Mercurial > repos > rliterman > csp2
view CSP2/CSP2_env/env-d9b9114564458d9d-741b3de822f2aaca6c6caa4325c4afce/opt/bbmap-39.01-1/current/prok/GeneCaller.java @ 68:5028fdace37b
planemo upload commit 2e9511a184a1ca667c7be0c6321a36dc4e3d116d
author | jpayne |
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date | Tue, 18 Mar 2025 16:23:26 -0400 |
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package prok; import java.io.PrintStream; import java.util.ArrayList; import java.util.Arrays; import java.util.Collections; import aligner.SingleStateAlignerFlat2; import aligner.SingleStateAlignerFlat3; import aligner.SingleStateAlignerFlatFloat; import dna.AminoAcid; import shared.KillSwitch; import shared.Tools; import stream.Read; import structures.FloatList; import structures.IntList; import structures.LongHashSet; /** * This class calls genes within a single thread. * @author Brian Bushnell * @date Sep 24, 2018 * */ public class GeneCaller extends ProkObject { /*--------------------------------------------------------------*/ /*---------------- Init ----------------*/ /*--------------------------------------------------------------*/ GeneCaller(int minLen_, int maxOverlapSameStrand_, int maxOverlapOppositeStrand_, float minStartScore_, float minStopScore_, float minInnerScore_, float minOrfScore_, float minAvgScore_, GeneModel pgm_){ minLen=minLen_; maxOverlapSameStrand=maxOverlapSameStrand_; maxOverlapOppositeStrand=maxOverlapOppositeStrand_; pgm=pgm_; minStartScore=minStartScore_; minStopScore=minStopScore_; minInnerScore=minInnerScore_; minOrfScore=minOrfScore_; minAvgScore=minAvgScore_; } /*--------------------------------------------------------------*/ /*---------------- Outer Methods ----------------*/ /*--------------------------------------------------------------*/ public ArrayList<Orf> callGenes(Read r){ return callGenes(r, pgm); } public ArrayList<Orf> callGenes(Read r, GeneModel pgm_){ pgm=pgm_; final String name=r.id; final byte[] bases=r.bases; //Lists of all longest orfs per frame ArrayList<Orf>[] frameLists=makeOrfs(name, bases, minLen); //Lists of all high-scoring orfs per frame, with potentially multiple orfs sharing stops. ArrayList<Orf>[] brokenLists=breakOrfs(frameLists, bases); ArrayList<Orf>[] rnaLists=null; final int rlen=r.length(); if(calltRNA || (call16S && rlen>800) || (call23S && rlen>1500) || call5S || (call18S && rlen>1000)){ rnaLists=makeRnas(name, bases); brokenLists[0].addAll(rnaLists[0]); brokenLists[3].addAll(rnaLists[1]); Collections.sort(brokenLists[0]); Collections.sort(brokenLists[3]); } boolean printAllOrfs=false; boolean printRnas=false; if(printAllOrfs){ ArrayList<Orf> temp=new ArrayList<Orf>(); for(ArrayList<Orf> broken : brokenLists){ temp.addAll(broken); } Collections.sort(temp); return temp; } if(printRnas && rnaLists!=null){ ArrayList<Orf> temp=new ArrayList<Orf>(); for(ArrayList<Orf> list : rnaLists){ temp.addAll(list); } Collections.sort(temp); return temp; } stCds2.add(brokenLists); //Find the optimal path through Orfs ArrayList<Orf> path=findPath(brokenLists, bases); // geneStartsOut+=path.size(); if(callCDS){stCdsPass.add(path);} if(calltRNA){sttRNA.add(path);} if(call16S){st16s.add(path);} if(call23S){st23s.add(path);} if(call5S){st5s.add(path);} if(call18S){st18s.add(path);} return path; } /*--------------------------------------------------------------*/ /*---------------- Inner Methods ----------------*/ /*--------------------------------------------------------------*/ /** * Generates lists of all max-length non-overlapping Orfs per frame. * There IS overlap between frames. * All Orfs come out flipped to + orientation. * */ ArrayList<Orf>[] makeRnas(String name, byte[] bases){ @SuppressWarnings("unchecked") ArrayList<Orf>[] array=new ArrayList[2]; array[0]=new ArrayList<Orf>(); array[1]=new ArrayList<Orf>(); final float[] scores=new float[bases.length]; final int[] kmersSeen=(lsuKmers==null && ssuKmers==null && trnaKmers==null && r5SKmers==null) ? null : new int[bases.length]; for(int strand=0; strand<2; strand++){ for(StatsContainer sc : pgm.rnaContainers){ if(ProkObject.callType(sc.type)){ ArrayList<Orf> list=makeRnasForStrand(name, bases, strand, sc, scores, (sc.kmerSet()==null ? null : kmersSeen), false, -1);//TODO: Make this loop through all RNA types if(strand==1 && list!=null){ for(Orf orf : list){ assert(orf.strand==strand); orf.flip(); } } if(list!=null){array[strand].addAll(list);} } } Collections.sort(array[strand]); AminoAcid.reverseComplementBasesInPlace(bases); } return array; } /** Designed for quickly calling a single SSU */ public Orf makeRna(String name, byte[] bases, int type){ final float[] scores=new float[bases.length];//TODO: Big and slow; make a FloatList? StatsContainer sc=pgm.allContainers[type]; final int[] kmersSeen=(sc.kmerSet()==null ? null : new int[bases.length]);//TODO: IntList? int strand=0; ArrayList<Orf> list=makeRnasForStrand(name, bases, strand, sc, scores, kmersSeen, true, -1); final Orf best1=pickBest(list); assert(best1==null || best1.start>=0 && best1.stop<bases.length) : bases.length+"\n"+best1; if(best1!=null && best1.orfScore>-999){return best1;} strand++; AminoAcid.reverseComplementBasesInPlace(bases); list=makeRnasForStrand(name, bases, strand, sc, scores, kmersSeen, true, -1); AminoAcid.reverseComplementBasesInPlace(bases); if(strand==1 && list!