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package pacbio;
import java.util.ArrayList;
import aligner.MultiStateAligner9PacBioAdapter;
import aligner.MultiStateAligner9PacBioAdapter2;
import dna.AminoAcid;
import dna.Data;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import shared.KillSwitch;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FastaReadInputStream;
import stream.Read;
import structures.ListNum;
import tracker.ReadStats;
/**
* Increased sensitivity to nearby adapters.
* @author Brian Bushnell
* @date Nov 5, 2012
*
*/
public class RemoveAdapters2 {
public static void main(String[] args){
{//Preparse block for help, config files, and outstream
PreParser pp=new PreParser(args, new Object() { }.getClass().getEnclosingClass(), false);
args=pp.args;
//outstream=pp.outstream;
}
boolean verbose=false;
String in1=null;
String in2=null;
long maxReads=-1;
String outname1=null;
String outname2=null;
String query=pacbioAdapter;
Shared.capBufferLen(20);
boolean splitReads=true;
for(int i=0; i<args.length; i++){
final String arg=args[i];
final String[] split=arg.split("=");
String a=split[0].toLowerCase();
String b=split.length>1 ? split[1] : null;
if(Parser.parseCommonStatic(arg, a, b)){
//do nothing
}else if(Parser.parseZip(arg, a, b)){
//do nothing
}else if(Parser.parseQuality(arg, a, b)){
//do nothing
}else if(Parser.parseFasta(arg, a, b)){
//do nothing
}else if(a.equals("path") || a.equals("root") || a.equals("tempdir")){
Data.setPath(b);
}else if(a.equals("usealtmsa")){
USE_ALT_MSA=Parse.parseBoolean(b);
}else if(a.equals("fasta") || a.equals("in") || a.equals("input") || a.equals("in1") || a.equals("input1")){
in1=b;
if(b.indexOf('#')>-1){
in1=b.replace("#", "1");
in2=b.replace("#", "2");
}
}else if(a.equals("in2") || a.equals("input2")){
in2=b;
}else if(a.equals("query") || a.equals("adapter")){
query=b;
}else if(a.equals("split")){
splitReads=Parse.parseBoolean(b);
}else if(a.equals("plusonly")){
boolean x=Parse.parseBoolean(b);
if(x){TRY_PLUS=true; TRY_MINUS=false;}
}else if(a.equals("minusonly")){
boolean x=Parse.parseBoolean(b);
if(x){TRY_PLUS=false; TRY_MINUS=true;}
}else if(a.startsWith("mincontig")){
minContig=Integer.parseInt(b);
}else if(a.equals("append") || a.equals("app")){
append=ReadStats.append=Parse.parseBoolean(b);
}else if(a.equals("overwrite") || a.equals("ow")){
overwrite=Parse.parseBoolean(b);
System.out.println("Set overwrite to "+overwrite);
}else if(a.equals("threads") || a.equals("t")){
if(b.equalsIgnoreCase("auto")){THREADS=Shared.LOGICAL_PROCESSORS;}
else{THREADS=Integer.parseInt(b);}
System.out.println("Set threads to "+THREADS);
}else if(a.equals("reads") || a.equals("maxreads")){
maxReads=Parse.parseKMG(b);
}else if(a.startsWith("outname") || a.startsWith("outfile") || a.equals("out")){
if(b==null || b.equalsIgnoreCase("null") || b.equalsIgnoreCase("none") || split.length==1){
System.out.println("No output file.");
outname1=null;
OUTPUT_READS=false;
}else{
OUTPUT_READS=true;
if(b.indexOf('#')>-1){
outname1=b.replace('#', '1');
outname2=b.replace('#', '2');
}else{
outname1=b;
}
}
}else if(a.equals("minratio")){
MINIMUM_ALIGNMENT_SCORE_RATIO=Float.parseFloat(b);
System.out.println("Set MINIMUM_ALIGNMENT_SCORE_RATIO to "+MINIMUM_ALIGNMENT_SCORE_RATIO);
}else if(a.equals("suspectratio")){
