equals("entlen")){
				entropyLength=Parse.parseIntKMG(b);
			}else if(a.equals("entropyfraction") || a.equals("entfraction")){
				entropyFraction=Float.parseFloat(b);
			}else if(a.equals("monomerfraction") || a.equals("maxmonomerfraction") || a.equals("mmf")){
				maxMonomerFraction=Float.parseFloat(b);
			}else if(a.equals("parse_flag_goes_here")){
				long fake_variable=Parse.parseKMG(b);
				//Set a variable here
			}else if(parser.parse(arg, a, b)){//Parse standard flags in the parser
				//do nothing
			}else{
				outstream.println("Unknown parameter "+args[i]);
				assert(false) : "Unknown parameter "+args[i];
			}
		}

		if(verbose){System.err.println("Finished parser for "+getClass().getName());}
		return parser;
	}
	
	/** Add or remove .gz or .bz2 as needed */
	private void fixExtensions(){
		in1=Tools.fixExtension(in1);
	}
	
	/** Ensure files can be read and written */
	private void checkFileExistence(){
		
		//Ensure output files can be written
		if(!Tools.testOutputFiles(overwrite, append, false, outg, outb, outstats, schist)){
			outstream.println((outg==null)+", "+(outb==null)+", "+outg+", "+outb+", "+outstats);
			throw new RuntimeException("\n\noverwrite="+overwrite+"; Can't write to output files "+
					outg+", "+outb+", "+outstats+", "+schist+"\n");
		}
		
		//Ensure input files can be read
		if(!Tools.testInputFiles(false, true, in1, whitelist, blacklist)){
			throw new RuntimeException("\nCan't read some input files.\n");  
		}
		
		//Ensure that no file was specified multiple times
		if(!Tools.testForDuplicateFiles(true, in1, outg, outb, outstats, schist, whitelist, blacklist)){
			throw new RuntimeException("\nSome file names were specified multiple times.\n");
		}
	}
	
	/** Adjust file-related static fields as needed for this program */
	private static void checkStatics(){
		//Adjust the number of threads for input file reading
		if(!ByteFile.FORCE_MODE_BF1 && !ByteFile.FORCE_MODE_BF2 && Shared.threads()>2){
			ByteFile.FORCE_MODE_BF2=true;
		}
		
		assert(FastaReadInputStream.settingsOK());
	}
	
	/*--------------------------------------------------------------*/
	/*----------------         Outer Methods        ----------------*/
	/*--------------------------------------------------------------*/
	
	/** Create read streams and process all data */
	void process(Timer t){
		if(verbose){System.err.println("Entered process()");}

		whiteSet=Tools.loadIntSet(whitelist);
		blackSet=Tools.loadIntSet(blacklist);

		//Count reads in the original file
		if(sampleExact){countInitial();}
		
		//Turn off read validation in the input threads to increase speed
		final boolean vic=Read.VALIDATE_IN_CONSTRUCTOR;
		Read.VALIDATE_IN_CONSTRUCTOR=Shared.threads()<4;
		
		//Create a read input stream
		ZMWStreamer zstream=new ZMWStreamer(ffin1, Shared.threads(), maxReads, maxZMWs);
		
		//Optionally create read output streams
		final ConcurrentReadOutputStream rosg=makeCros(ffoutg);
		final ConcurrentReadOutputStream rosb=makeCros(ffoutb);
		
		//Reset counters
		readsProcessed=readsOut=0;
		basesProcessed=basesOut=0;
		
		//Process the reads in separate threads
		spawnThreads(zstream, rosg, rosb);
		
		if(verbose){outstream.println("Finished; closing streams.");}
		
		//Write anything that was accumulated by ReadStats
		errorState|=ReadStats.writeAll();
		//Close the read streams
		errorState|=ReadWrite.closeStreams(null, rosg, rosb);
		
		//Reset read validation
		Read.VALIDATE_IN_CONSTRUCTOR=vic;
		
		writeHistogram(ffschist, subreadCounts);
		
		//Report timing and results
		t.stop();
		
		String stats=null;
		if(format==FORMAT_TEXT){
			ByteBuilder bb=toText(t);
			stats=bb.nl().toString();
		}else if(format==FORMAT_JSON){
			JsonObject jo=toJson(t);
			stats=jo.toStringln();
		}else{
			assert(false) : "Bad format: "+format;
		}
		if(ffstats==null){
			outstream.print(stats);
		}else{
			ReadWrite.writeString(stats, outstats);
		}
		