=null){ for(Orf orf : list){ assert(orf.strand==strand); orf.flip(); } } final Orf best2=pickBest(list); assert(best2==null || best2.start>=0 && best2.stop<bases.length) : bases.length+"\n"+best2; if(best2!=null && best2.orfScore>-999){return best2;} return best1!=null ? best1 : best2; } final Orf pickBest(ArrayList<Orf> list){ if(list==null){return null;} Orf best=null; for(Orf orf : list){ if(best==null || orf.orfScore>best.orfScore){ best=orf; } } return best; } /** * Generates lists of all max-length non-overlapping Orfs per frame. * There IS overlap between frames. * All Orfs come out flipped to + orientation. * */ static ArrayList<Orf>[] makeOrfs(String name, byte[] bases, int minlen){ @SuppressWarnings("unchecked") ArrayList<Orf>[] array=new ArrayList[6]; for(int strand=0; strand<2; strand++){ for(int frame=0; frame<3; frame++){ ArrayList<Orf> list=makeOrfsForFrame(name, bases, frame, strand, minlen); array[frame+3*strand]=list; if(strand==1 && list!=null){ for(Orf orf : list){ assert(orf.frame==frame); assert(orf.strand==strand); orf.flip(); } } } AminoAcid.reverseComplementBasesInPlace(bases); } return array; } /** * Dynamic programming phase. * @param frameLists * @param bases * @return */ private ArrayList<Orf> findPath(ArrayList<Orf>[] frameLists, byte[] bases){ ArrayList<Orf> all=new ArrayList<Orf>(); for(ArrayList<Orf> list : frameLists){all.addAll(list);} if(all.isEmpty()){return all;} Collections.sort(all); for(Orf orf : all){ orf.pathScorePlus=-999999; orf.pathScoreMinus=-999999; } int[] lastPositionScored=KillSwitch.allocInt1D(6); Arrays.fill(lastPositionScored, -1); //Index of highest-scoring ORF in this frame, with prev on the plus strand int[] bestIndexPlus=KillSwitch.allocInt1D(6); //Index of highest-scoring ORF in this frame, with prev on the minus strand int[] bestIndexMinus=KillSwitch.allocInt1D(6); //Highest-scoring ORF in this frame, with prev on the plus strand Orf[] bestOrfPlus=new Orf[6]; //Highest-scoring ORF in this frame, with prev on the minus strand Orf[] bestOrfMinus=new Orf[6]; int[][] bestIndex=new int[][] {bestIndexPlus, bestIndexMinus}; Orf[][] bestOrf=new Orf[][] {bestOrfPlus, bestOrfMinus}; for(Orf orf : all){ final int myListNum=3*orf.strand+orf.frame; calcPathScore(orf, frameLists, lastPositionScored, bestIndex); if(bestOrfPlus[myListNum]==null || orf.pathScorePlus>=bestOrfPlus[myListNum].pathScorePlus){ bestOrfPlus[myListNum]=orf; bestIndexPlus[myListNum]=lastPositionScored[myListNum]; assert(frameLists[myListNum].get(lastPositionScored[myListNum])==orf); } if(bestOrfMinus[myListNum]==null || orf.pathScoreMinus>=bestOrfMinus[myListNum].pathScoreMinus){ bestOrfMinus[myListNum]=orf; bestIndexMinus[myListNum]=lastPositionScored[myListNum]; assert(frameLists[myListNum].get(lastPositionScored[myListNum])==orf); } } Orf best=bestOrf[0][0]; for(Orf[] array : bestOrf){ for(Orf orf : array){ if(best==null || (orf!=null && orf.pathScore()>best.pathScore())){ best=orf; } } } ArrayList<Orf> bestPath=new ArrayList<Orf>(); for(Orf orf=best; orf!=null; orf=orf.prev()){ bestPath.add(orf); if(orf.type==CDS){geneStartsOut++;} else if(orf.type==tRNA){tRNAOut++;} else if(orf.type==r16S){r16SOut++;} else if(orf.type==r23S){r23SOut++;} else if(orf.type==r5S){r5SOut++;} else if(orf.type==r18S){r18SOut++;} } Collections.sort(bestPath); return bestPath; } /** * Calculate the best path to this ORF. * @param orf * @param frameLists * @param lastPositionScored * @param bestIndex */ private void calcPathScore(Orf orf, ArrayList<Orf>[] frameLists, int[] lastPositionScored, int[][] bestIndex){ final int myListNum=3*orf.strand+orf.frame; // System.err.println("* "+orf); // System.err.println("* "+Arrays.toString(lastPositionScored)); // System.err.println(); for(int listStrand=0; listStrand<2; listStrand++){ for(int listFrame=0; listFrame<3; listFrame++){ int listNum=listFrame+3*listStrand; ArrayList<Orf> list=frameLists[listNum]; int lastPos=lastPositionScored[listNum]; int bestPos=bestIndex[listStrand][listNum]; if(listStrand==0){ calcPathScorePlus(orf, list, listStrand, lastPos, bestPos); }else{ calcPathScoreMinus(orf, list, listStrand, lastPos, bestPos); } } } // System.err.println(myListNum+", "+Arrays.toString(lastPositionScored)+", "+frameLists[myListNum].size()); lastPositionScored[myListNum]++; assert(frameLists[myListNum].get(lastPositionScored[myListNum])==orf) : myListNum+"\n"+orf+"\n"+frameLists[myListNum].get(lastPositionScored[myListNum])+"\n" +Arrays.toString(lastPositionScored)+"\n"+frameLists[myListNum].get(lastPositionScored[myListNum]+1); //These are sanity checks to make sure that the path did not break in the middle. //Safe to disable. // assert(orf.prevPlus!=null || orf.stop<100000); // assert(orf.prevMinus!=null || orf.stop<100000); // assert(orf.pathScore>-10) : orf.pathScore+"\n"+orf+"\n"+orf.prev+"\n"; } /** * Calculate the best path to this ORF from a plus-strand previous ORF. * @param orf * @param list * @param listStrand * @param lastPos * @param bestPos */ private void calcPathScorePlus(final Orf orf, final ArrayList<Orf> list, final int listStrand, final int lastPos, final int bestPos){ assert(listStrand==0); if(lastPos<0){ if(orf.prevPlus==null){ orf.pathScorePlus=orf.orfScore; orf.pathLengthPlus=1; } return; } if(list.isEmpty()){return;} // System.err.println("\nExamining \t"+orf+"\nlastPos="+lastPos+", bestPos="+bestPos+", sameFrame="+sameFrame); boolean found=false; final boolean sameStrand=(orf.strand==listStrand); final int maxOverlap=(sameStrand ? maxOverlapSameStrand : maxOverlapOppositeStrand); for(int i=lastPos, min=Tools.max(0, bestPos-lookbackPlus); i>=min || (i>0 && !found); i--){ Orf prev=list.get(i); assert(prev!