SUSPECT_RATIO=Float.parseFloat(b);
}else if(a.startsWith("verbose")){
verbose=Parse.parseBoolean(b);
}else{
throw new RuntimeException("Unknown parameter: "+args[i]);
}
}
{//Process parser fields
Parser.processQuality();
}
assert(FastaReadInputStream.settingsOK());
if(in1==null){throw new RuntimeException("Please specify input file.");}
final ConcurrentReadInputStream cris;
{
FileFormat ff1=FileFormat.testInput(in1, FileFormat.FASTQ, null, true, true);
FileFormat ff2=FileFormat.testInput(in2, FileFormat.FASTQ, null, true, true);
cris=ConcurrentReadInputStream.getReadInputStream(maxReads, true, ff1, ff2);
// if(verbose){System.err.println("Started cris");}
// cris.start(); //4567
// th.start();
}
boolean paired=cris.paired();
if(verbose){System.err.println("Paired: "+paired);}
ConcurrentReadOutputStream ros=null;
if(OUTPUT_READS){
final int buff=(!ordered ? THREADS : Tools.max(24, 2*THREADS));
FileFormat ff1=FileFormat.testOutput(outname1, FileFormat.FASTQ, null, true, overwrite, append, ordered);
FileFormat ff2=FileFormat.testOutput(outname2, FileFormat.FASTQ, null, true, overwrite, append, ordered);
ros=ConcurrentReadOutputStream.getStream(ff1, ff2, buff, null, true);
}
process(cris, ros, query, splitReads);
}
public static void process(ConcurrentReadInputStream cris, ConcurrentReadOutputStream ros, String query, boolean split){
Timer t=new Timer();
cris.start(); //4567
System.out.println("Started read stream.");
if(ros!=null){
ros.start();
System.out.println("Started output threads.");
}
ProcessThread[] pts=new ProcessThread[THREADS];
for(int i=0; i<pts.length; i++){
pts[i]=new ProcessThread(cris, ros, MINIMUM_ALIGNMENT_SCORE_RATIO, query, split);
pts[i].start();
}
System.out.println("Started "+pts.length+" processing thread"+(pts.length==1 ? "" : "s")+".");
for(int i=0; i<pts.length; i++){
ProcessThread pt=pts[i];
synchronized(pt){
while(pt.getState()!=Thread.State.TERMINATED){
try {
pt.join();
} catch (InterruptedException e) {
// TODO Auto-generated catch block
e.printStackTrace();
}
}
}
if(i==0){
System.out.print("Detecting finished threads: 0");
}else{
System.out.print(", "+i);
}
}
System.out.println('\n');
ReadWrite.closeStreams(cris, ros);
printStatistics(pts);
t.stop();
System.out.println("Time: \t"+t);
}
public static void printStatistics(ProcessThread[] pts){
long plusAdaptersFound=0;
long minusAdaptersFound=0;
long goodReadsFound=0;
long badReadsFound=0;
long truepositive=0;
long truenegative=0;
long falsepositive=0;
long falsenegative=0;
long expected=0;
long unexpected=0;
long basesIn=0;
long basesOut=0;
long readsOut=0;
for(ProcessThread pt : pts){
plusAdaptersFound+=pt.plusAdaptersFound;
minusAdaptersFound+=pt.minusAdaptersFound;
goodReadsFound+=pt.goodReadsFound;
badReadsFound+=pt.badReadsFound;
basesIn+=pt.basesIn;
basesOut+=pt.basesOut;
readsOut+=pt.readsOut;
truepositive+=pt.truepositive;
truenegative+=pt.truenegative;
falsepositive+=pt.falsepositive;
falsenegative+=pt.falsenegative;
expected+=pt.expected;
unexpected+=pt.unexpected;
}
long totalReads=goodReadsFound+badReadsFound;
long totalAdapters=plusAdaptersFound+minusAdaptersFound;
if(expected<1){expected=1;}
if(unexpected<1){unexpected=1;}
System.out.println("Reads In: \t"+totalReads+" \t("+basesIn+" bases, avg length "+(basesIn/totalReads)+")");