		//Throw an exception of there was an error in a thread
		if(errorState){
			throw new RuntimeException(getClass().getName()+" terminated in an error state; the output may be corrupt.");
		}
	}
	
	private ByteBuilder toText(Timer t){
		ByteBuilder bb=new ByteBuilder();
		bb.appendln(Tools.timeZMWsReadsBasesProcessed(t, ZMWs, readsProcessed, basesProcessed, 8));
		bb.appendln(Tools.ZMWsReadsBasesOut(ZMWs, readsProcessed, basesProcessed, ZMWsOut, readsOut, basesOut, 8, true));
		
		long readsFiltered=readsProcessed-readsOut;
		bb.appendln(Tools.numberPercent("Reads Filtered:", readsFiltered, readsFiltered*100.0/(readsProcessed), 3, 8));
		if(trimReads || trimPolyA){
			bb.appendln(Tools.numberPercent("Reads Trimmed:", readsTrimmed, readsTrimmed*100.0/(readsProcessed), 3, 8));
			bb.appendln(Tools.numberPercent("Bases Trimmed:", basesTrimmed, basesTrimmed*100.0/(basesProcessed), 3, 8));
		}
//		bb.appendln(Tools.number("Total ZMWs:", ZMWs, 8));
		bb.appendln(Tools.numberPercent("Partial ZMWs:", partiallyDiscardedZMWs, partiallyDiscardedZMWs*100.0/(ZMWs), 3, 8));
		bb.appendln(Tools.numberPercent("Discarded ZMWs:", fullyDiscardedZMWs, fullyDiscardedZMWs*100.0/(ZMWs), 3, 8));
//		bb.appendln(Tools.numberPercent("Low Entropy:", lowEntropyReads, lowEntropyReads*100.0/(readsProcessed), 3, 8));
		if(entropyCutoff>=0){
			bb.appendln(Tools.numberPercent("Low Entropy:", lowEntropyZMWs, lowEntropyZMWs*100.0/(ZMWs), 3, 8));
		}
		
		if(parseCustom){}
		return bb;
	}
	
	private JsonObject toJson(Timer t){
		JsonObject jo=new JsonObject();
		long readsFiltered=readsProcessed-readsOut;
		
//		asdf
		jo.add("Time", t.timeInSeconds());
		jo.add("ZMWs_Processed", ZMWs);
		jo.add("Reads_Processed", readsProcessed);
		jo.add("Bases_Processed", basesProcessed);
		jo.add("Reads_Out", readsOut);
		jo.add("Bases_Out", basesOut);
		jo.add("ZMWs_Out", ZMWsOut);
		jo.add("Reads_Filtered", readsFiltered);
		jo.add("Reads_Filtered_Pct", readsFiltered*100.0/(readsProcessed));
		if(trimReads){
			jo.add("Reads_Trimmed", readsTrimmed);
			jo.add("Reads_Trimmed_Pct", readsTrimmed*100.0/(readsProcessed));
			jo.add("Bases_Trimmed", basesTrimmed);
			jo.add("Bases_Trimmed_Pct", basesTrimmed*100.0/(basesProcessed));
		}
//		jo.add("Total_ZMWs", ZMWs);fullyDiscardedZMWs
		jo.add("Partial_ZMWs", partiallyDiscardedZMWs);
		jo.add("Partial_ZMWs_Pct", partiallyDiscardedZMWs*100.0/(ZMWs));
		jo.add("Discarded_ZMWs", fullyDiscardedZMWs);
		jo.add("Discarded_ZMWs_Pct", fullyDiscardedZMWs*100.0/(ZMWs));
//		jo.add("Low_Entropy", lowEntropyReads);
//		jo.add("Low_Entropy_Pct", lowEntropyReads*100.0/(readsProcessed));
		if(entropyCutoff>=0){
			jo.add("Low_Entropy", lowEntropyZMWs);
			jo.add("Low_Entropy_Pct", lowEntropyZMWs*100.0/(ZMWs));
		}
		if(parseCustom){
			//Special stats if desired
		}
		return jo;
	}
	
	private static void writeHistogram(FileFormat ff, long[] hist){
		if(ff==null){return;}
		final ByteStreamWriter bsw=new ByteStreamWriter(ff);
		bsw.start();

		bsw.print("#Counted\t").println(Tools.sum(hist));
		bsw.print("#Mean\t").println(Tools.averageHistogram(hist), 3);
		bsw.print("#Median\t").println(Tools.medianHistogram(hist));
		bsw.print("#Mode\t").println(Tools.calcModeHistogram(hist));
		bsw.print("#STDev\t").println(Tools.standardDeviationHistogram(hist), 3);
		bsw.print("#Value\tOccurances\n");
		
		for(int i=0; i<hist.length; i++){
			bsw.print(i).tab().println(hist[i]);
		}
		bsw.poisonAndWait();
	}
	
	private ConcurrentReadOutputStream makeCros(FileFormat ff){
		if(ff==null){return null;}

		//Select output buffer size based on whether it needs to be ordered
		final int buff=16;

		final ConcurrentReadOutputStream ros=ConcurrentReadOutputStream.getStream(
				ff, null, buff, null, ff.samOrBam() && ffin1.samOrBam());
		ros.start(); //Start the stream
		return ros;
	}
	
	/** Spawn process threads */
	private void spawnThreads(final ZMWStreamer zstream,
			final ConcurrentReadOutputStream rosg,
			final ConcurrentReadOutputStream rosb){
		
		//Do anything necessary prior to processing
		
		//Fill a list with ProcessThreads
		ArrayList<ProcessThread> alpt=new ArrayList<ProcessThread>(threads);
		for(int i=0; i<threads; i++){
			alpt.add(new ProcessThread(zstream, rosg, rosb, i));
		}
		
