]> git.donarmstrong.com Git - mothur.git/blobdiff - decalc.cpp
working on pam
[mothur.git] / decalc.cpp
index 7e4a4949c52b806d508df6bfc60edf9d13d1d876..98545f8e24a72dca332bf7571fc0e5e9e6936988 100644 (file)
@@ -8,4 +8,997 @@
  */
 
 #include "decalc.h"
+#include "chimera.h"
+#include "dist.h"
+#include "eachgapdist.h"
+#include "ignoregaps.h"
+#include "eachgapdist.h"
+
+//***************************************************************************************************************
+void DeCalculator::setMask(string ms) { 
+       try {
+               seqMask = ms; 
+               int count = 0;
+               maskMap.clear();
+               
+               if (seqMask.length() != 0) {
+                       //whereever there is a base in the mask, save that value is query and subject
+                       for (int i = 0; i < seqMask.length(); i++) {
+                               if (isalpha(seqMask[i])) {
+                                       h.insert(i);
+                                       maskMap[count] = i;
+                                       count++;
+                                       
+                               }
+                       }
+               }else {
+                       for (int i = 0; i < alignLength; i++) {   
+                               h.insert(i); 
+                               maskMap[count] = i;
+                               count++;
+                       }
+               }
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "setMask");
+               exit(1);
+       } 
+}
+//***************************************************************************************************************
+void DeCalculator::runMask(Sequence* seq) {
+       try{
+               
+               string q = seq->getAligned();
+               string tempQuery = "";
+               
+               //whereever there is a base in the mask, save that value is query and subject
+               set<int>::iterator setit;
+               for ( setit=h.begin() ; setit != h.end(); setit++ )  {
+                       tempQuery += q[*setit];
+               }
+               
+               //save masked values
+               seq->setAligned(tempQuery);
+               seq->setUnaligned(tempQuery);
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "runMask");
+               exit(1);
+       }
+}
+//***************************************************************************************************************
+//num is query's spot in querySeqs
+void DeCalculator::trimSeqs(Sequence* query, Sequence* subject, map<int, int>& trim) {
+       try {
+               
+               string q = query->getAligned();
+               string s = subject->getAligned();
+               
+               int front = 0;
+               for (int i = 0; i < q.length(); i++) {
+//cout << "query = " << q[i] << " subject = " << s[i] << endl;
+                       if (isalpha(q[i]) && isalpha(s[i])) { front = i; break;  }
+               }
+//cout << endl << endl;                
+               int back = 0;           
+               for (int i = q.length(); i >= 0; i--) {
+//cout << "query = " << q[i] << " subject = " << s[i] << endl;
+                       if (isalpha(q[i]) && isalpha(s[i])) { back = i; break;  }
+               }
+               
+               trim[front] = back;
+               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "trimSeqs");
+               exit(1);
+       }
+}
+//***************************************************************************************************************
+vector<int>  DeCalculator::findWindows(Sequence* query, int front, int back, int& size, int increment) {
+       try {
+               
+               vector<int> win; 
+               
+               int cutoff = back - front;  //back - front 
+                       
+               //if window is set to default
+               if (size == 0) {  if (cutoff > 1200) {  size = 300; }
+                                                       else{  size = (cutoff / 4); }  } 
+               else if (size > (cutoff / 4)) { 
+                               m->mothurOut("You have selected too large a window size for sequence " + query->getName() + ".  I will choose an appropriate window size."); m->mothurOutEndLine();
+                               size = (cutoff / 4); 
+               }
+       
+       /*      string seq = query->getAligned().substr(front, cutoff);
+                       
+               //count bases
+               int numBases = 0;
+               for (int l = 0; l < seq.length(); l++) {  if (isalpha(seq[l])) { numBases++; }  }
+//cout << "num Bases = " << numBases << endl;                  
+               //save start of seq
+               win.push_back(front);
+//cout << front << '\t';               
+               //move ahead increment bases at a time until all bases are in a window
+               int countBases = 0;
+               int totalBases = 0;  //used to eliminate window of blanks at end of sequence
+                       
+               seq = query->getAligned();
+               for (int m = front; m < (back - size) ; m++) {
+                               
+                       //count number of bases you see
+                       if (isalpha(seq[m])) { countBases++;  }
+                               
+                       //if you have seen enough bases to make a new window
+                       if (countBases >= increment) {
+                               //total bases is the number of bases in a window already.