System.out.println("Good reads: \t"+goodReadsFound);
System.out.println("Bad reads: \t"+badReadsFound+" \t("+totalAdapters+" adapters)");
System.out.println("Plus adapters: \t"+plusAdaptersFound);
System.out.println("Minus adapters: \t"+minusAdaptersFound);
System.out.println("Adapters per megabase: \t"+Tools.format("%.3f",totalAdapters*1000000f/basesIn));
if(readsOut>0){System.out.println("Reads Out: \t"+readsOut+" \t("+basesOut+" bases, avg length "+(basesOut/readsOut)+")");}
System.out.println();
if(truepositive>0 || truenegative>0 || falsepositive>0 || falsenegative>0){
System.out.println("Adapters Expected: \t"+expected);
System.out.println("True Positive: \t"+truepositive+" \t"+Tools.format("%.3f%%", truepositive*100f/expected));
System.out.println("True Negative: \t"+truenegative+" \t"+Tools.format("%.3f%%", truenegative*100f/unexpected));
System.out.println("False Positive: \t"+falsepositive+" \t"+Tools.format("%.3f%%", falsepositive*100f/unexpected));
System.out.println("False Negative: \t"+falsenegative+" \t"+Tools.format("%.3f%%", falsenegative*100f/expected));
}
}
private static class ProcessThread extends Thread{
public ProcessThread(ConcurrentReadInputStream cris_,
ConcurrentReadOutputStream ros_, float minRatio_, String query_, boolean split_) {
cris=cris_;
ros=ros_;
minRatio=minRatio_;
query1=query_.getBytes();
query2=AminoAcid.reverseComplementBases(query1);
ALIGN_ROWS=query1.length+1;
ALIGN_COLUMNS=ALIGN_ROWS*3+20;
SPLIT=split_;
stride=(int)(query1.length*0.95f);
window=(int)(query1.length*2.5f+10);
assert(window<ALIGN_COLUMNS);
msa=new MultiStateAligner9PacBioAdapter(ALIGN_ROWS, ALIGN_COLUMNS);
msa2=USE_ALT_MSA ? new MultiStateAligner9PacBioAdapter2() : null;
maxSwScore=msa.maxQuality(query1.length);
minSwScore=(int)(maxSwScore*MINIMUM_ALIGNMENT_SCORE_RATIO);
minSwScoreSuspect=(int)(maxSwScore*Tools.min(MINIMUM_ALIGNMENT_SCORE_RATIO*SUSPECT_RATIO, MINIMUM_ALIGNMENT_SCORE_RATIO-((1-SUSPECT_RATIO)*.2f)));
maxImperfectSwScore=msa.maxImperfectScore(query1.length);
suspectMidpoint=(minSwScoreSuspect+minSwScore)/2;
}
@Override
public void run(){
ListNum<Read> ln=cris.nextList();
ArrayList<Read> readlist=ln.list;
while(!readlist.isEmpty()){
//System.err.println("Got a list of size "+readlist.size());
for(int i=0; i<readlist.size(); i++){
Read r=readlist.get(i);
if(r.length()<minContig && (r.mate==null || r.mateLength()<minContig)){
readlist.set(i, null);
}else{
//System.out.println("Got read: "+r.toText());
//System.out.println("Synthetic: "+r.synthetic());
if(r.synthetic()){syntheticReads++;}
processRead(r);
if(r.mate!=null){processRead(r.mate);}
}
}
// System.err.println("outputStream = "+outputStream==null ? "null" : "real");
if(ros!=null){ //Important to send all lists to output, even empty ones, to keep list IDs straight.
if(DONT_OUTPUT_BROKEN_READS){removeDiscarded(readlist);}
for(Read r : readlist){
if(r!=null){
r.obj=null;//Not sure what r.obj is here
assert(r.bases!=null);
if(r.sites!=null && r.sites.isEmpty()){r.sites=null;}
}
}
// System.err.println("Adding list of length "+readlist.size());
ArrayList<Read> out=SPLIT ? split(readlist) : readlist;
for(Read r : out){
if(r!=null){
Read r2=r.mate;
basesOut+=r.length();
readsOut++;
if(r2!=null){
basesOut+=r2.length();
readsOut++;
}
}
}
ros.add(out, ln.id);
}
cris.returnList(ln.id, readlist.isEmpty());