		//Start the threads
		for(ProcessThread pt : alpt){
			pt.start();
		}
		
		zstream.runStreamer(false);
		
		//Wait for threads to finish
		waitForThreads(alpt);
		
		//Do anything necessary after processing
		ZMWs=zstream.ZMWs;
	}
	
	private void waitForThreads(ArrayList<ProcessThread> alpt){
		
		//Wait for completion of all threads
		boolean success=true;
		for(ProcessThread pt : alpt){
			
			//Wait until this thread has terminated
			while(pt.getState()!=Thread.State.TERMINATED){
				try {
					//Attempt a join operation
					pt.join();
				} catch (InterruptedException e) {
					//Potentially handle this, if it is expected to occur
					e.printStackTrace();
				}
			}
			
			//Accumulate per-thread statistics
			readsProcessed+=pt.readsProcessedT;
			basesProcessed+=pt.basesProcessedT;
			ZMWs+=pt.ZMWsT;
			
			readsOut+=pt.readsOutT;
			basesOut+=pt.basesOutT;
			ZMWsOut+=pt.ZMWsOutT;

			partiallyDiscardedZMWs+=pt.partiallyDiscardedZMWsT;
			fullyDiscardedZMWs+=pt.fullyDiscardedZMWsT;
			lowEntropyZMWs+=pt.lowEntropyZMWsT;
			lowEntropyReads+=pt.lowEntropyReadsT;
			
			basesTrimmed+=pt.basesTrimmedT;
			readsTrimmed+=pt.readsTrimmedT;
			
			success&=pt.success;
		}
		
		//Track whether any threads failed
		if(!success){errorState=true;}
	}
	
	/*--------------------------------------------------------------*/
	/*----------------          CCS Methods         ----------------*/
	/*--------------------------------------------------------------*/
	
	public Read makeConsensus(ZMW zmw){
		//Apply correct strand
		for(int i=0; i<zmw.size(); i++){
			zmw.get(i).setStrand(i&1);
		}
		final Read ref=zmw.medianRead(false);
		if(zmw.size()<3 || zmw.estimatePasses()<2.1){return ref;}
		
		BaseGraph bg=new BaseGraph(ref.id, ref.bases, ref.quality, ref.numericID, 0);
		float idSum=0;
		int added=0;
		for(Read r : zmw){
			if(r!=ref){
				final boolean rcomp=(r.strand()!=ref.strand());
				if(rcomp){r.reverseComplement();}
				float id=shredAndAdd(r, bg, null);
				idSum+=id;
				added++;
				if(rcomp){r.reverseComplement();}
			}
		}
		float avgId=idSum/(Tools.max(1, added));
		
		//TODO: Also interesting to get a traversal quality here...
		Read c=bg.traverse();
		c.obj=avgId;
		
		//If not enough subreads aligned, discard the result
		if(minPasses>1 && bg.baseTotal/(float)ref.length()<minPasses-1){c.setDiscarded(true);}
		return c;
	}
	
	private float shredAndAdd(Read subread, BaseGraph bg, Aligner ssa){
		int added=0;
		float idSum=0;
		if(ssa==null){ssa=GeneCaller.getSSA();}
		if(subread.length()<=shredLength){
			float id=bg.alignAndGenerateMatch(subread, ssa, findOrientation, minShredIdentity);
			added++;
			idSum+=id;
			if(id>=minShredIdentity){
				bg.add(subread);
			}
		}else{
			ArrayList<Read> shreds=shred(subread, shredLength, overlap);
			for(int i=0; i<shreds.size(); i++){
				Read shred=shreds.get(i);
				float id=bg.alignAndGenerateMatch(shred, ssa, findOrientation, minShredIdentity);
				added++;
				idSum+=id;
				if(id>=minShredIdentity){
					bg.add(shred, (i>0 ? overlap-1 : 0), 0);
				}
			}
		}
		float avgId=idSum/(Tools.max(1, added));
		return avgId;
	}
	
	private static ArrayList<Read> shred(final Read r, final int shredLength, final int overlap){
		final byte[] bases=r.bases;
		final byte[] quals=r.quality;
		final String name=r.id;
		final int increment=shredLength-overlap;
		final double incMult=1.0f/increment;
		final int chunks=bases.length<=shredLength ? 1 : (int)Math.ceil((bases.length-overlap)*incMult);
		assert(chunks>0);
		final double inc2=bases.length/(double)chunks;
		
		final ArrayList<Read> list=new ArrayList<Read>(chunks);
		if(chunks==1){
			list.add(r);
			return list;
		}
		
		for(int chunk=0; chunk<chunks; chunk++){
			int a=(int)Math.floor(inc2*chunk);
			int b=(chunk==chunks-1 ? bases.length : overlap+(int)Math.floor(inc2*(chunk+1)));
			b=Tools.min(b, a+shredLength);
			final int length=b-a;
//			if(length<minLength){return;}
			final byte[] bases2=KillSwitch.copyOfRange(bases, a, b);
			final byte[] quals2=(quals==null ? null : KillSwitch.copyOfRange(quals, a, b));
			Read shred=new Read(bases2, quals2, name+"_"+a+"-"+(b-1), r.numericID);
//			readsOut++;
//			basesOut+=shred.length();
			list.add(shred);
		}
		return list;
	}
	