+                               totalBases += countBases;
+//cout << "total bases = " << totalBases << endl;
+                               win.push_back(m);  //save spot in alignment
+//cout << m << '\t';
+                               countBases = 0;                         //reset bases you've seen in this window
+                       }
+                               
+                       //no need to continue if all your bases are in a window
+                       if (totalBases == numBases) {   break;  }
+               }
+       
+
+               //get last window if needed
+               if (totalBases < numBases) {   win.push_back(back-size);  }
+//cout << endl << endl;
+*/     
+               
+               //this follows wigeon, but we may want to consider that it chops off the end values if the sequence cannot be evenly divided into steps
+               for (int i = front; i < (back - size) ; i+=increment) {  win.push_back(i);  }
+
+
+       
+               return win;
+       
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "findWindows");
+               exit(1);
+       }
+}
+
+//***************************************************************************************************************
+vector<float> DeCalculator::calcObserved(Sequence* query, Sequence* subject, vector<int> window, int size) {
+       try {
+               
+               vector<float> temp;     
+               //int gaps = 0;         
+               for (int i = 0; i < window.size(); i++) {
+                                               
+                       string seqFrag = query->getAligned().substr(window[i], size);
+                       string seqFragsub = subject->getAligned().substr(window[i], size);
+                               
+                       int diff = 0;
+                       for (int b = 0; b < seqFrag.length(); b++) {
+                               //if at least one is a base and they are not equal
+                               if( (isalpha(seqFrag[b]) || isalpha(seqFragsub[b])) && (seqFrag[b] != seqFragsub[b]) ) { diff++; }
+                               
+                               //ignore gaps
+                               //if((!isalpha(seqFrag[b])) && (!isalpha(seqFragsub[b]))) { gaps++; }
+                       }
+               
+                       //percentage of mismatched bases
+                       float dist;
+                       
+                       //if the whole fragment is 0 distance = 0
+                       //if ((seqFrag.length()-gaps) == 0)  { dist =  0.0; }
+               
+                       //percentage of mismatched bases
+                       //else {  dist = diff / (float) (seqFrag.length()-gaps) * 100;   } 
+                       
+                       dist = diff / (float) (seqFrag.length()) * 100; 
+                       
+                       temp.push_back(dist);
+               }
+                       
+               return temp;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "calcObserved");
+               exit(1);
+       }
+}
+//***************************************************************************************************************
+float DeCalculator::calcDist(Sequence* query, Sequence* subject, int front, int back) {
+       try {
+               
+               //so you only look at the trimmed part of the sequence
+               int cutoff = back - front;  
+               int gaps = 0;
+                       
+               //from first startpoint with length back-front
+               string seqFrag = query->getAligned().substr(front, cutoff);
+               string seqFragsub = subject->getAligned().substr(front, cutoff);
+                                                                                                               
+               int diff = 0;
+               for (int b = 0; b < seqFrag.length(); b++) {
+                       //ignore gaps
+                       if((!isalpha(seqFrag[b])) && (!isalpha(seqFragsub[b]))) { gaps++; }
+                       if (seqFrag[b] != seqFragsub[b]) { diff++; }
+               }
+               
+               //if the whole fragment is 0 distance = 0
+               if ((seqFrag.length()-gaps) == 0)  { return 0.0; }
+               
+               //percentage of mismatched bases
+               float dist = diff / (float) (seqFrag.length()-gaps) * 100;       
+                               
+               return dist;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "calcDist");
+               exit(1);
+       }
+}
+
+//***************************************************************************************************************
+vector<float> DeCalculator::calcExpected(vector<float> qav, float coef) {
+       try {
+               
+               //for each window
+               vector<float> queryExpected;
+                       
+               for (int j = 0; j < qav.size(); j++) {          
+                               
+                       float expected = qav[j] * coef;
+                               
+                       queryExpected.push_back(expected);      
+               }
+                       
+               return queryExpected;
+                               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "calcExpected");
+               exit(1);
+       }
+}
+//***************************************************************************************************************