//System.err.println("Waiting on a list...");
ln=cris.nextList();
readlist=ln.list;
}
//System.err.println("Returning a list... (final)");
assert(readlist.isEmpty());
cris.returnList(ln.id, readlist.isEmpty());
}
/**
* @param readlist
* @return
*/
private static ArrayList<Read> split(ArrayList<Read> in) {
ArrayList<Read> out=new ArrayList<Read>(in.size());
for(Read r : in){
if(r!=null){
// assert(r.mate==null);
if(!r.hasAdapter()){out.add(r);}
else{out.addAll(split(r));}
Read r2=r.mate;
if(r2!=null){
if(!r2.hasAdapter()){out.add(r2);}
else{out.addAll(split(r2));}
}
}
}
return out;
}
/**
* @param r
* @return
*/
private static ArrayList<Read> split(Read r) {
ArrayList<Read> sections=new ArrayList<Read>();
int lastX=-1;
for(int i=0; i<r.length(); i++){
if(r.bases[i]=='X'){
if(i-lastX>minContig){
byte[] b=KillSwitch.copyOfRange(r.bases, lastX+1, i);
byte[] q=r.quality==null ? null : KillSwitch.copyOfRange(r.quality, lastX+1, i);
Read r2=new Read(b, q, r.id+"_"+(lastX+1), r.numericID);
sections.add(r2);
}
lastX=i;
}
}
int i=r.length();
if(i-lastX>minContig){
byte[] b=KillSwitch.copyOfRange(r.bases, lastX+1, i);
byte[] q=r.quality==null ? null : KillSwitch.copyOfRange(r.quality, lastX+1, i);
Read r2=new Read(b, q, r.id+"_"+(lastX+1), r.numericID);
sections.add(r2);
}
return sections;
}
/**
* @param r
*/
private int processRead(Read r) {
int begin=0;
while(begin<r.length() && r.bases[begin]=='N'){begin++;} //Skip reads made of 'N'
if(begin>=r.length()){return 0;}
basesIn+=r.length();
final byte[] array=npad(r.bases, npad);
int lim=array.length-npad-stride;
int plusFound=0;
int minusFound=0;
int lastSuspect=-1;
int lastConfirmed=-1;
for(int i=begin; i<lim; i+=stride){
int j=Tools.min(i+window, array.length-1);
if(j-i<window){
lim=0; //Last loop cycle
// i=Tools.max(0, array.length-2*query1.length);
}
if(TRY_MINUS){
int[] rvec=msa.fillAndScoreLimited(query2, array, i, j, minSwScoreSuspect);
if(rvec!=null && rvec[0]>=minSwScoreSuspect){
int score=rvec[0];
int start=rvec[1];
int stop=rvec[2];
assert(score>=minSwScoreSuspect);
if((i==0 || start>i) && (j==array.length-1 || stop<j)){
boolean kill=(score>=minSwScore ||
(score>=suspectMidpoint && lastSuspect>0 && start>=lastSuspect && start-lastSuspect<suspectDistance) ||
(lastConfirmed>0 && start>=lastConfirmed && start-lastConfirmed<suspectDistance));
if(!kill && USE_LOCALITY && array.length-stop>window){//Look ahead
rvec=msa.fillAndScoreLimited(query2, array, stop, stop+window, minSwScoreSuspect);
if(rvec!=null){
if(score>=suspectMidpoint && rvec[0]>=minSwScoreSuspect && rvec[1]-stop<suspectDistance){kill=true;}
else if(score>=minSwScoreSuspect && rvec[0]>=minSwScore && rvec[1]-stop<suspectDistance){kill=true;}
}
}
if(!kill && USE_ALT_MSA){//Try alternate msa
rvec=msa2.fillAndScoreLimited(query2, array, Tools.max(0, start-4), Tools.min(stop+4, array.length-1), minSwScoreSuspect);
if(rvec!=null && rvec[0]>=(minSwScore)){kill=true;}
}
if(kill){
// System.out.println("-:"+score+", "+minSwScore+", "+minSwScoreSuspect+"\t"+lastSuspect+", "+start+", "+stop);
minusFound++;
for(int x=Tools.max(0, start); x<=stop; x++){array[x]='X';}
if(USE_LOCALITY && score>=minSwScore){lastConfirmed=Tools.max(lastConfirmed, stop);}
}
}
// System.out.println("Set lastSuspect="+stop+" on score "+score);