//	/** Generates match string and returns identity */
//	public float alignAndGenerateMatch(Read r, Aligner ssa){
//		if(ssa==null){ssa=GeneCaller.getSSA();}
//		byte[] query=r.bases;
//		int a=0, b=ref.length-1;
//		int[] max=ssa.fillUnlimited(query, original, a, b, -9999);
//		if(max==null){return 0;}
//		
//		final int rows=max[0];
//		final int maxCol=max[1];
//		final int maxState=max[2];
//		final byte[] match=ssa.traceback(query, original, a, b, rows, maxCol, maxState);
//		int[] score=ssa.score(query, original, a, b, rows, maxCol, 0);
//		r.match=match;
//		r.start=score[1];
//		r.stop=score[2];
//		r.setMapped(true);
//		final float identity=Read.identity(match);
//		return identity;
//	}
	
	/*--------------------------------------------------------------*/
	/*----------------         Inner Methods        ----------------*/
	/*--------------------------------------------------------------*/
	
	void countInitial(){
		if(verbose){System.err.println("Entered countInitial()");}
		assert(!ffin1.stdio()) : "Target subsampling can't be used with stdin.";
		initialReads=0;
		initialZMWs=0;
		initialBases=0;
		ZMWStreamer zstream=new ZMWStreamer(ffin1, Shared.threads(), maxReads, maxZMWs);
		zstream.runStreamer(true);
		for(ZMW zmw=zstream.nextZMW(); zmw!=null; zmw=zstream.nextZMW()){
			initialZMWs++;
			initialReads+=zmw.size();
			initialBases+=zmw.countBases();
		}
		readsRemaining=initialReads;
		ZMWsRemaining=initialZMWs;
		basesRemaining=initialBases;
		if(verbose){System.err.println("Finished countInitial()");}
	}
	
	/*--------------------------------------------------------------*/
	/*----------------         Inner Classes        ----------------*/
	/*--------------------------------------------------------------*/
	
	/** Processes reads. */
	private class ProcessThread extends Thread {
		
		//Constructor
		ProcessThread(final ZMWStreamer zstream_, 
				final ConcurrentReadOutputStream ros_,  
				final ConcurrentReadOutputStream rosb_, final int tid_){
			zstream=zstream_;
			rosg=ros_;
			rosb=rosb_;
			tid=tid_;
			
			if(entropyCutoff>=0){
				eTracker=new EntropyTracker(false, entropyCutoff, true);
			}else{
				eTracker=null;
			}
		}
		
		//Called by start()
		@Override
		public void run(){
			//Do anything necessary prior to processing
			randy=Shared.threadLocalRandom(seed<0 ? seed : seed+tid);
			//Randy can't be initialized in the constructor because it is threadlocal;
			//that leads to nonrandom results.
			
			//Process the reads
			processInner();
			
			//Do anything necessary after processing
			
			//Indicate successful exit status
			success=true;
		}
		
		/** Iterate through the reads */
		void processInner(){

			//As long as there is a nonempty read list...
			for(ZMW reads=zstream.nextZMW(); reads!=null; reads=zstream.nextZMW()){
//				if(verbose){outstream.println("Fetched "+reads.size()+" reads.");} //Disabled due to non-static access
				
				processList(reads);
			}
		}
		
		private int medianLength(ZMW list){
			if(list.size()<3){return -1;}
			IntList lengths=new IntList(list.size()-2);
			
			for(int i=1; i<list.size()-1; i++){
				lengths.add(list.get(i).length());
			}
			lengths.sort();
			int median=lengths.get((lengths.size-1)/2);
			return median;
		}
		
		int flagLowEntropyReads(final ZMW reads, final float minEnt, 
				final int minLen0, final float minFract){
			int found=0;
			for(Read r : reads){
				if(!r.discarded()){
					int minLen=Tools.min((int)(r.length()*minFract), minLen0);
					int maxBlock=eTracker.longestLowEntropyBlock(r.bases, false, maxMonomerFraction);
					if(maxBlock>=minLen){
						r.setDiscarded(true);
						r.setJunk(true);
						found++;
//						System.err.println(r.toFasta());
					}
				}
			}
			return found;
		}
		
		int flagLongReads(final ZMW reads, int median){
			int found=0;
			for(Read r : reads){
				if(r.length()>longReadMult*median){
					r.setDiscarded(true);
					r.setHasAdapter(true);
					found++;
				}
			}
			return found;
		}
		
		void discardEndReads(ZMW zmw, int toDiscard){
			if(toDiscard<1){return;}
			if(toDiscard==1){
				if(zmw.first().length()<zmw.last().length()){
					zmw.first().setDiscarded(true);
				}else{
					zmw.last().setDiscarded(true);
				}
				return;
			}else{
				zmw.first().setDiscarded(true);
				zmw.last().setDiscarded(true);
				for(int i=2; i<toDiscard; i++){
					zmw.get(zmw.size()-i).setDiscarded(true);
				}
			}
		}
		