+float DeCalculator::calcDE(vector<float> obs, vector<float> exp) {
+       try {
+               
+               //for each window
+               float sum = 0.0;  //sum = sum from 1 to i of (oi-ei)^2
+               int numZeros = 0;
+               for (int j = 0; j < obs.size(); j++) {          
+                       
+                       //if (obs[j] != 0.0) {
+                               sum += ((obs[j] - exp[j]) * (obs[j] - exp[j])); 
+                       //}else {  numZeros++;   }
+                       
+               }
+                       
+               float de = sqrt((sum / (obs.size() - 1 - numZeros)));
+                       
+               return de;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "calcDE");
+               exit(1);
+       }
+}
+
+//***************************************************************************************************************
+
+vector<float> DeCalculator::calcFreq(vector<Sequence*> seqs, string filename) {
+       try {
+
+               vector<float> prob;
+               string freqfile = m->getRootName(filename) + "freq";
+               ofstream outFreq;
+               
+               m->openOutputFile(freqfile, outFreq);
+               
+               outFreq << "#" << m->getVersion() << endl;
+               
+               string length = toString(seqs.size());  //if there are 5000 seqs in the template then set precision to 3
+               int precision = length.length() - 1;
+               
+               //format output
+               outFreq.setf(ios::fixed, ios::floatfield); outFreq.setf(ios::showpoint);
+               
+               //at each position in the sequence
+               for (int i = 0; i < seqs[0]->getAligned().length(); i++) {
+                       
+                       vector<int> freq;   freq.resize(4,0);
+                       int gaps = 0;
+                       
+                       //find the frequency of each nucleotide
+                       for (int j = 0; j < seqs.size(); j++) {
+                               
+                               char value = seqs[j]->getAligned()[i];
+                               
+                               if(toupper(value) == 'A')                                                                       {       freq[0]++;      }
+                               else if(toupper(value) == 'T' || toupper(value) == 'U')         {       freq[1]++;      }
+                               else if(toupper(value) == 'G')                                                          {       freq[2]++;      }
+                               else if(toupper(value) == 'C')                                                          {       freq[3]++;      }
+                               else { gaps++; }
+                       }
+                       
+                       //find base with highest frequency
+                       int highest = 0;
+                       for (int j = 0; j < freq.size(); j++) {   if (freq[j] > highest) {  highest = freq[j];  }               }
+                       
+                       float highFreq = highest / (float) (seqs.size());       
+                       
+                       float Pi;
+                       Pi =  (highFreq - 0.25) / 0.75; 
+                       
+                       //cannot have probability less than 0.
+                       if (Pi < 0) { Pi = 0.0; }
+                       
+                       //saves this for later
+                       outFreq << setprecision(precision) << i << '\t' << highFreq << endl;
+       
+                       if (h.count(i) > 0) {
+                               prob.push_back(Pi); 
+                       }
+               }
+               
+               outFreq.close();
+               
+               return prob;
+                               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "calcFreq");
+               exit(1);
+       }
+}
+//***************************************************************************************************************
+vector<float>  DeCalculator::findQav(vector<int> window, int size, vector<float> probabilityProfile) {
+       try {
+               vector<float>  averages; 
+                               
+               //for each window find average
+               for (int i = 0; i < window.size(); i++) {
+                               
+                       float average = 0.0;
+                               
+                       //while you are in the window for this sequence
+                       int count = 0;
+                       for (int j = window[i]; j < (window[i]+size); j++) {   
+                               average += probabilityProfile[j];
+                               count++;
+                       }
+                               
+                       average = average / count;
+       
+                       //save this windows average
+                       averages.push_back(average);
+               }
+                               
+               return averages;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "findQav");
+               exit(1);
+       }
+}
+//***************************************************************************************************************
+//seqs have already been masked