=orf) : prev; // System.err.println("Comparing to \t"+prev); if(orf.isValidPrev(prev, maxOverlap)){ int overlap=Tools.max(0, prev.stop-orf.start+1); float orfScore=overlap==0 ? orf.orfScore : orf.calcOrfScore(overlap); final float prevScore=prev.pathScore(); final int prevLength=prev.pathLength(); float pathScore; final int pathLength; if(sameStrand){ pathLength=prevLength+1; pathScore=prevScore+orfScore; pathScore+=p0+p1*(Tools.mid(p5*(p2+pathLength), p6*(p3-pathLength), p4)); }else{ pathLength=1; pathScore=prev.pathScore()+orfScore; pathScore+=q1+Tools.mid(q2*prevLength, q3+q4*prevLength, q5); } if(overlap<1 && prevScore>0){found=true;} if(pathScore>=orf.pathScorePlus){ orf.pathScorePlus=pathScore; orf.prevPlus=prev; orf.pathLengthPlus=pathLength; // System.err.println("Set as best"); } } if(found && prev.stop<maxOverlap-2000 && orf.prevPlus!=null){ System.err.println("Breaking"); break; } } } /** * Calculate the best path to this ORF from a minus-strand previous ORF. * @param orf * @param list * @param listStrand * @param lastPos * @param bestPos */ private void calcPathScoreMinus(final Orf orf, final ArrayList<Orf> list, final int listStrand, final int lastPos, final int bestPos){ assert(listStrand==1); if(lastPos<0){ if(orf.prevMinus==null){ orf.pathScoreMinus=orf.orfScore; orf.pathLengthMinus=1; } return; } if(list.isEmpty()){return;} // System.err.println("\nExamining \t"+orf+"\nlastPos="+lastPos+", bestPos="+bestPos+", sameFrame="+sameFrame); boolean found=false; final boolean sameStrand=(orf.strand==listStrand); final int maxOverlap=(sameStrand ? maxOverlapSameStrand : maxOverlapOppositeStrand); for(int i=lastPos, min=Tools.max(0, bestPos-lookbackMinus); i>=min || (i>0 && !found); i--){ Orf prev=list.get(i); assert(prev!=orf) : prev; // System.err.println("Comparing to \t"+prev); if(orf.isValidPrev(prev, maxOverlap)){ int overlap=Tools.max(0, prev.stop-orf.start+1); float orfScore=overlap==0 ? orf.orfScore : orf.calcOrfScore(overlap); final float prevScore=prev.pathScore(); final int prevLength=prev.pathLength(); float pathScore; final int pathLength; if(sameStrand){ pathLength=prevLength+1; pathScore=prevScore+orfScore; pathScore+=p0+p1*(Tools.mid(p5*(p2+pathLength), p6*(p3-pathLength), p4)); }else{ pathLength=1; pathScore=prev.pathScore()+orfScore; pathScore+=q1+Tools.mid(q2*prevLength, q3+q4*prevLength, q5); } if(overlap<1 && prevScore>0){found=true;} if(pathScore>=orf.pathScoreMinus){ orf.pathScoreMinus=pathScore; orf.prevMinus=prev; orf.pathLengthMinus=pathLength; // System.err.println("Set as best"); } } if(found && prev.stop<maxOverlap-2000 && orf.prevMinus!=null){ System.err.println("Breaking"); break; } } } /** * Generates a list of maximal-length Orfs only (non-overlapping). * All Orfs come out in native orientation (unflipped). * */ static ArrayList<Orf> makeOrfsForFrame(String name, byte[] bases, int startFrame, int strand, int minlen){ // assert(false) : "TODO"; assert(minlen>=3); if(bases==null || bases.length<minlen){return null;} ArrayList<Orf> orfs=new ArrayList<Orf>(); if(!ProkObject.callCDS){return orfs;} // int mask=63; int code=0; int start=-2; int frame=0; int pos=startFrame; for(; pos<bases.length; pos++){ byte b=bases[pos]; int x=AminoAcid.baseToNumber[b]; // code=((code<<2)|x)&mask; code=((code<<2)|x); frame++; if(frame==3){ frame=0; if(start>=0){ if(GeneModel.isStopCodon(code) || code<0){//NOTE: This adds a stop codon wherever there are Ns. int len=pos-start+1; if(len>=minlen){ Orf f=new Orf(name, start, pos, strand, startFrame, bases, true, CDS); orfs.add(f); } start=-1; } }else{ if(start==-2 || (start<0 && GeneModel.isStartCodon(code))){ start=pos-2; } } code=0; } } //Add a stop codon at the sequence end. if(start>=0){ pos--; while(frame!=3 && frame!=-1){ pos--; frame--; } int len=pos-start+1; if(len>=minlen){ assert(pos<bases.length) : start+", "+pos+", "+bases.length; Orf f=new Orf(name, start, pos, strand, startFrame, bases, true, CDS); orfs.add(f); } } return orfs; } /** * Generates a list of maximal-length RNAs (non-overlapping). * All RNAs come out in native orientation (unflipped). * */ ArrayList<Orf> makeRnasForStrand(String name, byte[] bases, int strand, StatsContainer sc, float[] scores, int[] kmersSeen, boolean quitEarly, float bias){ final int window=sc.lengthAvg; if(bases==null || bases.length*2<window){return null;} ArrayList<Orf> orfs=new ArrayList<Orf>(sc.type==tRNA ? 32 : 8); final FrameStats inner=sc.inner; // final FrameStats start=sc.start; // final FrameStats stop=sc.stop; final int k=inner.k; final int mask=inner.mask; // final float invLen=sc.invLengthAvg; final int halfWindow=window/2; final int maxWindow=(int)(window*1.5f); final int maxWindow2=(int)(window*2.5f); // final int slop=Tools.max(50, window/8); int len=0; int kmer=0; float currentScore=0; float currentScoreAbs=0; bias=(bias>-1 ? bias : biases[sc.type]); final float maxBias=biases[sc.type]*1.45f; float thresh=cutoff1[sc.type]; float prevScore=0; int lastStart=0; float max=0; int maxPos=0; for(int pos=0; pos<bases.length; pos++){ final byte b=bases[pos]; assert(b>=0 && b<128) : "Invalid base b="+((int)b)+"; pos="+pos+"\n"+new String(bases)+"\n"; final int x=AminoAcid.baseToNumber[b]; kmer=((kmer<<2)|x)&mask; if(x>=0){ len++; if(len>=k){ float prob=inner.probs[0][kmer]; float dif=prob-bias;//Prob above 1 is more likely than average currentScoreAbs+=prob; currentScore=Tools.max(0, currentScore+dif); } if(currentScore>0){ if(currentScore>max){ max=currentScore; maxPos=pos; } if(prevScore<=0){ lastStart=pos; } }else{ int rnaLen=maxPos-lastStart; if(max>thresh && rnaLen>=halfWindow){ if(rnaLen>maxWindow){ if(bias<=maxBias){ orfs=null; float biasMult=(rnaLen>8*window ? 