		/** Each list is presumed to be all reads from a ZMW, in order */
		void processList(final ZMW reads){
			ZMWsT++;
			{
				int idx=Tools.min(subreadCounts.length-1, reads.size());
				synchronized(subreadCounts){
					//TODO: Slow
					//But other statistics could be gathered here as well
					subreadCounts[idx]++;
				}
			}
			long numBases=0;
			
			//Loop through each read in the list for statistics
			for(int idx=0; idx<reads.size(); idx++){
				final Read r1=reads.get(idx);
				
				//Validate reads in worker threads
				if(!r1.validated()){r1.validate(true);}

				//Track the initial length for statistics
				final int initialLength1=r1.length();

				//Increment counters
				readsProcessedT++;
				basesProcessedT+=initialLength1;
				numBases+=initialLength1;
			}
			final boolean fullPass=CCSInput || reads.size()>=3;

			if(whiteSet!=null || blackSet!=null){
				int zid=reads.zid();
				assert(zid>=0) : "Can't parse ZMW ID from header "+reads.get(0).id;
				if(whiteSet!=null && !whiteSet.contains(zid)){reads.setDiscarded(true);}
				if(blackSet!=null && blackSet.contains(zid)){reads.setDiscarded(true);}
			}

			final int subreads=reads.size();
			final float passes=reads.estimatePasses();
			if((minPasses>0 && passes<minPasses) || (minSubreads>0 && reads.size()<minSubreads)){
				reads.setDiscarded(true);
			}
			
			if(samplerate<1){
				if(keepZMWsTogether){
					if(randy.nextFloat()>samplerate){
						for(Read r : reads){r.setDiscarded(true);}
					}
				}else if(subsampleFromEnds){
					//Roll a fraction of the reads
					int remove=0;
					for(Read r : reads){
						if(randy.nextFloat()>samplerate){
							remove++;
						}
					}
					discardEndReads(reads, remove);
				}else{
					for(Read r : reads){
						if(randy.nextFloat()>samplerate){
							r.setDiscarded(true);
						}
					}
				}
			}
			
			if(keepBestPass || keepLongestPass){
				Read best=keepBestPass ? reads.medianRead(false) : reads.longestRead(false);
//				assert(reads.size()<5) : keepBestPass+", "+keepLongestPass+
//					", "+best.length()+", "+Arrays.toString(reads.lengths());
				for(Read r : reads){
					if(r!=best){
						r.setDiscarded(true);
					}
				}
			}else if(makeCCS && !reads.discarded()){
				Read r=makeConsensus(reads);
				reads.clear();
				reads.add(r);
				//TODO: Fix read header so that it looks like a CCS read.
				PBHeader pbh=new PBHeader(r.id);
				float avgId=(r.obj==null ? 0 : (Float)r.obj);
				r.id=pbh.runID+"/"+pbh.zmwID+"/ccs"
						+ "\tsubreads="+subreads+"\tpasses="+String.format("%.2f", passes)
						+"\tavgID="+String.format("%.4f", avgId);
				
//				ZMW.fixReadHeader(r, 0, trimmed);
			}
			
			if(sampleExact){
				if(sampleReadsExact){
					if(keepZMWsTogether){
						assert(readsRemaining>0) : readsRemaining;
						double prob=sampleReadsTarget/(double)(readsRemaining);
						if(randy.nextDouble()<prob){sampleReadsTarget-=reads.size();}
						else{reads.setDiscarded(true);}
						readsRemaining-=reads.size();
					}else{
						for(Read r : reads){
							if(r!=null && !r.discarded()){
								assert(readsRemaining>0) : readsRemaining;
								double prob=sampleReadsTarget/(double)(readsRemaining);
								if(randy.nextDouble()<prob){sampleReadsTarget--;}
								else{r.setDiscarded(true);}
							}
							readsRemaining--;
						}
					}
				}else if(sampleBasesExact){
					if(keepZMWsTogether){
						assert(basesRemaining>0) : basesRemaining;
						final long bases=reads.countBases();
						double prob=sampleBasesTarget/(double)(basesRemaining);
						if(randy.nextDouble()<prob){sampleBasesTarget-=bases;}
						else{reads.setDiscarded(true);}
						basesRemaining-=bases;
					}else{
						for(Read r : reads){
							if(r!=null && !r.discarded()){
								assert(basesRemaining>0) : basesRemaining;
								final int bases=r.length();
								double prob=sampleBasesTarget/(double)(basesRemaining);
								if(randy.nextDouble()<prob){sampleBasesTarget-=bases;}
								else{r.setDiscarded(true);}
								basesRemaining-=bases;
							}
						}
					}
				}else if(sampleZMWsExact){
					assert(ZMWsRemaining>0) : ZMWsRemaining;
					double prob=sampleZMWsTarget/(double)(ZMWsRemaining);
					if(randy.nextDouble()<prob){sampleZMWsTarget--;}
					else{reads.setDiscarded(true);}
					ZMWsRemaining--;
				}else{assert(false) : "No sampling mode.";}
			}
			