+vector< vector<float> > DeCalculator::getQuantiles(vector<Sequence*> seqs, vector<int> windowSizesTemplate, int window, vector<float> probProfile, int increment, int start, int end) {
+       try {
+               vector< vector<float> > quan; 
+               
+               //percentage of mismatched pairs 1 to 100
+               quan.resize(100);
+
+               //for each sequence
+               for(int i = start; i < end; i++){
+               
+                       m->mothurOut("Processing sequence " + toString(i)); m->mothurOutEndLine();
+                       Sequence* query = new Sequence(seqs[i]->getName(), seqs[i]->getAligned());
+                       
+                       //compare to every other sequence in template
+                       for(int j = 0; j < i; j++){
+                               
+                               Sequence* subject = new Sequence(seqs[j]->getName(), seqs[j]->getAligned());
+                               
+                               if (m->control_pressed) { delete query; delete subject; return quan; }
+                               
+                               map<int, int> trim;
+                               map<int, int>::iterator it;
+                               
+                               trimSeqs(query, subject, trim);
+                               
+                               it = trim.begin();
+                               int front = it->first; int back = it->second;
+                               
+                               //reset window for each new comparison
+                               windowSizesTemplate[i] = window;
+                               
+                               vector<int> win = findWindows(query, front, back, windowSizesTemplate[i], increment);
+                               
+                               vector<float> obsi = calcObserved(query, subject, win, windowSizesTemplate[i]);
+                               
+                               vector<float> q = findQav(win, windowSizesTemplate[i], probProfile);
+                                                                       
+                               float alpha = getCoef(obsi, q);
+                                               
+                               vector<float> exp = calcExpected(q, alpha);
+                               
+                               float de = calcDE(obsi, exp);
+                                                               
+                               float dist = calcDist(query, subject, front, back); 
+       //cout << i << '\t' <<  j << '\t' << dist << '\t' << de << endl;                        
+                               dist = ceil(dist);
+                               
+                               //quanMember newScore(de, i, j);
+                               
+                               quan[dist].push_back(de);
+
+                               delete subject;
+                       }
+                       
+                       delete query;
+               }
+       
+
+               return quan;
+                                               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "getQuantiles");
+               exit(1);
+       }
+}
+//********************************************************************************************************************
+//sorts lowest to highest
+inline bool compareQuanMembers(quanMember left, quanMember right){
+       return (left.score < right.score);      
+} 
+//***************************************************************************************************************
+//this was going to be used by pintail to increase the sensitivity of the chimera detection, but it wasn't quite right.  may want to revisit in the future...
+void DeCalculator::removeObviousOutliers(vector< vector<float> >& quantiles, int num) {
+       try {
+                                               
+               for (int i = 0; i < quantiles.size(); i++) {
+                       
+                       //find mean of this quantile score
+                       sort(quantiles[i].begin(), quantiles[i].end());
+                       
+                       vector<float> temp;
+                       if (quantiles[i].size() != 0) {
+                               float high = quantiles[i][int(quantiles[i].size() * 0.99)];
+                               float low =  quantiles[i][int(quantiles[i].size() * 0.01)];
+                       
+                               //look at each value in quantiles to see if it is an outlier
+                               for (int j = 0; j < quantiles[i].size(); j++) {
+                                       //is this score between 1 and 99%
+                                       if ((quantiles[i][j] > low) && (quantiles[i][j] < high)) {
+                                               temp.push_back(quantiles[i][j]);
+                                       }
+                               }
+                       }
+                       quantiles[i] = temp;
+               }
+
+/*
+               //find contributer with most offending score related to it
+               int largestContrib = findLargestContrib(seen);
+       
+               //while you still have guys to eliminate
+               while (contributions.size() > 0) {
+               
+                       m->mothurOut("Removing scores contributed by sequence " + toString(largestContrib) + " in your template file."); m->mothurOutEndLine();
