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*
* created: 2009
*
* This file is part of Artemis
*
* Copyright(C) 2009 Genome Research Limited
*
* This program is free software; you can redistribute it and/or
* modify it under the terms of the GNU General Public License
* as published by the Free Software Foundation; either version 2
* of the License, or(at your option) any later version.
*
* This program is distributed in the hope that it will be useful,
* but WITHOUT ANY WARRANTY; without even the implied warranty of
* MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the
* GNU General Public License for more details.
*
* You should have received a copy of the GNU General Public License
* along with this program; if not, write to the Free Software
* Foundation, Inc., 59 Temple Place - Suite 330, Boston, MA 02111-1307, USA.
*
*/
import java.awt.AlphaComposite;
import java.awt.BasicStroke;
import java.awt.BorderLayout;
import java.awt.Color;
import java.awt.Component;
import java.awt.Composite;
import java.awt.GridBagConstraints;
import java.awt.GridBagLayout;
import java.awt.event.ActionEvent;
import java.awt.event.ActionListener;
import java.awt.event.AdjustmentEvent;
import java.awt.event.AdjustmentListener;
import java.awt.event.ItemEvent;
import java.awt.event.ItemListener;
import java.awt.event.KeyAdapter;
import java.awt.event.KeyEvent;
import java.awt.image.BufferedImage;
import java.io.FileOutputStream;
import java.io.IOException;
import java.lang.management.ManagementFactory;
import java.lang.management.MemoryMXBean;
import javax.swing.ButtonGroup;
import javax.swing.JLabel;
import javax.swing.JMenu;
import javax.swing.JMenuItem;
import javax.swing.JRadioButton;
import javax.swing.JSeparator;
import javax.swing.border.Border;
import javax.swing.border.EmptyBorder;
import javax.swing.text.JTextComponent;
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import org.apache.log4j.Level;
import net.sf.picard.reference.ReferenceSequenceFile;
import net.sf.picard.sam.BuildBamIndex;
import net.sf.samtools.AlignmentBlock;
import net.sf.samtools.SAMException;
import net.sf.samtools.SAMFileHeader;
import net.sf.samtools.SAMFileReader;
import net.sf.samtools.SAMRecord;
import net.sf.samtools.SAMSequenceRecord;
import net.sf.samtools.SAMFileReader.ValidationStringency;
import net.sf.samtools.util.CloseableIterator;
import uk.ac.sanger.artemis.Entry;
import uk.ac.sanger.artemis.EntryGroup;
import uk.ac.sanger.artemis.FeatureVector;
import uk.ac.sanger.artemis.Selection;
import uk.ac.sanger.artemis.SelectionChangeEvent;
import uk.ac.sanger.artemis.SelectionChangeListener;
import uk.ac.sanger.artemis.circular.TextFieldInt;
import uk.ac.sanger.artemis.components.DisplayAdjustmentEvent;
import uk.ac.sanger.artemis.components.DisplayAdjustmentListener;
import uk.ac.sanger.artemis.components.EntryEdit;
import uk.ac.sanger.artemis.components.FeatureDisplay;
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import uk.ac.sanger.artemis.components.FileViewer;
import uk.ac.sanger.artemis.components.NonModalDialog;
import uk.ac.sanger.artemis.components.SwingWorker;
import uk.ac.sanger.artemis.components.genebuilder.AutoCompleteComboDocument;
import uk.ac.sanger.artemis.editor.MultiLineToolTipUI;
import uk.ac.sanger.artemis.sequence.MarkerRange;
import uk.ac.sanger.artemis.sequence.NoSequenceException;
import uk.ac.sanger.artemis.util.Document;
import uk.ac.sanger.artemis.util.DocumentFactory;
import uk.ac.sanger.artemis.util.FTPSeekableStream;
public class BamView extends JPanel
implements DisplayAdjustmentListener, SelectionChangeListener
private Hashtable<String, SAMFileReader> samFileReaderHash = new Hashtable<String, SAMFileReader>();
private HashMap<String, Integer> seqLengths = new HashMap<String, Integer>();
private HashMap<String, Integer> offsetLengths;
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protected List<String> bamList;
protected List<Integer> hideBamList = new Vector<Integer>();
private boolean isSingle = false;
private boolean isSNPs = false;
private FeatureDisplay feature_display;