			if(trimReads || trimPolyA){
				int removed=0;
				for(int i=0; i<reads.size(); i++){
					Read r=reads.get(i);
					if(!r.discarded()){
						byte a=r.bases[0], b=r.bases[r.length()-1];
						int trimmed=0;
						if(!AminoAcid.isFullyDefined(a) || !AminoAcid.isFullyDefined(b)){
							trimmed+=trimRead(r, 80);
						}
						if(trimPolyA){
							int x=trimPolyA(r);
							trimmed+=x;
							if(x>0){r.setTrimmed(true);}
						}

						if(trimmed>0){
							basesTrimmedT+=trimmed;
							readsTrimmedT++;
							//TODO: technically this should track the lefty and right trim amounts
							//but it doesn't really matter as long as the sum is correct
							ZMW.fixReadHeader(r, 0, trimmed);
						}

						//TODO: Note again, removing intermediate reads messes up forward-rev ordering
						if(r.length()<minLengthAfterTrimming){//Discard short trash
							//						reads.set(i, null);
							r.setDiscarded(true);
							removed++;
						}
					}
				}
			}
			
			if(entropyCutoff>0){
				int bad=flagLowEntropyReads(reads, entropyCutoff, entropyLength, entropyFraction);
				if(bad>0){
					lowEntropyZMWsT++;
					lowEntropyReadsT+=bad;
					if(bad>=reads.size()){
						if(!reads.isEmpty()){outputReads(reads);}
						return; //No point in continuing...
					}
				}
			}
			
			if(reads.isEmpty()){return;}

			Read sample=null;//Read that will be searched for inverted repeat, typically median length
			Read shortest=null;//shortest read in the middle, or overall if no middle reads
			final int medianLength=medianLength(reads);
			int longReads=0;
			int sampleNum=0;
			
			if(reads.size()>=3){
				if(flagLongReads){longReads=flagLongReads(reads, medianLength);}
				for(int i=1; i<reads.size()-1; i++){
					Read r=reads.get(i);
					if(sample==null && r.length()==medianLength){
						sample=r;
						sampleNum=i;
					}
					if(shortest==null || r.length()<shortest.length()){shortest=r;}
				}
			}else{
				for(int i=0; i<reads.size(); i++){
					Read r=reads.get(i);
					if(sample==null || sample.length()<r.length()){
						sample=r;
						sampleNum=i;
					}
					if(shortest==null || r.length()<shortest.length()){shortest=r;}
				}
			}
			assert(sample!=null);
			
			if(keepShortReads){
				
			}else if(sample.discarded() || (longReads>0)){
				for(Read r : reads){
					r.setDiscarded(true);
				}
			}
			
			if(minLengthAfterTrimming>0){removeShortTrash(reads);}
			
			if(!reads.isEmpty()){outputReads(reads);}
		}
		
		private int removeShortTrash(ZMW reads) {
			int removed=0;
			for(int i=0; i<reads.size(); i++){
				Read r=reads.get(i);
				if(r.length()<minLengthAfterTrimming){//Discard short trash
					if(!r.discarded()){
						removed++;
						r.setDiscarded(true);
					}
				}
			}
			return removed;
		}
		
		private void fixReadHeader(Read r, int leftTrim, int rightTrim){
			leftTrim=Tools.max(0, leftTrim);
			rightTrim=Tools.max(0, rightTrim);
			if(leftTrim<1 && rightTrim<1){return;}
			final int idx=r.id.lastIndexOf('/');
			if(idx>0 && idx<r.id.length()-3){
				String prefix=r.id.substring(0, idx+1);
				String suffix=r.id.substring(idx+1);
				if(suffix.indexOf('_')>0){
					String coords=suffix, comment="";
					int tab=suffix.indexOf('\t');
					if(tab<0){tab=suffix.indexOf(' ');}
					if(tab>0){
						coords=coords.substring(0, tab);
						comment=coords.substring(tab);
					}
					String[] split=Tools.underscorePattern.split(coords);
					int left=Integer.parseInt(split[0]);
					int right=Integer.parseInt(split[1]);
					left+=leftTrim;
					right-=rightTrim;
					if(left>right){left=right;}
					
					if(right-left!=r.length()){right=left+r.length();}
//					System.err.println(r.length()+", "+(right-left));
					
					r.id=prefix+left+"_"+right+comment;
					final SamLine sl=r.samline;
					if(sl!=null){
						sl.qname=r.id;
						if(sl.optional!=null){
							for(int i=0; i<sl.optional.size(); i++){
								String s=sl.optional.get(i);
								if(s.startsWith("qe:i:")){
									s="qe:i:"+right;
									sl.optional.set(i, s);
								}else if(s.startsWith("qs:i:")){
									s="qs:i:"+left;
									sl.optional.set(i, s);
								}
							}
						}
					}
				}
			}
		}
		
		int trimRead(Read r, int lookahead){
			final byte[] bases=r.bases;
			
			int left=calcLeftTrim(bases, lookahead);
			int right=calcRightTrim(bases, lookahead);
			int trimmed=0;
			