1.2f : rnaLen>4*window ? 1.1f : 1.05f); return makeRnasForStrand(name, bases, strand, sc, scores, kmersSeen, quitEarly, bias*biasMult); } } if(rnaLen<=maxWindow2){ Orf orf=new Orf(name, lastStart, maxPos, strand, 0, bases, false, sc.type); orfs.add(orf); orf.orfScore=max; if(verbose){ final int start2=(strand==0 ? lastStart : bases.length-maxPos-1); final int stop2=(strand==0 ? maxPos : bases.length-lastStart-1); System.err.println("Made Orf "+start2+"\t"+stop2+"\t"+max); } } } max=0; lastStart=pos; } // System.err.println("i="+i+"\tscore="+score+"\tmax="+max+"\tmaxPos="+maxPos+"\tprevScore="+prevScore+"\tlastStart="+lastStart); prevScore=currentScore; // if(pos>=223000 && pos<232000){ //// System.err.println("i="+i+"\tscore="+score+"\tmax="+max+"\tmaxPos="+maxPos+"\tprevScore="+prevScore+"\tlastStart="+lastStart); // System.out.println(pos+"\t"+currentScore); // } }else{ len=0; kmer=0; } scores[pos]=currentScoreAbs; } // System.err.println("size="+orfs.size()+", type="+Orf.typeStrings[sc.type]); { int rnaLen=maxPos-lastStart; if(max>thresh && rnaLen>=halfWindow){ if(rnaLen>maxWindow){ if(bias<=maxBias){ orfs=null; float biasMult=(rnaLen>8*window ? 1.2f : rnaLen>4*window ? 1.1f : 1.05f); return makeRnasForStrand(name, bases, strand, sc, scores, kmersSeen, quitEarly, bias*biasMult); } } if(rnaLen<=maxWindow2){ Orf orf=new Orf(name, lastStart, maxPos, strand, 0, bases, false, sc.type); orfs.add(orf); orf.orfScore=max; if(verbose){ final int start2=(strand==0 ? lastStart : bases.length-maxPos-1); final int stop2=(strand==0 ? maxPos : bases.length-lastStart-1); System.err.println(start2+"\t"+stop2+"\t"+max); } } } } if(kmersSeen!=null && orfs.size()>0 && sc.kmerSet()!=null){ fillKmersSeen(bases, kmersSeen, sc.kmerSet(), sc.kLongLen()); } float cutoff=cutoff2[sc.type]; for(int i=0; i<orfs.size(); i++){ Orf orf=orfs.get(i); // System.err.println(orf.orfScore); boolean good=refineRna(orf, bases, strand, sc, scores, kmersSeen); if(orf.orfScore<cutoff || !good){ if(verbose){System.err.println("REJECT: "+orf.toStringFlipped());} orfs.set(i, null); }else{ if(verbose){System.err.println("ACCEPT: "+orf.toStringFlipped());} if(quitEarly){ orfs.clear(); orfs.add(orf); return orfs; } } } Tools.condenseStrict(orfs); // assert(false); // for(int pos=0; pos<bases.length; pos++){ // final byte b=bases[pos]; // final int x=AminoAcid.baseToNumber[b]; // kmer=((kmer<<2)|x)&mask; // // if(x>=0){ // len++; // if(len>=k){ // float prob=inner.probs[0][kmer]; // float dif=prob-1.2f;//Prob above 1 is more likely than average // currentScore=Tools.max(0, currentScore+dif); // if(currentScore>0){ // currentStreak++; // }else{ // currentStreak=0; // } // if(currentScore>200 && currentStreak>1500 && currentStreak<1700){ // Orf orf=new Orf(name, pos-currentStreak-1, pos, strand, 0, bases, false); // orfs.add(orf); // orf.orfScore=currentScore; // orf.startScore=start.scorePoint(orf.start, bases); // orf.stopScore=stop.scorePoint(orf.stop, bases); // currentStreak=0; // currentScore=0; // } // } // }else{ // len=0; // kmer=0; // } // } return orfs; } void fillKmersSeen(byte[] bases, int[] kmersSeen, LongHashSet set, int k){ final long mask=~((-1L)<<(2*k)); long kmer=0; int len=0; int seen=0; for(int i=0; i<bases.length; i++){ final byte b=bases[i]; final int num=AminoAcid.baseToNumber[b]; if(num>=0){ len++; kmer=((kmer<<2)|num)&mask; if(len>=k && set.contains(kmer)){seen++;} }else{ len=0; } kmersSeen[i]=seen; } } boolean refineRna(Orf orf, byte[] bases, int strand, StatsContainer sc, float[] scores, int[] kmersSeen){ if(orf==null){return false;} if(verbose){System.err.println("REFINE: "+orf.toStringFlipped());} final int window=sc.lengthAvg; // final int halfWindow=window/2; // final int maxWindow=(int)(window*1.5f); final int slop=Tools.max(60, 10+window/8); final float invWindow=sc.invLengthAvg; final float oldScore=orf.orfScore; IntList starts=new IntList(); IntList stops=new IntList(); FloatList startScores=new FloatList(); FloatList stopScores=new FloatList(); final int leftmost=Tools.max(0, orf.start-slop); final int rightmost=Tools.min(bases.length-1, orf.stop+slop); if(kmersSeen!=null){ if(kmersSeen[leftmost]>=kmersSeen[rightmost]){ // System.err.println("Bad: "+oldScore); orf.orfScore=-999; return false; }else{ // System.err.println("Good: "+oldScore); } } if(verbose){System.err.println("REFINE2");} {//starts final int left=leftmost; final int right=Tools.min(bases.length-1, orf.stop+slop-window); final float thresh=cutoff3[sc.type]; fillPoints(left, right, bases, sc.start, thresh, starts, startScores); } if(verbose){System.err.println("starts: "+starts.size);} // if((orf.start+"").startsWith("146") || true){System.err.println(starts);} if(verbose){System.err.println(startScores);} {//stops final int left=Tools.max(0, orf.start-slop+window); final int right=rightmost; final float thresh=cutoff4[sc.type]; fillPoints(left, right, bases, sc.stop, thresh, stops, stopScores); } if(verbose){System.err.println("stops: "+stops.size);} // if((orf.start+"").startsWith("146") || true){System.err.println(stops);} if(verbose){System.err.println(stopScores);} final float innerThresh=cutoff5[sc.type]; final int minlen=Tools.max(window/2, window-slop); final int maxlen=window+slop; orf.orfScore=0; int lastStop=0; for(int i=0; i<starts.size; i++){ final int start=starts.get(i); final int startSeen=kmersSeen==null ? 