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package jgi;
import java.io.File;
import java.io.PrintStream;
import java.util.ArrayList;
import java.util.Collections;
import dna.AminoAcid;
import fileIO.ByteFile;
import fileIO.ByteFile1;
import fileIO.ByteFile2;
import fileIO.ByteStreamWriter;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import shared.TrimRead;
import stream.ConcurrentGenericReadInputStream;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FASTQ;
import stream.FastaReadInputStream;
import stream.Read;
import stream.SamLine;
import structures.ByteBuilder;
import structures.ListNum;
import structures.LongPair;
/**
* @author Brian Bushnell
* @date Oct 17, 2014
*
*/
public class FindHiCJunctions {
public static void main(String[] args){
//Start a timer immediately upon code entrance.
Timer t=new Timer();
//Create an instance of this class
FindHiCJunctions x=new FindHiCJunctions(args);
//Run the object
x.process(t);
//Close the print stream if it was redirected
Shared.closeStream(x.outstream);
}
public FindHiCJunctions(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
Shared.capBuffers(4); //Only for singlethreaded programs
ReadWrite.USE_PIGZ=ReadWrite.USE_UNPIGZ=true;
ReadWrite.setZipThreads(Shared.threads());
FASTQ.TEST_INTERLEAVED=FASTQ.FORCE_INTERLEAVED=false;
Parser parser=new Parser();
for(int i=0; i<args.length; i++){
String arg=args[i];
String[] split=arg.split("=");
String a=split[0].toLowerCase();
String b=split.length>1 ? split[1] : null;
if(a.equals("verbose")){
verbose=Parse.parseBoolean(b);
ByteFile1.verbose=verbose;
ByteFile2.verbose=verbose;
stream.FastaReadInputStream.verbose=verbose;
ConcurrentGenericReadInputStream.verbose=verbose;
stream.FastqReadInputStream.verbose=verbose;
ReadWrite.verbose=verbose;
}else if(a.equals("minclip")){
minClipLength=Integer.parseInt(b);
}else if(a.equals("printkmers") || a.equals("printcounts")){
printKmers=Parse.parseBoolean(b);
}else if(a.equals("junctionfile") || a.equals("junctions") || a.equals("outk")){
junctionFile=b;
assert(b==null || b.indexOf('%')>0);
}else if(parser.parse(arg, a, b)){
//do nothing
}
else if(parser.in1==null && i==0 && Tools.looksLikeInputStream(arg)){
parser.in1=arg;
}else if(parser.out1==null && i==1 && !arg.contains("=")){
parser.out1=arg;
}else{
outstream.println("Unknown parameter "+args[i]);
assert(false) : "Unknown parameter "+args[i];
// throw new RuntimeException("Unknown parameter "+args[i]);
}
}
{//Process parser fields
Parser.processQuality();
maxReads=parser.maxReads;
overwrite=parser.overwrite;
append=parser.append;
in1=parser.in1;
out1=parser.out1;
extin=parser.extin;
extout=parser.extout;
}
assert(FastaReadInputStream.settingsOK());
if(in1==null){throw new RuntimeException("Error - at least one input file is required.");}
if(!ByteFile.FORCE_MODE_BF1 && !ByteFile.FORCE_MODE_BF2){
ByteFile.FORCE_MODE_BF2=false;
ByteFile.FORCE_MODE_BF1=true;
}
if(out1!=null && out1.equalsIgnoreCase("null")){out1=null;}
if(!Tools.testOutputFiles(overwrite, append, false, out1)){
outstream.println((out1==null)+", "+out1);
throw new RuntimeException("\n\noverwrite="+overwrite+"; Can't write to output files "+out1+"\n");
}
ffout1=FileFormat.testOutput(out1, FileFormat.FASTQ, extout, true, overwrite, append, false);