			if(left+right>0){
//				System.err.println(r.length()+", "+left+", "+right+", "+r.id);
				trimmed=TrimRead.trimByAmount(r, left, right, 1, false);
//				System.err.println(r.length()+", "+left+", "+right+", "+r.id);
				fixReadHeader(r, left, right);
//				System.err.println(r.length()+", "+left+", "+right+", "+r.id);
			}
			
			if(r.samline!=null){
				r.samline.seq=r.bases;
				r.samline.qual=r.quality;
			}
			return trimmed;
		}
		
		int trimPolyA(Read r){
			final byte[] bases=r.bases;

			int left=Tools.max(PolymerTrimmer.testLeft(bases, 'A'), PolymerTrimmer.testLeft(bases, 'T'));
			int right=Tools.max(PolymerTrimmer.testRight(bases, 'A'), PolymerTrimmer.testRight(bases, 'T'));
			int trimmed=0;
			
			if(left+right>0){
//				System.err.println(r.length()+", "+left+", "+right+", "+r.id);
				trimmed=TrimRead.trimByAmount(r, left, right, 1, false);
//				System.err.println(r.length()+", "+left+", "+right+", "+r.id);
				fixReadHeader(r, left, right);
//				System.err.println(r.length()+", "+left+", "+right+", "+r.id);
			}
			
			if(r.samline!=null){
				r.samline.seq=r.bases;
				r.samline.qual=r.quality;
			}
			return trimmed;
		}
		
		final int calcLeftTrim(final byte[] bases, int lookahead){
			final int len=bases.length;
			int lastUndef=-1;
			for(int i=0, defined=0; i<len && defined<lookahead; i++){
				if(AminoAcid.isFullyDefined(bases[i])){
					defined++;
				}else{
					lastUndef=i;
					defined=0;
				}
			}
			return lastUndef+1;
		}
		
		final int calcRightTrim(final byte[] bases, int lookahead){
			final int len=bases.length;
			int lastUndef=len;
			for(int i=len-1, defined=0; i>=0 && defined<lookahead; i--){
				if(AminoAcid.isFullyDefined(bases[i])){
					defined++;
				}else{
					lastUndef=i;
					defined=0;
				}
			}
			return len-lastUndef-1;
		}
		
		private void outputReads(ZMW reads){
			final int size=reads.size();
			final ArrayList<Read> good=new ArrayList<Read>(size);
			final ArrayList<Read> bad=new ArrayList<Read>(size);
//			final ArrayList<Read> bad=(rosb==null ? null : new ArrayList<Read>(size));
			
			int discardedReads=0;
			int trimmedReads=0;
			boolean sendAllToDiscarded=false;
			
			//Check to see if any reads are discarded or ambiguous
			for(Read r : reads){
				if(r.discarded()){
					discardedReads++;
				}else if(r.trimmed()){
					trimmedReads++;
				}
			}
			
			//Unify flags on all reads
			if(keepZMWsTogether){
				if(discardedReads>0){sendAllToDiscarded=true;}
			}
			if(discardedReads>0){
				if(discardedReads==size){
					fullyDiscardedZMWsT++;
				}else{
					partiallyDiscardedZMWsT++;
				}
			}
			
			for(Read r : reads){
				if(r.discarded() || sendAllToDiscarded){
//					assert(false);
					if(bad!=null){bad.add(r);}
//					System.err.println("d:t="+r.tested()+",ad="+r.hasAdapter()+",ir="+r.invertedRepeat()+","+r.id+", reads="+reads.size());
				}else{
					if(good!=null){
						good.add(r);
					}
					readsOutT++;
					basesOutT+=r.length();
				}
			}

			if(good!=null && !good.isEmpty()){ZMWsOutT++;}
			if(rosg!=null && good!=null && !good.isEmpty()){rosg.add(good, 0);}
			if(rosb!=null && bad!=null && !bad.isEmpty()){rosb.add(bad, 0);}
		}
		
		/*--------------------------------------------------------------*/
		/*----------------      Inner Class Fields      ----------------*/
		/*--------------------------------------------------------------*/
		
		/** Number of reads processed by this thread */
		protected long readsProcessedT=0;
		/** Number of bases processed by this thread */
		protected long basesProcessedT=0;
		/** Number of ZMWs processed by this thread */
		protected long ZMWsT=0;
		
		protected long basesTrimmedT=0;
		protected long readsTrimmedT=0;
		
		/** Number of reads retained by this thread */
		protected long readsOutT=0;
		/** Number of bases retained by this thread */
		protected long basesOutT=0;
		/** Number of ZMWs retained by this thread */
		protected long ZMWsOutT=0;

		protected long partiallyDiscardedZMWsT=0;
		protected long fullyDiscardedZMWsT=0;
		
		protected long lowEntropyZMWsT=0;
		protected long lowEntropyReadsT=0;
		
		/** True only if this thread has completed successfully */
		boolean success=false;
		
		/** Shared output stream */
		private final ConcurrentReadOutputStream rosg;
		/** Shared output stream for bad reads */
		private final ConcurrentReadOutputStream rosb;
		/** Thread ID */
		final int tid;
		
		final ZMWStreamer zstream;
		final EntropyTracker eTracker;
		Random randy;
	}
	
	/*--------------------------------------------------------------*/
	/*----------------            Fields            ----------------*/
	/*--------------------------------------------------------------*/
	