+                       
+                       //remove from quantiles all scores that were made using this largestContrib
+                       for (int i = 0; i < quantiles.size(); i++) {
+//cout << "before remove " << quantiles[i].size() << '\t';
+                               removeContrib(largestContrib, quantiles[i]);
+//cout << "after remove " << quantiles[i].size() << endl;
+                       }
+//cout << count << " largest contrib = " << largestContrib << endl;  count++;
+                       //remove from contributions all scores that were made using this largestContrib
+                       removeContrib(largestContrib, contributions);
+                       
+                       //"erase" largestContrib
+                       seen[largestContrib] = -1;
+                       
+                       //get next largestContrib
+                       largestContrib = findLargestContrib(seen);
+               }
+ABOVE IS ATTEMPT #1            
+**************************************************************************************************
+BELOW IS ATTEMPT #2            
+               vector<int> marked = returnObviousOutliers(quantiles, num);
+               vector<int> copyMarked = marked;
+               
+               //find the 99% of marked
+               sort(copyMarked.begin(), copyMarked.end());
+               int high = copyMarked[int(copyMarked.size() * 0.99)];
+cout << "high = " << high << endl;             
+               
+               for(int i = 0; i < marked.size(); i++) {
+                       if (marked[i] > high) { 
+                               m->mothurOut("Removing scores contributed by sequence " + toString(marked[i]) + " in your template file."); m->mothurOutEndLine();
+                               for (int i = 0; i < quantiles.size(); i++) {
+                                       removeContrib(marked[i], quantiles[i]);
+                               }
+                       }
+
+               }
+               
+               
+               //adjust quantiles
+               for (int i = 0; i < quantiles.size(); i++) {
+                       vector<float> temp;
+                       
+                       if (quantiles[i].size() == 0) {
+                               //in case this is not a distance found in your template files
+                               for (int g = 0; g < 6; g++) {
+                                       temp.push_back(0.0);
+                               }
+                       }else{
+                               
+                               sort(quantiles[i].begin(), quantiles[i].end(), compareQuanMembers);
+                               
+                               //save 10%
+                               temp.push_back(quantiles[i][int(quantiles[i].size() * 0.10)].score);
+                               //save 25%
+                               temp.push_back(quantiles[i][int(quantiles[i].size() * 0.25)].score);
+                               //save 50%
+                               temp.push_back(quantiles[i][int(quantiles[i].size() * 0.5)].score);
+                               //save 75%
+                               temp.push_back(quantiles[i][int(quantiles[i].size() * 0.75)].score);
+                               //save 95%
+                               temp.push_back(quantiles[i][int(quantiles[i].size() * 0.95)].score);
+                               //save 99%
+                               temp.push_back(quantiles[i][int(quantiles[i].size() * 0.99)].score);
+                               
+                       }
+                       
+                       quan[i] = temp;
+                       
+               }
+*/
+               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "removeObviousOutliers");
+               exit(1);
+       }
+}
+//***************************************************************************************************************
+//put quanMember in the vector based on how far they are from the 99% or 1%.  Biggest offenders in front.
+/*vector<quanMember> DeCalculator::sortContrib(map<quanMember*, float> quan) {
+       try{
+               
+               vector<quanMember> newQuan;
+               map<quanMember*, float>::iterator it;
+               
+               while (quan.size() > 0) {
+                       
+                        map<quanMember*, float>::iterator largest = quan.begin(); 
+                         
+                       //find biggest member
+                       for (it = quan.begin(); it != quan.end(); it++) {
+                               if (it->second > largest->second) {  largest = it;  }
+                       }
+cout << largest->second << '\t' << largest->first->score << '\t' << largest->first->member1 << '\t' << largest->first->member2 << endl;
+                       //save it 
+                       newQuan.push_back(*(largest->first));
+               
+                       //erase from quan
+                       quan.erase(largest);
+               }
+               
+               return newQuan;
+               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "sortContrib");
+               exit(1);
+       }
+}
+
+***************************************************************************************************************
+//used by removeObviousOutliers which was attempt to increase sensitivity of chimera detection...not currently used...