if(USE_LOCALITY){lastSuspect=Tools.max(lastSuspect, stop);}
}
}
if(TRY_PLUS){
int[] rvec=msa.fillAndScoreLimited(query1, array, i, j, minSwScoreSuspect);
if(rvec!=null && rvec[0]>=minSwScoreSuspect){
int score=rvec[0];
int start=rvec[1];
int stop=rvec[2];
if((i==0 || start>i) && (j==array.length-1 || stop<j)){
boolean kill=(score>=minSwScore ||
(score>=suspectMidpoint && lastSuspect>0 && start>=lastSuspect && start-lastSuspect<suspectDistance) ||
(lastConfirmed>0 && start>=lastConfirmed && start-lastConfirmed<suspectDistance));
if(!kill && USE_LOCALITY && array.length-stop>window){//Look ahead
rvec=msa.fillAndScoreLimited(query1, array, stop, stop+window, minSwScoreSuspect);
if(rvec!=null){
if(score>=suspectMidpoint && rvec[0]>=minSwScoreSuspect && rvec[1]-stop<suspectDistance){kill=true;}
else if(score>=minSwScoreSuspect && rvec[0]>=minSwScore && rvec[1]-stop<suspectDistance){kill=true;}
}
}
if(!kill && USE_ALT_MSA){//Try alternate msa
rvec=msa2.fillAndScoreLimited(query1, array, Tools.max(0, start-4), Tools.min(stop+4, array.length-1), minSwScoreSuspect);
if(rvec!=null && rvec[0]>=(minSwScore)){kill=true;}
}
if(kill){
// System.out.println("+:"+score+", "+minSwScore+", "+minSwScoreSuspect+"\t"+lastSuspect+", "+start+", "+stop);
plusFound++;
for(int x=Tools.max(0, start); x<=stop; x++){array[x]='X';}
if(USE_LOCALITY && score>=minSwScore){lastConfirmed=Tools.max(lastConfirmed, stop);}
}
}
// System.out.println("Set lastSuspect="+stop+" on score "+score);
if(USE_LOCALITY){lastSuspect=Tools.max(lastSuspect, stop);}
}
}
}
int found=plusFound+minusFound;
// if(r.synthetic()){
// if(/*r.hasadapter() && */(r.numericID&3)==0){
// if(plusFound>0){truepositive++;}else{falsenegative++;}
// if(plusFound>1){falsepositive+=(plusFound-1);}
// falsepositive+=minusFound;
// expected++;
// }else if(/*r.hasadapter() && */(r.numericID&3)==1){
// if(minusFound>0){truepositive++;}else{falsenegative++;}
// if(minusFound>1){falsepositive+=(minusFound-1);}
// falsepositive+=plusFound;
// expected++;
// }else{
// falsepositive=falsepositive+plusFound+minusFound;
// if(plusFound+minusFound==0){truenegative++;}
// unexpected++;
// }
// }
if(r.synthetic()){
if(/*r.hasadapter() && */(r.numericID&3)==0){
if(found>0){truepositive++;}else{falsenegative++;}
if(found>1){falsepositive+=(found-1);}
expected++;
}else if(/*r.hasadapter() && */(r.numericID&3)==1){
if(found>0){truepositive++;}else{falsenegative++;}
if(found>1){falsepositive+=(found-1);}
expected++;
}else{
falsepositive+=found;
if(found==0){truenegative++;}
unexpected++;
}
}
plusAdaptersFound+=plusFound;
minusAdaptersFound+=minusFound;
if(found>0){
for(int i=npad, j=0; j<r.length(); i++, j++){r.bases[j]=array[i];}
if(DONT_OUTPUT_BROKEN_READS){r.setDiscarded(true);}
badReadsFound++;
}else{
goodReadsFound++;
}
r.setHasAdapter(found>0);
return found;
}
private byte[] npad(final byte[] array, final int pad){
final int len=array.length+2*pad;
if(padbuffer==null || padbuffer.length!=len){padbuffer=new byte[len];}
byte[] r=padbuffer;
for(int i=0; i<pad; i++){r[i]='N';}
for(int i=pad, j=0; j<array.length; i++, j++){r[i]=array[j];}
for(int i=array.length+pad, limit=Tools.min(r.length, len+50); i<limit; i++){r[i]='N';}
padbuffer=null; //Kills the buffer. Causes more memory allocation, but better cache/NUMA locality if threads move around.