	/** Primary input file path */
	private String in1=null;

	/** Primary output file path */
	private String outg=null;
	/** Bad output file path */
	private String outb=null;
	
	/** Stats (screen) output file path */
	private String outstats=null;
	
	/** Subread count histogram */
	private String schist=null;
	
	/** Override input file extension */
	private String extin=null;
	/** Override output file extension */
	private String extout=null;
	
	private float longReadMult=1.5f;
	
	/** For grading synthetic data */
	private boolean parseCustom;
	
	/** Input reads are CCS (full pass) */
	private boolean CCSInput;
	
	//Note: These flags are very similar... they need to be better-defined or merged.
	private boolean keepZMWsTogether=false;
	private boolean keepShortReads=true;
	
	private boolean subsampleFromEnds=false;
	
	private int format=FORMAT_TEXT;
	private static final int FORMAT_TEXT=1, FORMAT_JSON=2;
	
	/*--------------------------------------------------------------*/
	
	/** Number of reads processed */
	protected long readsProcessed=0;
	/** Number of bases processed */
	protected long basesProcessed=0;

	/** Number of reads retained */
	protected long readsOut=0;
	/** Number of bases retained */
	protected long basesOut=0;
	/** Number of ZMWs retained */
	protected long ZMWsOut=0;

	protected long partiallyDiscardedZMWs=0;
	protected long fullyDiscardedZMWs=0;
	
	/** Quit after processing this many input reads; -1 means no limit */
	private long maxReads=-1;
	
	protected long basesTrimmed=0;
	protected long readsTrimmed=0;
	
	protected long lowEntropyZMWs=0;
	protected long lowEntropyReads=0;
	
	/** Total ZMWs observed */
	protected long ZMWs=0;
	
	/** Histogram */
	protected final long[] subreadCounts=new long[101];
	
	private boolean flagLongReads=false;
	private boolean trimReads=false;
	private int minLengthAfterTrimming=0;
	
	boolean trimPolyA=false;
	
	/*--------------------------------------------------------------*/
	/*----------------      Subsampling Fields      ----------------*/
	/*--------------------------------------------------------------*/
	
	//If a target is chosen, these will be initialized with the original counts
	long initialReads=0;
	long initialZMWs=0;
	long initialBases=0;
	
	long readsRemaining=0;
	long ZMWsRemaining=0;
	long basesRemaining=0;
	
	float samplerate=1.0f;
	long sampleReadsTarget=-1;
	long sampleBasesTarget=-1;
	long sampleZMWsTarget=-1;

	final boolean sampleReadsExact;
	final boolean sampleBasesExact;
	final boolean sampleZMWsExact;
	final boolean sampleExact;
	
	boolean keepBestPass=false;
	boolean keepLongestPass=false;
	
	/*--------------------------------------------------------------*/
	/*----------------        Subset Fields         ----------------*/
	/*--------------------------------------------------------------*/

	String whitelist;
	String blacklist;
	
	IntHashSet whiteSet;
	IntHashSet blackSet;
	
	/*--------------------------------------------------------------*/
	/*----------------        Entropy Fields        ----------------*/
	/*--------------------------------------------------------------*/
	
	/** Minimum entropy to be considered "complex", on a scale of 0-1 */
	float entropyCutoff=-1; //suggested 0.55
	/** Minimum length of a low-entropy block to fail a read */
	int entropyLength=350;
	/** Minimum length of a low-entropy block as a fraction of read length;
	 * the minimum of the two will be used */
	float entropyFraction=0.5f;
	
	float maxMonomerFraction=0.74f; //Suggested...  0.74
	
	/*--------------------------------------------------------------*/
	/*----------------         Final Fields         ----------------*/
	/*--------------------------------------------------------------*/

	/** Primary input file */
	private final FileFormat ffin1;
	
	/** Primary output file */
	private final FileFormat ffoutg;
	
	/** Bad output file */
	private final FileFormat ffoutb;

	/** Stats output file */
	private final FileFormat ffstats;

	/** Subread count histogram */
	private final FileFormat ffschist;
	
	private final int threads;
	
	private long seed=-1;
	private long maxZMWs=-1;
	
	private int shredLength=500;
	private int overlap=10;//Helps make the shreds concur at their borders.
	private float minShredIdentity=0.6f;
	private boolean makeCCS=false;
	private boolean findOrientation=false;
	
	private float minPasses=0;
	private int minSubreads=0;
	
	/*--------------------------------------------------------------*/
	/*----------------        Common Fields         ----------------*/
	/*--------------------------------------------------------------*/
	
	/** Print status messages to this output stream */
	private PrintStream outstream=System.err;
	/** Print verbose messages */
	public static boolean verbose=false;
	/** True if an error was encountered */
	public boolean errorState=false;
	/** Overwrite existing output files */
	private boolean overwrite=false;
	/** Append to existing output files */
	private boolean append=false;
	
}