0 : kmersSeen[start]; final float startScore=startScores.get(i); // System.err.println("start="+start); for(int j=lastStop; j<stops.size; j++){ final int stop=stops.get(j); final int rnalen=stop-start+1; // System.err.println("stop="+stop); if(rnalen<minlen){ lastStop=j+1; }else if(rnalen>maxlen){ // System.err.println("broke"); break; }else if(kmersSeen!=null && kmersSeen[stop]<=startSeen){//TODO: Test this // //skip }else{ final int len=stop-start+1; final float stopScore=stopScores.get(j); final float innerScore=(scores[stop]-scores[start])/len; // System.err.println("innerScore="+innerScore); assert(rnalen<=maxlen) : "start="+start+", stop="+stop+", rnalen="+rnalen+", minlen="+minlen+", maxlen="+maxlen; if(innerScore>=innerThresh){ final float a=Tools.max(startScore+0.25f, 0.25f); final float b=Tools.max(stopScore+0.25f, 0.25f); final float c=innerScore*innerScore; final float d=(window-(2.4f*Tools.absdif(len, window)))*invWindow; final float score=c*d*(float)Math.sqrt(a*b); if(verbose && score>=0.2f*orf.orfScore){ final int start2=(strand==0 ? start : bases.length-stop-1); final int stop2=(strand==0 ? stop : bases.length-start-1); System.err.println(start2+"-"+stop2+", "+startScore+", "+stopScore+", "+innerScore+", "+(score*scoreMult[sc.type])+", "+oldScore); } if(score>orf.orfScore){ orf.start=start; orf.stop=stop; orf.kmerScore=innerScore*len; // if(verbose){ // final int start2=(strand==0 ? start : bases.length-stop-1); // final int stop2=(strand==0 ? stop : bases.length-start-1); // System.err.println(start2+"-"+stop2+", "+startScore+", "+stopScore+", "+innerScore+", "+(score*scoreMult[sc.type])+", "+oldScore); // } orf.startScore=startScore; orf.stopScore=stopScore; orf.orfScore=score; } }else{ assert(true); } } } } orf.orfScore*=scoreMult[sc.type]; if(starts.isEmpty() || stops.isEmpty()){ if(verbose){System.err.println("No starts or stops.");} orf.orfScore=Tools.min(-999, orf.orfScore-9999); return false; } return refineByAlignment(orf, bases, strand, sc);//Returns true if no consensus is present } boolean refineByAlignment(Orf orf, byte[] bases, int strand, StatsContainer sc){ if(verbose){System.err.println("ALIGN");} Read[] consensus=ProkObject.consensusReads(sc.type); if(consensus==null || consensus.length==0){return true;} boolean refined=false; // System.err.println("Initial: "+orf.start+", "+orf.stop); for(Read r : consensus){ // refined=refineByAlignment(orf, bases, strand, sc, r.bases, 15, 15, 2); refined=refineByAlignment(orf, bases, strand, sc, r.bases, sc.startSlop(), sc.stopSlop(), 2); if(refined || sc.type==r18S || true){break;} } if(refined){ if(verbose){System.err.println("Aligned to: "+orf.start+", "+orf.stop);} }else{ if(verbose){System.err.println("Alignment failed.");} orf.orfScore=Tools.min(-999, orf.orfScore-9999); } return refined; } boolean refineByAlignment(Orf orf, byte[] bases, int strand, StatsContainer sc, byte[] consensus, final int startSlop, final int stopSlop, int recurLimit){ final int start0=orf.start; final int stop0=orf.stop; assert(start0>=0 && start0<bases.length) : start0+", "+stop0; assert(stop0>=0 && stop0<bases.length) : start0+", "+stop0; final float minID=sc.minIdentity(); final int padding=Tools.min(alignmentPadding, 30+sc.lengthAvg/4); final int a=Tools.max(0, orf.start-padding); final int b=Tools.min(bases.length-1, orf.stop+padding); final int reflen=b-a+1; if(reflen>10*sc.lengthAvg && reflen>20000){ System.err.println("Skipped reflen "+reflen+"/"+sc.lengthAvg+" for " + "seqlen="+bases.length+", orf="+orf.toString()); assert(false); //TODO: Possibly change return to -1, 0, 1 ("can't align") //Should be a limit on window size... //Also consider shrinking matrix after jumbo alignments return false; } assert(a>=0 && b<bases.length) : a+", "+b; SingleStateAlignerFlat2 ssa=getSSA(); final int minScore=ssa.minScoreByIdentity(consensus.length, minID); int[] max=ssa.fillUnlimited(consensus, bases, a, b, minScore); if(max==null){return false;} final int rows=max[0]; final int maxCol=max[1]; final int maxState=max[2]; // final int maxScore=max[3]; // final int maxStart=max[4]; //returns {score, bestRefStart, bestRefStop} //padded: {score, bestRefStart, bestRefStop, padLeft, padRight}; final int[] score=ssa.score(consensus, bases, a, b, rows, maxCol, maxState); final int rstart=Tools.max(score[1], 0); final int rstop=Tools.min(score[2], bases.length-1); final float id=ssa.tracebackIdentity(consensus, bases, a, b, rows, maxCol, maxState, null); assert(rstart>=0 && rstart<bases.length) : "bases="+bases.length+ ", rstart="+rstart+", rstop="+rstop+", a="+a+", b="+b+"\n"+"score="+Arrays.toString(score)+", id="+id; assert(rstop>=0 && rstop<bases.length) : rstart+", "+rstop; if(score.length>3 && recurLimit>0){ final int padLeft=score[3]; final int padRight=score[4]; //TODO: This takes extra memory. It may be better to cap the width or ignore start0/stop0 here. int