+int DeCalculator::findLargestContrib(vector<int> seen) {
+       try{
+               
+               int largest = 0;
+               
+               int largestContribs;
+               
+               for (int i = 0; i < seen.size(); i++)  {  
+                       
+                       if (seen[i] > largest) {
+                               largestContribs = i;
+                               largest = seen[i];
+                       }
+               }
+               
+               return largestContribs;
+               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "findLargestContribs");
+               exit(1);
+       }
+}
+***************************************************************************************************************
+void DeCalculator::removeContrib(int bad, vector<quanMember>& quan) {
+       try{
+       
+               vector<quanMember> newQuan;
+               for (int i = 0; i < quan.size(); i++)  {  
+                       if ((quan[i].member1 != bad) && (quan[i].member2 != bad) ) {  
+                               newQuan.push_back(quan[i]);  
+                       }
+               }
+               
+               quan = newQuan;
+               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "removeContrib");
+               exit(1);
+       }
+}
+*/
+//***************************************************************************************************************
+float DeCalculator::findAverage(vector<float> myVector) {
+       try{
+               
+               float total = 0.0;
+               for (int i = 0; i < myVector.size(); i++)  {  total += myVector[i];  }
+               
+               float average = total / (float) myVector.size();
+               
+               return average;
+               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "findAverage");
+               exit(1);
+       }
+}
+
+//***************************************************************************************************************
+float DeCalculator::getCoef(vector<float> obs, vector<float> qav) {
+       try {
+       
+               //find average prob for this seqs windows
+               float probAverage = findAverage(qav);
+                               
+               //find observed average 
+               float obsAverage = findAverage(obs);
+                       
+               float coef = obsAverage / probAverage;
+                                               
+               return coef;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "getCoef");
+               exit(1);
+       }
+}
+//***************************************************************************************************************
+//gets closest matches to each end, since chimeras will most likely have different parents on each end
+vector<Sequence> DeCalculator::findClosest(Sequence querySeq, vector<Sequence*>& thisTemplate, vector<Sequence*>& thisFilteredTemplate, int numWanted, int minSim) {
+       try {
+               //indexes.clear();
+               
+               vector<Sequence> seqsMatches;  
+               
+               vector<SeqDist> distsLeft;
+               vector<SeqDist> distsRight;
+               
+               Dist* distcalculator = new eachGapDist();
+               
+               string queryUnAligned = querySeq.getUnaligned();
+               int numBases = int(queryUnAligned.length() * 0.33);
+               
+               string leftQuery = ""; //first 1/3 of the sequence
+               string rightQuery = ""; //last 1/3 of the sequence
+               string queryAligned = querySeq.getAligned();
+               
+               //left side
+               bool foundFirstBase = false;
+               int baseCount = 0;
+               int leftSpot = 0;
+               int firstBaseSpot = 0;
+               for (int i = 0; i < queryAligned.length(); i++) {
+                       //if you are a base
+                       if (isalpha(queryAligned[i])) {         
+                               baseCount++; 
+                               if (!foundFirstBase) {   foundFirstBase = true;  firstBaseSpot = i;  }
+                       }
+                       
+                       //eliminate opening .'s
+                       if (foundFirstBase) {   leftQuery += queryAligned[i];  }
+                       //if you have 1/3
+                       if (baseCount >= numBases) {  leftSpot = i; break; } //first 1/3
+               }
+               
+               //right side - count through another 1/3, so you are at last third
+               baseCount = 0;
+               int rightSpot = 0;
+               for (int i = leftSpot; i < queryAligned.length(); i++) {
+                       //if you are a base
+                       if (isalpha(queryAligned[i])) {         baseCount++;    }
+                       //if you have 1/3
+                       if (baseCount > numBases + 1) { rightSpot = i;  break; } //last 1/3
+               }
+               
+               //trim end
+               //find last position in query that is a non gap character
+               int lastBaseSpot = queryAligned.length()-1;
+               for (int j = queryAligned.length()-1; j >= 0; j--) {
+                       if (isalpha(queryAligned[j])) {
+                               lastBaseSpot = j;
+                               break;
+                       }
+               }
+               rightQuery = queryAligned.substr(rightSpot, (lastBaseSpot-rightSpot+1)); //sequence from pos spot to end
+               