ffin1=FileFormat.testInput(in1, FileFormat.FASTQ, extin, true, true);
counts=new long[11][];
counts[10]=new long[1024*1024];
counts[8]=new long[256*256];
counts[6]=new long[4096];
counts[4]=new long[256];
leftCounts=new long[6][];
rightCounts=new long[6][];
leftCounts[5]=new long[1024];
leftCounts[4]=new long[256];
leftCounts[3]=new long[64];
leftCounts[2]=new long[16];
rightCounts[5]=new long[1024];
rightCounts[4]=new long[256];
rightCounts[3]=new long[64];
rightCounts[2]=new long[16];
}
void process(Timer t){
final ConcurrentReadInputStream cris;
{
cris=ConcurrentReadInputStream.getReadInputStream(maxReads, true, ffin1, null, null, null);
cris.start();
if(verbose){outstream.println("Started cris");}
}
final ConcurrentReadOutputStream ros;
if(out1!=null){
final int buff=4;
assert(!out1.equalsIgnoreCase(in1) && !out1.equalsIgnoreCase(in1)) : "Input file and output file have same name.";
ros=ConcurrentReadOutputStream.getStream(ffout1, null, null, null, buff, null, false);
ros.start();
}else{ros=null;}
long readsProcessed=0, readsOut=0;
long basesProcessed=0, basesOut=0;
{
ListNum<Read> ln=cris.nextList();
ArrayList<Read> reads=(ln!=null ? ln.list : null);
// outstream.println("Fetched "+reads);
if(reads!=null && !reads.isEmpty()){
Read r=reads.get(0);
assert((ffin1==null || ffin1.samOrBam()) || (r.mate!=null)==cris.paired());
}
while(ln!=null && reads!=null && reads.size()>0){//ln!=null prevents a compiler potential null access warning
final ArrayList<Read> listOut=new ArrayList<Read>(reads.size());
for(int idx=0; idx<reads.size(); idx++){
final Read r1=reads.get(idx);
final int initialLength1=r1.length();
final boolean keep=process(r1, r1.samline);
if(keep || true){
listOut.add(r1);
readsOut++;
basesOut+=r1.length();
}
readsProcessed++;
basesProcessed+=initialLength1;
}
if(ros!=null){ros.add(listOut, ln.id);}
cris.returnList(ln);
ln=cris.nextList();
reads=(ln!=null ? ln.list : null);
}
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
errorState|=ReadWrite.closeStreams(cris, ros);
if(printKmers){
printKmers(10, counts[10], "");
printKmers(8, counts[8], "");
printKmers(6, counts[6], "");
printKmers(4, counts[4], "");
printKmers(5, leftCounts[5], "L");
printKmers(4, leftCounts[4], "L");
printKmers(3, leftCounts[3], "L");
printKmers(5, rightCounts[5], "R");
printKmers(4, rightCounts[4], "R");
printKmers(3, rightCounts[3], "R");
}
t.stop();
outstream.println(Tools.timeReadsBasesProcessed(t, readsProcessed, basesProcessed, 8));
outstream.println(Tools.readsBasesOut(readsProcessed, basesProcessed, readsOut, basesOut, 8, false));
if(errorState){
throw new RuntimeException(getClass().getName()+" terminated in an error state; the output may be corrupt.");
}
}
private void printKmers(int k, long[] array, String direction){
boolean tsv=junctionFile.endsWith(".tsv") || junctionFile.endsWith(".tsv.gz");
final String fname=junctionFile.replaceFirst("%", k+direction);
final ByteStreamWriter bsw=new ByteStreamWriter(fname, true, false, false);
bsw.start();
final long sum=shared.Vector.sum(array);
final double mult=1.0/(Tools.max(1, sum));
final long thresh=Tools.max((2*sum)/array.length, (long)Math.ceil(Tools.max(minCount, minFraction*sum)));
// if(k==4){System.err.println(Arrays.toString(array));}
//
//// assert(k!=4) : thresh+", "+sum;