rstart2=Tools.max(0, Tools.min(start0, rstart)-10); int rstop2=Tools.min(bases.length-1, Tools.max(stop0, rstop)+10); assert(rstart2>=0 && rstart2<bases.length) : rstart2+", "+rstop2; assert(rstop2>=0 && rstop2<bases.length) : rstart2+", "+rstop2; orf.start=rstart2; orf.stop=rstop2; if(orf.start<a || orf.stop>b){ boolean ret=refineByAlignment(orf, bases, strand, sc, consensus, 0, 0, recurLimit-1); if(ret){return true;} } orf.start=start0; orf.stop=stop0; // assert(false) : "Don't traceback after recur."; } if(verbose){ System.err.println("Identity: "+id); } // assert(score.length==3) : "TODO: Handle padding requests."; // System.err.println("Identity: "+String.format("%.2f", 100*id)+"; location: "+rstart+"-"+rstop); if(id<minID){return false;} if(Tools.absdif(rstart, start0)>startSlop){orf.start=rstart;} if(Tools.absdif(rstop, stop0)>stopSlop){orf.stop=rstop;} return true; } void fillPoints(final int left, final int right, final byte[] bases, final FrameStats fs, float thresh, final IntList points, final FloatList scores){ points.clear(); scores.clear(); final float minThresh=thresh;//thresh*0.05f; // System.err.println(left+", "+right+", "+thresh+", "+minThresh); while(points.size()<8 && thresh>=minThresh){ // System.err.println("Running with thresh="+thresh); points.clear(); scores.clear(); for(int i=left; i<right; i++){ float score=fs.scorePoint(i, bases); // System.err.println(i+", "+score); if(score>=thresh){ points.add(i); scores.add(score); } } thresh=thresh*0.75f; } } /** * Generate all possible genes from each Orf, and return them in a new set of lists. * @param frameLists * @param bases * @return Lists of orfs. */ private ArrayList<Orf>[] breakOrfs(ArrayList<Orf>[] frameLists, byte[] bases){ @SuppressWarnings("unchecked") ArrayList<Orf>[] brokenLists=new ArrayList[6]; for(int strand=0; strand<2; strand++){ for(int frame=0; frame<3; frame++){ int fnum=frame+3*strand; ArrayList<Orf> longest=frameLists[fnum]; //Longest Orf per stop ArrayList<Orf> broken=new ArrayList<Orf>(); //All high scoring Orfs, including multiple per stop, with different starts if(longest!=null) { for(Orf orf : longest){ assert(orf.frame==frame); assert(orf.strand==strand); ArrayList<Orf> temp=breakOrf(orf, bases); if(temp!=null){ broken.addAll(temp); } } } Collections.sort(broken); brokenLists[fnum]=broken; } //Reverse-complement bases after processing each strand AminoAcid.reverseComplementBasesInPlace(bases); } return brokenLists; } /** * Generate an Orf for each possible start codon. * Retain only the high-scoring ones. * @param longest Longest open reading frame for a given stop. * @param bases Bases, oriented for this Orf. * @return List of Orfs. */ private ArrayList<Orf> breakOrf(Orf longest, byte[] bases){ assert(longest.start<longest.stop); final int flipped=longest.flipped(); if(flipped==1){longest.flip();}//Now the orf is aligned to its native strand geneStopsMade++; final FrameStats innerStats=pgm.statsCDS.inner; final FrameStats startStats=pgm.statsCDS.start; final FrameStats stopStats=pgm.statsCDS.stop; final String name=longest.scafName; final int start=longest.start; final int stop=longest.stop; final int strand=longest.strand; final int max=Tools.min(longest.stop-2, longest.stop-minLen+4); assert(pgm!=null) : pgm; assert(pgm.statsCDS!=null) : pgm; assert(pgm.statsCDS.inner!=null) : pgm.statsCDS; assert(pgm.statsCDS.inner.k>0) : pgm.statsCDS.inner; final int k=innerStats.k; final int mask=~((-1)<<(2*k)); final float stopScore=stopStats.scorePoint(longest.stop, bases); stCds.geneStopScoreSum+=stopScore; stCds.geneStopScoreCount++; ArrayList<Orf> broken=new ArrayList<Orf>(); int created=0; int codon=0; int kmer=0; int len=0; int numKmers=0; float currentScore=0; for(int pos=start, currentFrame=0; pos<=stop; pos++){ final byte b=bases[pos]; final int x=AminoAcid.baseToNumber[b]; codon=((codon<<2)|x); kmer=((kmer<<2)|x)&mask; if(x>=0){ len++; if(len>=k){ float prob=innerStats.probs[currentFrame][kmer]; float dif=prob-0.99f;//Prob above 1 is more likely than average currentScore+=dif; // outstream.println("pos="+pos+"\tdif="+String.format(Locale.ROOT, "%.4f", dif)+",\tscore="+String.format(Locale.ROOT, "%.4f", currentScore)+ // "\tasStart="+String.format(Locale.ROOT, "%.4f", pgm.calcStartScore(pos-2, bases))+"\tasStop="+String.format(Locale.ROOT, "%.4f", stopStats.scorePoint(pos, bases))+ // "\tcodon="+AminoAcid.kmerToString(kmer, 3)+" frame="+(currentFrame)); }else{ // outstream.println("pos="+pos+"\tdif="+String.format(Locale.ROOT, "%.4f", 0.0)+",\tscore="+String.format(Locale.ROOT, "%.4f", 0.0)+ // "\tasStart="+String.format(Locale.ROOT, "%.4f", pgm.calcStartScore(pos-2, bases))+"\tasStop="+String.format(Locale.ROOT, "%.4f", stopStats.scorePoint(pos, bases))+ // "\tcodon="+AminoAcid.kmerToString(kmer, 3)+" frame="+(currentFrame)); } }else{ len=0; kmer=0; } currentFrame++; // outstream.println("pos="+pos+", codon="+AminoAcid.kmerToString(kmer, 3)+", frame="+currentFrame+", start="+start+", isStartCodon="+pgm.isStartCodon(codon)); if(currentFrame>2){ currentFrame=0; if(pos<max && created<breakLimit && (pos==start+2 || pgm.isStartCodon(codon))){ // outstream.println(x); int glen=stop-pos+3; assert(glen>=minLen) : "glen="+glen+", minLen="+minLen+", pos="+pos+", max="+max+", start="+start; int oStart=pos-2; float startScore=startStats.scorePoint(oStart, bases); stCds.geneStartScoreSum+=startScore; stCds.geneStartScoreCount++; stCds.lengthSum+=(stop-(pos-2)+1); stCds.lengthCount++; if((startScore>=minStartScore || pos<6) /* && stopScore>=minStopScore /*|| broken.isEmpty()*/){ Orf orf=new Orf(name, pos-2, stop, strand, longest.frame, bases, false, longest.type); geneStartsMade++; orf.kmerScore=currentScore; orf.startScore=startScore; orf.stopScore=stopScore; assert(orf.frame==longest.frame); assert(orf.strand==strand); if(strand==1){orf.flip();} broken.add(orf); created++; } } codon=0; } } final int size=broken.size(); final int sizeCutoff=Tools.max(5, size/2); if(size<1){return broken;} Orf best=broken.get(0); Orf bestStart=broken.get(0); for(int i=0; i<size; i++){ Orf orf=broken.get(i); //This fixes scores because they were generated together, from start to stop, to make this O(N) instead of O(N^2). orf.kmerScore=currentScore-orf.kmerScore; orf.orfScore=orf.calcOrfScore(); if(orf.orfScore>=best.orfScore){best=orf;} if(orf.startScore>=bestStart.startScore){bestStart=orf;} stCds.geneInnerScoreSum+=orf.averageKmerScore(); stCds.geneInnerScoreCount++; } //Sort to by score descending to eliminate low-scoring copies. if(broken.size()>1){Collections.sort(broken, Feature.featureComparatorScore);} int removed=0; for(int i=0; i<size; i++){//Fix scores because they were generated together, from start to stop, to make this O(N) instead of O(N^2). Orf orf=broken.get(i); if(orf.averageKmerScore()<minInnerScore || orf.orfScore<minOrfScore || orf.orfScore/orf.length()<minAvgScore || orf.orfScore<0.5f*best.orfScore-10 || (orf.startScore<bestStart.startScore-0.55f && orf.kmerScore<best.kmerScore*1.1f && orf!=best)){ broken.set(i, null); removed++; }else if(i>sizeCutoff){ broken.set(i, null); removed++; } } if(removed>0){ Tools.condenseStrict(broken); } geneStartsRetained+=broken.size(); geneStopsRetained+=(broken.size()>0 ? 1 : 0); if(flipped==1){longest.flip();} return broken; } /*--------------------------------------------------------------*/ /*---------------- Fields ----------------*/ /*--------------------------------------------------------------*/ /** * Current gene model. * TODO: Dynamically swap this as needed for contigs with varying GC. */ GeneModel pgm; //Gene-calling cutoffs final int minLen; final int maxOverlapSameStrand; final int maxOverlapOppositeStrand; final float minStartScore; final float minStopScore; final float minInnerScore; final float minOrfScore; final float minAvgScore; // static float[] cutoff1=new float[] {0, 40f, 200f, 150f, 45f}; // static float[] cutoff2=new float[] {0, 44f, 300f, 150f, 60f}; // static float[] cutoff3=new float[] {0, 1.7f, 1.5f, 1.4f, 1.8f}; // static float[] cutoff4=new float[] {0, 1.6f, 1.5f, 0.6f, 1.1f}; // static float[] cutoff5=new float[] {0, 1.0f, 1.0f, 1.0f, 1.55f};//inner score // static float[] scoreMult=new float[] {1f, 8f, 200f, 175f, 12f}; // static float[] biases=new float[] {1f, 1.17f, 1.2f, 1.2f, 1.15f}; // static float[] cutoff1=new float[] {0, 40f, 140f, 150f, 45f}; // static float[] cutoff2=new float[] {0, 44f, 300f, 150f, 45f}; // static float[] cutoff3=new float[] {0, 1.7f, 1.1f, 1.3f, 1.8f}; // static float[] cutoff4=new float[] {0, 1.6f, 1.3f, 0.5f, 1.1f}; // static float[] cutoff5=new float[] {0, 1.0f, 1.0f, 1.0f, 1.3f};//inner score // static float[] scoreMult=new float[] {1f, 8f, 200f, 175f, 12f}; // static float[] biases=new float[] {1f, 1.17f, 1.2f, 1.2f, 1.15f}; //for k=4,2,2 // static float[] cutoff1=new float[] {0, 40f, 140f, 150f, 40f}; // static float[] cutoff2=new float[] {0, 44f, 300f, 150f, 45f}; // static float[] cutoff3=new float[] {0, 1.7f, 1.1f, 1.1f, 1.8f}; // static float[] cutoff4=new float[] {0, 1.6f, 1.3f, 0.45f, 1.1f}; // static float[] cutoff5=new float[] {0, 1.0f, 1.0f, 1.0f, 1.3f};//inner score // static float[] scoreMult=new float[] {1f, 8f, 200f, 175f, 12f}; // static float[] biases=new float[] {1f, 1.17f, 1.2f, 1.2f, 1.22f}; // //for k=5,3,3 // static float[] cutoff1=new float[] {0, 40f, 300f, 300f, 135f};//sum score // static float[] cutoff2=new float[] {0, 44f, 300f, 400f, 100f};//orf score // static float[] cutoff3=new float[] {0, 1.7f, 1.5f, 1.5f, 3.5f};//start score // static float[] cutoff4=new float[] {0, 1.6f, 1.5f, 1.4f, 1.8f};//stop score // static float[] cutoff5=new float[] {0, 1.0f, 1.1f, 1.15f, 1.4f};//inner score // static float[] scoreMult=new float[] {1f, 8f, 80f, 120f, 5f};//score mult // static float[] biases=new float[] {1f, 1.17f, 1.2f, 1.2f, 1.22f};//bias for sum score //for k=6,3,3 - this has low scores for correct 23s stops; may try k=2 or k=4 for that end //Also, 5S seems to score low very for archaea // public static int CDS=0, tRNA=1, /*r16S=2,*/ r23S=3, r5S=4, r18S=5, r28S=6, RNA=7; // static float[] cutoff1=new float[] {0, 100f, 600f, 400f, 300f};//sum score // static float[] cutoff2=new float[] {0, 48f, 300f, 300f, 32f};//orf score // static float[] cutoff3=new float[] {0, 3.0f, 1.8f, 1.6f, 4.0f};//start score // static float[] cutoff4=new float[] {0, 2.8f, 2.0f, 0.90f, 1.9f};//stop score // static float[] cutoff5=new float[] {0, 2.8f, 1.1f, 1.55f, 1.4f};//inner score // static float[] scoreMult=new float[] {1f, 1f, 35f, 80f, 1.0f};//score mult // static float[] biases=new float[] {1f, 1.25f, 1.30f, 1.30f, 1.22f};//16S bias for sum score: 1.25 best for archs, 1.4 best for bacts //// {"CDS", "tRNA", "16S", "23S", "5S", "18S", "28S", "RNA"}; // static float[] cutoff1=new float[] {0, 90f, 300f, 400f, 270f, 