return r;
}
private byte[] padbuffer=null;
private final byte[] query1, query2;
private final ConcurrentReadInputStream cris;
private final ConcurrentReadOutputStream ros;
private final float minRatio;
private final MultiStateAligner9PacBioAdapter msa;
private final MultiStateAligner9PacBioAdapter2 msa2;
private final int ALIGN_ROWS;
private final int ALIGN_COLUMNS;
private final int stride;
private final int window;
private final boolean SPLIT;
long plusAdaptersFound=0;
long minusAdaptersFound=0;
long goodReadsFound=0;
long badReadsFound=0;
long truepositive=0;
long truenegative=0;
long falsepositive=0;
long falsenegative=0;
long expected=0;
long unexpected=0;
long basesIn=0;
long basesOut=0;
long readsOut=0;
private final int maxSwScore;
private final int minSwScore;
private final int minSwScoreSuspect;
private final int suspectMidpoint;
private final int maxImperfectSwScore;
long syntheticReads=0;
}
private static int removeDiscarded(ArrayList<Read> list){
int removed=0;
for(int i=0; i<list.size(); i++){
Read r=list.get(i);
if(r.discarded()){
if(r.mate==null || r.mate.discarded()){
list.set(i, null);
removed++;
}
}
}
return removed;
}
public static boolean DONT_OUTPUT_BROKEN_READS;
/** Permission to overwrite existing files */
private static boolean overwrite=true;
/** Permission to append to existing files */
private static boolean append=false;
private static int THREADS=Shared.LOGICAL_PROCESSORS;
private static boolean OUTPUT_READS=false;
private static boolean ordered=false;
private static boolean PERFECTMODE=false;
private static float MINIMUM_ALIGNMENT_SCORE_RATIO=0.31f; //0.31f: At 250bp reads, approx 0.01% false-positive and 94% true-positive.
private static float SUSPECT_RATIO=0.85F;
public static boolean USE_LOCALITY=true;
public static boolean USE_ALT_MSA=true;
public static boolean TRY_PLUS=true;
public static boolean TRY_MINUS=true;
private static int npad=35;
public static int minContig=50;
public static int suspectDistance=100;
public static final String pacbioAdapter="ATCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGAT";
public static final String pacbioStandard_v1="TCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGAGAAGGCTGGGCAGGCTATGCACCCTGGTCCAGGTCAAA" +
"AGCTGCGGAACCCGCTAGCGGCCATCTTGGCCACTAGGGGTCCCGCAGATTCATATTGTCGTCTAGCATGCACAATGCTGCAAACCCAGCTTGCAATGCCCACAGCA" +
"AGCGGCCAATCTTTACGCCACGTTGAATTGTTTATTACCTGTGACTGGCTATGGCTTGCAACGCCACTCGTAAAACTAGTACTTTGCGGTTAGGGGAAGTAGACAAA" +
"CCCATTACTCCACTTCCCGGAAGTTCAACTCATTCCAACACGAAATAAAAGTAAACTCAACACCCCAAGCAGGCTATGTGGGGGGGTGATAGGGGTGGATTCTATTT" +
"CCTATCCCATCCCCTAGGATCTCAATTAAGTTACTAGCGAGTTAAATGTCTGTAGCGATCCCGTCAGTCCTATCGCGCGCATCAAGACCTGGTTGGTTGAGCGTGCA" +
"GTAGATCATCGATAAGCTGCGAGTTAGGTCATCCCAGACCGCATCTGGCGCCTAAACGTTCAGTGGTAGCTAAGGCGTCACCTTCGACTGTCTAAAGGCAATATGTC" +
"GTCCTTAGCTCCAAGTCCCTAGCAAGCGTGTCGGGTCTCTCTCTTTTCCTCCTCCTCCGTTGTTGTTGTTGAGAGAGACCCGACACGCTTGCTAGGGACTTGGAGCT" +
"AAGGACGACATATTGCCTTTAGACAGTCGAAGGTGACGCCTTAGCTACCACTGAACGTTTAGGCGCCAGATGCGGTCTGGGATGACCTAACTCGCAGCTTATCGATG" +
"ATCTACTGCACGCTCAACCAACCAGGTCTTGATGCGCGCGATAGGACTGACGGGATCGCTACAGACATTTAACTCGCTAGTAACTTAATTGAGATCCTAGGGGATGG" +
"GATAGGAAATAGAATCCACCCCTATCACCCCCCCACATAGCCTGCTTGGGGTGTTGAGTTTACTTTTATTTCGTGTTGGAATGAGTTGAACTTCCGGGAAGTGGAGT" +
"AATGGGTTTGTCTACTTCCCCTAACCGCAAAGTACTAGTTTTACGAGTGGCGTTGCAAGCCATAGCCAGTCACAGGTAATAAACAATTCAACGTGGCGTAAAGATTG" +
"GCCGCTTGCTGTGGGCATTGCAAGCTGGGTTTGCAGCATTGTGCATGCTAGACGACAATATGAATCTGCGGGACCCCTAGTGGCCAAGATGGCCGCTAGCGGGTTCC" +
"GCAGCTTTTGACCTGGACCAGGGTGCATAGCCTGCCCAGCCTTCTCTCTCTCTTT";
}
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