ArrayList<LongPair> list=new ArrayList<LongPair>();
for(int kmer=0; kmer<array.length; kmer++){
final long count=array[kmer];
if(count>=thresh){
list.add(new LongPair(count, kmer));
}
}
Collections.sort(list);
Collections.reverse(list);
ByteBuilder bb=new ByteBuilder(4200);
for(LongPair pair : list){
if(tsv){
bb.append(AminoAcid.kmerToString(pair.b, k)).append('\t');
bb.append(pair.a).append('\t').append(mult*pair.a, 5).append('\n');
}else{
bb.append('>').append(pair.a).append('\t').append(mult*pair.a, 5).append('\n');
bb.append(AminoAcid.kmerToString(pair.b, k)).append('\n');
}
if(bb.length()>4096){
bsw.print(bb);
bb.clear();
}
}
if(!bb.isEmpty()){
bsw.print(bb);
}
errorState|=bsw.poisonAndWait();
}
/*--------------------------------------------------------------*/
private boolean process(Read r, SamLine sl){
if(sl==null || !sl.mapped() || !sl.primary() || sl.supplementary()|| r.match==null || !r.containsNonNM()){
return false;
}
boolean definiteJunction=(sl.nextMapped() && !sl.properPair());
// final String cigar=sl.cigar;
final byte[] bases=r.bases;
final boolean rcomp=(r.strand()==Shared.MINUS);
final int left, right;
{
if(r.shortmatch()){r.toLongMatchString(true);}
int left0=SamLine.countLeadingClip(r.match);
int right0=SamLine.countTrailingClip(r.match);
if(left0==0 && right0==0){
byte[] smatch=softClipMatch(r.match, minClipLength, true);
left0=SamLine.countLeadingClip(smatch);
right0=SamLine.countTrailingClip(smatch);
}
left=left0;
right=right0;
}
if((left>1 && right>1) || (left<minClipLength && right<minClipLength)){return false;}
final int pos=(left>right ? left-1 : bases.length-right-1);
if(printKmers && definiteJunction && junctionFile!=null){
if(rcomp){r.reverseComplement();}
//Pos: position of base to left of junction
// System.err.println(left+", "+right+", "+pos+", "+rcomp);
// System.err.println(new String(bases).substring(0, pos+1)+"~"+new String(bases).substring(pos+1));
for(int k=10, half=5, start=pos-4; k>=4; k-=2, half--, start++){
int kmer=0;
assert(start>=0) : left+", "+right+", "+pos+", "+start+", "+k+", "+bases.length+"\n"+new String(r.match);
for(int i=start, len=0; len<k; i++, len++){
final byte b=bases[i];
final int num=AminoAcid.baseToNumber[b];
kmer=(kmer<<2)|num;
}
if(kmer<0){return false;}
// System.err.println("Adding "+AminoAcid.kmerToString(kmer, k));
counts[k][kmer]++;
leftCounts[half][kmer>>k]++;
rightCounts[half][kmer&~((-1)<<k)]++;
}
if(rcomp){r.reverseComplement();}
}
if(trim){
int trimAmount=bases.length-pos-1;
int remainingAmount=bases.length-trimAmount;
if(remainingAmount>=minTrimLength){
if(rcomp){
TrimRead.trimByAmount(r, trimAmount, 0, 1, false);
}else{
TrimRead.trimByAmount(r, 0, trimAmount, 1, false);
}
}else if(trimAmount>=minTrimLength){
if(rcomp){
TrimRead.trimByAmount(r, 0, remainingAmount, 1, false);
}else{
TrimRead.trimByAmount(r, remainingAmount, 0, 1, false);
}
}else{
//do nothing
}
}
return true;
}
public static byte[] softClipMatch(byte[] match, int minClipLength, boolean allowMutation){
final int matchScore=100;
final int subScore=-200;
final int subScore2=-100;
final int insScore=-200;
final int delScore=-200;
final int delScore2=-10;
final int clipScore=-1;
final int nScore=1;
int insCount=0;
int delCount=0;
long score=0;
long maxScore=0;
int maxPos=-1;