300f};//sum score // static float[] cutoff2=new float[] {0, 48f, 300f, 300f, 32f, 300f};//orf score // static float[] cutoff3=new float[] {0, 2.8f, 1.65f, 1.6f, 3.8f, 1.65f};//start score // static float[] cutoff4=new float[] {0, 2.6f, 1.70f, 0.90f, 1.8f, 1.70f};//stop score // static float[] cutoff5=new float[] {0, 2.8f, 1.70f, 1.55f, 1.4f, 1.70f};//inner score // static float[] scoreMult=new float[] {1f, 1f, 35f, 80f, 1.0f, 35f};//score mult // static float[] biases=new float[] {1f, 1.50f, 1.30f, 1.30f, 1.30f, 1.30f}; //// {"CDS", "tRNA", "16S", "23S", "5S", "18S", "28S", "RNA"}; // static float[] cutoff1=new float[] {0, 90f, 300f, 400f, 90f, 300f};//sum score // static float[] cutoff2=new float[] {0, 48f, 300f, 300f, 24f, 300f};//orf score // static float[] cutoff3=new float[] {0, 2.8f, 1.65f, 1.6f, 2.0f, 1.65f};//start score // static float[] cutoff4=new float[] {0, 2.6f, 1.70f, 0.90f, 1.0f, 1.70f};//stop score // static float[] cutoff5=new float[] {0, 2.8f, 1.70f, 1.55f, 2.6f, 1.70f};//inner score // static float[] scoreMult=new float[] {1f, 1f, 35f, 80f, 1.25f, 35f};//score mult // static float[] biases=new float[] {1f, 1.50f, 1.30f, 1.30f, 1.50f, 1.30f}; //// {"CDS", "tRNA", "16S", "23S", "5S", "18S", "28S", "RNA"}; // static float[] cutoff1=new float[] {0, 90f, 300f, 400f, 100f, 300f};//sum score // static float[] cutoff2=new float[] {0, 48f, 300f, 300f, 32f, 300f};//orf score // static float[] cutoff3=new float[] {0, 2.8f, 1.65f, 1.6f, 1.8f, 1.65f};//start score // static float[] cutoff4=new float[] {0, 2.6f, 1.70f, 0.90f, 1.0f, 1.70f};//stop score // static float[] cutoff5=new float[] {0, 2.8f, 1.70f, 1.55f, 2.6f, 1.70f};//inner score // static float[] scoreMult=new float[] {1f, 1f, 35f, 80f, 1.25f, 35f};//score mult // static float[] biases=new float[] {1f, 1.50f, 1.30f, 1.30f, 1.55f, 1.30f}; // {"CDS", "tRNA", "16S", "23S", "5S", "18S", "28S", "RNA"}; static float[] cutoff1=new float[] {0, 20f, 300f, 400f, 100f, 400f};//sum score static float[] cutoff2=new float[] {0, 36f, 300f, 300f, 32f, 300f};//orf score //tRNA is very sensitive here static float[] cutoff3=new float[] {0, 2.4f, 1.65f, 1.6f, 1.8f, 1.5f};//start score static float[] cutoff4=new float[] {0, 1.5f, 1.70f, 0.90f, 1.0f, 1.1f};//stop score static float[] cutoff5=new float[] {0, 2.2f, 1.70f, 1.55f, 2.6f, 1.5f};//inner score static float[] scoreMult=new float[] {1f, 1.0f, 35f, 80f, 1.25f, 35f};//score mult static float[] biases=new float[] {1f, 1.45f, 1.30f, 1.30f, 1.55f, 1.50f}; long geneStopsMade=0; long geneStartsMade=0; long geneStartsRetained=0; long geneStopsRetained=0; long geneStartsOut=0; long tRNAOut=0; long r16SOut=0; long r23SOut=0; long r5SOut=0; long r18SOut=0; ScoreTracker stCds=new ScoreTracker(CDS); ScoreTracker stCds2=new ScoreTracker(CDS); ScoreTracker stCdsPass=new ScoreTracker(CDS); ScoreTracker sttRNA=new ScoreTracker(tRNA); ScoreTracker st16s=new ScoreTracker(r16S); ScoreTracker st23s=new ScoreTracker(r23S); ScoreTracker st5s=new ScoreTracker(r5S); ScoreTracker st18s=new ScoreTracker(r18S); ScoreTracker[] trackers=new ScoreTracker[] {stCdsPass, sttRNA, st16s, st23s, st5s, st18s}; public static SingleStateAlignerFlat2 getSSA(){ SingleStateAlignerFlat2 ssa=localSSA.get(); if(ssa==null){ synchronized(localSSA){ ssa=localSSA.get(); if(ssa==null){ ssa=new SingleStateAlignerFlat2(); localSSA.set(ssa); } } } return ssa; } public static SingleStateAlignerFlat3 getSSA3(){ SingleStateAlignerFlat3 ssa=localSSA3.get(); if(ssa==null){ synchronized(localSSA3){ ssa=localSSA3.get(); if(ssa==null){ ssa=new SingleStateAlignerFlat3(); localSSA3.set(ssa); } } } return ssa; } public static SingleStateAlignerFlatFloat getSSAF(){ SingleStateAlignerFlatFloat ssa=localSSAF.get(); if(ssa==null){ synchronized(localSSAF){ ssa=localSSAF.get(); if(ssa==null){ ssa=new SingleStateAlignerFlatFloat(); localSSAF.set(ssa); } } } return ssa; } private static ThreadLocal<SingleStateAlignerFlat2> localSSA=new ThreadLocal<SingleStateAlignerFlat2>(); private static ThreadLocal<SingleStateAlignerFlat3> localSSA3=new ThreadLocal<SingleStateAlignerFlat3>(); private static ThreadLocal<SingleStateAlignerFlatFloat> localSSAF=new ThreadLocal<SingleStateAlignerFlatFloat>(); // public static int maxAlignmentEndpointDifference=15; public static int alignmentPadding=300; public static int breakLimit=12; public static int lookbackPlus=70; public static int lookbackMinus=25; // pathScore+=p0+p1*(Tools.mid(p5*(p2+pathLength), p6*(p3-pathLength), p4)); //Operon length modifiers for same strand public static float p0=-30f; public static float p1=-0.35f; //Sensitive - higher decreases FP and TP public static float p2=4.0f;//Insensitive - higher positive decreases FP and TP public static float p3=12f; //Higher decreases FP (substantially) and TP (slightly) public static float p4=-10f; //Lower decreases FP (weakly) and TP (greatly) public static float p5=2.0f; //Insensitive - lower increases FP and TP public static float p6=2f; //Greater increases FP and TP // pathScore+=q1+Tools.mid(q2*prevLength, q3+q4*prevLength, q5); //Operon length modifiers for opposite strand public static float q1=-36f; public static float q2=-1.6f; //q2 and q4 must have opposite signs public static float q3=-12f; public static float q4=3.0f; //(Lower [even negative] decreases FP and TP) public static float q5=-40f; private static PrintStream outstream=System.err; static boolean verbose; }