+               Sequence queryLeft(querySeq.getName(), leftQuery);
+               Sequence queryRight(querySeq.getName(), rightQuery);
+               
+//cout << querySeq->getName() << '\t' << leftSpot << '\t' << rightSpot << '\t' << firstBaseSpot << '\t' << lastBaseSpot << endl;
+//cout << queryUnAligned.length() << '\t' << queryLeft.getUnaligned().length() << '\t' << queryRight.getUnaligned().length() << endl;
+               for(int j = 0; j < thisFilteredTemplate.size(); j++){
+                       
+                       string dbAligned = thisFilteredTemplate[j]->getAligned();
+                       string leftDB = dbAligned.substr(firstBaseSpot, (leftSpot-firstBaseSpot+1)); //first 1/3 of the sequence
+                       string rightDB = dbAligned.substr(rightSpot, (lastBaseSpot-rightSpot+1)); //last 1/3 of the sequence
+
+                       Sequence dbLeft(thisFilteredTemplate[j]->getName(), leftDB);
+                       Sequence dbRight(thisFilteredTemplate[j]->getName(), rightDB);
+
+                       distcalculator->calcDist(queryLeft, dbLeft);
+                       float distLeft = distcalculator->getDist();
+                       
+                       distcalculator->calcDist(queryRight, dbRight);
+                       float distRight = distcalculator->getDist();
+
+                       SeqDist subjectLeft;
+                       subjectLeft.seq = NULL;
+                       subjectLeft.dist = distLeft;
+                       subjectLeft.index = j;
+                       
+                       distsLeft.push_back(subjectLeft);
+                       
+                       SeqDist subjectRight;
+                       subjectRight.seq = NULL;
+                       subjectRight.dist = distRight;
+                       subjectRight.index = j;
+                       
+                       distsRight.push_back(subjectRight);
+
+               }
+               
+               delete distcalculator;
+               
+               //sort by smallest distance
+               sort(distsRight.begin(), distsRight.end(), compareSeqDist);
+               sort(distsLeft.begin(), distsLeft.end(), compareSeqDist);
+
+               
+               //merge results         
+               map<string, string> seen;
+               map<string, string>::iterator it;
+               
+               vector<SeqDist> dists;
+               float lastRight = distsRight[0].dist;
+               float lastLeft = distsLeft[0].dist;
+
+               float maxDist = 1.0 - (minSim / 100.0);
+
+               for (int i = 0; i < numWanted+1; i++) {
+                       if (m->control_pressed) { return seqsMatches; }
+                       
+                       //add left if you havent already
+                       it = seen.find(thisTemplate[distsLeft[i].index]->getName());
+                       if (it == seen.end() && distsLeft[i].dist <= maxDist) {  
+                               dists.push_back(distsLeft[i]);
+                               seen[thisTemplate[distsLeft[i].index]->getName()] = thisTemplate[distsLeft[i].index]->getName();
+                               lastLeft =  distsLeft[i].dist;
+//                             cout << "loop-left\t" << db[distsLeft[i].index]->getName() << '\t' << distsLeft[i].dist << endl;
+                       }
+
+                       //add right if you havent already
+                       it = seen.find(thisTemplate[distsRight[i].index]->getName());
+                       if (it == seen.end() && distsRight[i].dist <= maxDist) {  
+                               dists.push_back(distsRight[i]);
+                               seen[thisTemplate[distsRight[i].index]->getName()] = thisTemplate[distsRight[i].index]->getName();
+                               lastRight =  distsRight[i].dist;
+//                             cout << "loop-right\t" << db[distsRight[i].index]->getName() << '\t' << distsRight[i].dist << endl;
+                       }
+                       
+                       if (i == numWanted) { break; }
+                       
+               }
+               
+               //are we still above the minimum similarity cutoff
+               if ((lastLeft >= minSim) || (lastRight >= minSim)) {
+                       //add in ties from left
+                       int i = numWanted;
+                       while (i < distsLeft.size()) {  
+                               if (distsLeft[i].dist == lastLeft) {  dists.push_back(distsLeft[i]);  }
+                               else { break; }
+                               i++;
+                       }
+                       
+                       //add in ties from right
+                       i = numWanted;
+                       while (i < distsRight.size()) {  
+                               if (distsRight[i].dist == lastRight) {  dists.push_back(distsRight[i]);  }
+                               else { break; }
+                               i++;
+                       }
+               }
+               
+               //cout << numWanted << endl;
+               for (int i = 0; i < dists.size(); i++) {
+//                     cout << db[dists[i].index]->getName() << '\t' << dists[i].dist << endl;
+
+                       if ((thisTemplate[dists[i].index]->getName() != querySeq.getName()) && (((1.0-dists[i].dist)*100) >= minSim)) {
+                               Sequence temp(thisTemplate[dists[i].index]->getName(), thisTemplate[dists[i].index]->getAligned()); //have to make a copy so you can trim and filter without stepping on eachother.