int maxStart=-1;
int currentStart=-1;
byte current='?';
for(int mpos=0; mpos<match.length; mpos++){
final byte m=match[mpos];
// long prevScore=score;
if(m=='m' || m=='N' || m=='R'){
if(score==0){currentStart=mpos;}
score=score+(m=='m' ? matchScore : nScore);
if(score>maxScore){
maxScore=score;
maxPos=mpos;
maxStart=currentStart;
}
}else{
if(m=='S' || m=='s'){
score=score+(m==current ? subScore2 : subScore);
}else if(m=='D'){
score=score+(m==current ? delScore2 : delScore);
delCount++;
}else if(m=='I' || m=='X' || m=='Y'){
score=score+insScore;
insCount++;
}else if(m=='C'){
score=score+clipScore;
}
score=Tools.max(0, score);
}
current=m;
}
if(maxScore<1){return match;}
final int leftClipM=maxStart;
final int rightClipM=(match.length-maxPos-1);
int leftClip=0, rightClip=0;
for(int i=0; i<match.length; i++){
byte m=match[i];
if(i<maxStart){
leftClip+=(m=='D' ? 0 : 1);
}else if(i>maxPos){
rightClip+=(m=='D' ? 0 : 1);
}
}
if(leftClip<minClipLength && rightClip<minClipLength){return match;}
if(delCount==0){
final byte[] array=allowMutation ? match : match.clone();
for(int i=0; i<leftClip; i++){array[i]='C';}
for(int i=0, j=array.length-1; i<rightClip; i++, j--){array[j]='C';}
return array;
}
ByteBuilder bb=new ByteBuilder(match.length);
if(leftClip>=minClipLength){
for(int mpos=0, processed=0; mpos<match.length; mpos++){
byte m=match[mpos];
if(m=='D'){
if(mpos>=leftClipM){bb.append(m);}
}else{
bb.append(mpos<leftClipM ? (byte)'C' : m);
processed++;
}
}
}else{
bb.append(match);
}
if(rightClip>=minClipLength){
bb.reverseInPlace();
byte[] temp=bb.toBytes();
bb.clear();
for(int mpos=0, processed=0; mpos<temp.length; mpos++){
byte m=temp[mpos];
if(m=='D'){
if(mpos>=rightClipM){bb.append(m);}
}else{
bb.append(mpos<rightClipM ? (byte)'C' : m);
processed++;
}
}
bb.reverseInPlace();
}
// System.out.println(new String(match)+"\n"+bb.toString()+"\n");
// System.out.println("length="+match.length);
// System.out.println("maxScore="+maxScore);
// System.out.println("maxPos="+maxPos);
// System.out.println("maxStart="+maxStart);
// System.out.println("leftClip="+leftClip);
// System.out.println("rightClip="+rightClip);
// System.out.println("leftClipM="+leftClipM);
// System.out.println("rightClipM="+rightClipM);
// System.out.println();
return bb.toBytes();
}
/*--------------------------------------------------------------*/
private String in1=null;
private String out1=null;
private String extin=null;
private String extout=null;
private String junctionFile="junctions_k%.txt";
/*--------------------------------------------------------------*/
private long maxReads=-1;
private int minClipLength=8;
private int minTrimLength=25;
private int minCount=2;
private float minFraction=0.0005f;
boolean printKmers=true;
boolean trim=true;
private long[][] counts;
private long[][] leftCounts;
private long[][] rightCounts;
/*--------------------------------------------------------------*/
private final FileFormat ffin1;
private final FileFormat ffout1;
/*--------------------------------------------------------------*/
private PrintStream outstream=System.err;
public static boolean verbose=false;
public boolean errorState=false;
private boolean overwrite=true;
private boolean append=false;
}
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