+                               //cout << querySeq->getName() << '\t' << thisTemplate[dists[i].index]->getName()  << '\t' << dists[i].dist << endl;
+                               seqsMatches.push_back(temp);
+                       }
+
+               }
+               
+               return seqsMatches;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "findClosest");
+               exit(1);
+       }
+}
+//***************************************************************************************************************
+Sequence* DeCalculator::findClosest(Sequence* querySeq, vector<Sequence*> db) {
+       try {
+               
+               Sequence* seqsMatch;  
+               
+               Dist* distcalculator = new eachGapDist();
+               int index = 0;
+               int smallest = 1000000;
+               
+               for(int j = 0; j < db.size(); j++){
+                       
+                       distcalculator->calcDist(*querySeq, *db[j]);
+                       float dist = distcalculator->getDist();
+                       
+                       if (dist < smallest) { 
+                               smallest = dist;
+                               index = j;
+                       }
+               }
+               
+               delete distcalculator;
+               
+               seqsMatch = new Sequence(db[index]->getName(), db[index]->getAligned()); //have to make a copy so you can trim and filter without stepping on eachother.
+                       
+               return seqsMatch;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "findClosest");
+               exit(1);
+       }
+}
+/***************************************************************************************************************/
+map<int, int> DeCalculator::trimSeqs(Sequence& query, vector<Sequence>& topMatches) {
+       try {
+               
+               int frontPos = 0;  //should contain first position in all seqs that is not a gap character
+               int rearPos = query.getAligned().length();
+               
+               //********find first position in topMatches that is a non gap character***********//
+               //find first position all query seqs that is a non gap character
+               for (int i = 0; i < topMatches.size(); i++) {
+                       
+                       string aligned = topMatches[i].getAligned();
+                       int pos = 0;
+                       
+                       //find first spot in this seq
+                       for (int j = 0; j < aligned.length(); j++) {
+                               if (isalpha(aligned[j])) {
+                                       pos = j;
+                                       break;
+                               }
+                       }
+                       
+                       //save this spot if it is the farthest
+                       if (pos > frontPos) { frontPos = pos; }
+               }
+               
+               
+               string aligned = query.getAligned();
+               int pos = 0;
+                       
+               //find first position in query that is a non gap character
+               for (int j = 0; j < aligned.length(); j++) {
+                       if (isalpha(aligned[j])) {
+                               pos = j;
+                               break;
+                       }
+               }
+               
+               //save this spot if it is the farthest
+               if (pos > frontPos) { frontPos = pos; }
+               
+               
+               //********find last position in topMatches that is a non gap character***********//
+               for (int i = 0; i < topMatches.size(); i++) {
+                       
+                       string aligned = topMatches[i].getAligned();
+                       int pos = aligned.length();
+                       
+                       //find first spot in this seq
+                       for (int j = aligned.length()-1; j >= 0; j--) {
+                               if (isalpha(aligned[j])) {
+                                       pos = j;
+                                       break;
+                               }
+                       }
+                       
+                       //save this spot if it is the farthest
+                       if (pos < rearPos) { rearPos = pos; }
+               }
+               
+               
+               aligned = query.getAligned();
+               pos = aligned.length();
+               
+               //find last position in query that is a non gap character
+               for (int j = aligned.length()-1; j >= 0; j--) {
+                       if (isalpha(aligned[j])) {
+                               pos = j;
+                               break;
+                       }
+               }
+               
+               //save this spot if it is the farthest
+               if (pos < rearPos) { rearPos = pos; }
+               
+               map<int, int> trimmedPos;
+               //check to make sure that is not whole seq
+               if ((rearPos - frontPos - 1) <= 0) {  
+                       query.setAligned("");
+                       //trim topMatches
+                       for (int i = 0; i < topMatches.size(); i++) {
+                               topMatches[i].setAligned("");
+                       }
+                       
+               }else {
+
+                       //trim query
+                       string newAligned = query.getAligned();
+                       newAligned = newAligned.substr(frontPos, (rearPos-frontPos+1));
+                       query.setAligned(newAligned);
+                       
+                       //trim topMatches
+                       for (int i = 0; i < topMatches.size(); i++) {
+                               newAligned = topMatches[i].getAligned();
+                               newAligned = newAligned.substr(frontPos, (rearPos-frontPos+1));
+                               topMatches[i].setAligned(newAligned);
+                       }
+                       
+                       for (int i = 0; i < newAligned.length(); i++) {
+                               trimmedPos[i] = i+frontPos;
+                       }
+               }
+               return trimmedPos;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "DeCalculator", "trimSequences");
+               exit(1);
+       }
+
+}
+//***************************************************************************************************************
+
+