private Selection selection;
private Ruler ruler;
private int startBase = -1;
private int endBase = -1;
private boolean showBaseAlignment = false;
private JMenu bamFilesMenu = new JMenu("BAM files");
private JCheckBoxMenuItem logMenuItem = new JCheckBoxMenuItem("Use Log Scale", logScale);
private JCheckBoxMenuItem cbStackView = new JCheckBoxMenuItem("Stack", true);
private JCheckBoxMenuItem cbPairedStackView = new JCheckBoxMenuItem("Paired Stack");
private JCheckBoxMenuItem cbStrandStackView = new JCheckBoxMenuItem("Strand Stack");
private JCheckBoxMenuItem cbIsizeStackView = new JCheckBoxMenuItem("Inferred Size", false);
private JCheckBoxMenuItem cbCoverageView = new JCheckBoxMenuItem("Coverage", false);
private JCheckBoxMenuItem cbCoverageStrandView = new JCheckBoxMenuItem("Coverage by Strand", false);
private JCheckBoxMenuItem cbLastSelected;
private ButtonGroup buttonGroup = new ButtonGroup();
private JCheckBoxMenuItem colourByCoverageColour = new JCheckBoxMenuItem("Coverage Plot Colours");
private JCheckBoxMenuItem baseQualityColour = new JCheckBoxMenuItem("Base Quality");
private JCheckBoxMenuItem markInsertions = new JCheckBoxMenuItem("Mark Insertions", true);
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private AlphaComposite translucent =
AlphaComposite.getInstance(AlphaComposite.SRC_OVER, 0.6f);
private CoveragePanel coverageView = new CoveragePanel();
protected static String BAM_SUFFIX = ".*\\.(bam|cram)$";
private static Color LIGHT_GREY = new Color(200, 200, 200);
private static Color DARK_GREEN = new Color(0, 150, 0);
private static Color DARK_ORANGE = new Color(255,140,0);
private static Color DEEP_PINK = new Color(139,10,80);
private Point lastMousePoint = null;
private SAMRecord mouseOverSAMRecord = null;
private SAMRecord highlightSAMRecord = null;
private String mouseOverInsertion;
// record of where a mouse drag starts
private static int MAX_BASES = 26000;
private int maxHeight = 800;
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private int BASE_HEIGHT;
private PopupMessageFrame popFrame = new PopupMessageFrame();
private PopupMessageFrame waitingFrame = new PopupMessageFrame("waiting...");
public static org.apache.log4j.Logger logger4j =
org.apache.log4j.Logger.getLogger(BamView.class);
public BamView(List<String> bamList,
String reference,
int nbasesInView,
final EntryEdit entry_edit,
final FeatureDisplay feature_display,
final Bases bases,
final JPanel containerPanel,
final JFrame frame)
{
this(bamList, reference, nbasesInView, feature_display, bases, containerPanel, frame);
this.entry_edit = entry_edit;
}
public BamView(List<String> bamList,
int nbasesInView,
final FeatureDisplay feature_display,
final Bases bases,
this.bamList = bamList;
this.feature_display = feature_display;
this.bases = bases;
containerPanel.setLayout(new BoxLayout(containerPanel, BoxLayout.Y_AXIS));
containerPanel.add(mainPanel);
// filter out unmapped reads by default
setSamRecordFlagPredicate(
new SAMRecordFlagPredicate(SAMRecordFlagPredicate.READ_UNMAPPED_FLAG));
System.setProperty("reference", reference); // for CRAM
try
{
getEntry(reference,entryGroup);
}
catch (NoSequenceException e)
{
e.printStackTrace();
}
}
try
{
readHeaderPicard();
}
catch(java.lang.UnsupportedClassVersionError err)
{
JOptionPane.showMessageDialog(null,
"This requires Java 1.6 or higher.",
"Check Java Version", JOptionPane.WARNING_MESSAGE);
}
catch (IOException e)
{
e.printStackTrace();
}
final javax.swing.plaf.FontUIResource font_ui_resource =
Options.getOptions().getFontUIResource();
while(keys.hasMoreElements())
{
Object key = keys.nextElement();
Object value = UIManager.get(key);
if(value instanceof javax.swing.plaf.FontUIResource)
UIManager.put(key, font_ui_resource);
}
setFont(Options.getOptions().getFont());
ALIGNMENT_PIX_PER_BASE = fm.charWidth('M');
BASE_HEIGHT = fm.getMaxAscent();
MultiLineToolTipUI.initialize();
setToolTipText("");
buttonGroup.add(cbStackView);
buttonGroup.add(cbPairedStackView);
buttonGroup.add(cbStrandStackView);
buttonGroup.add(cbIsizeStackView);
buttonGroup.add(cbCoverageView);
buttonGroup.add(cbCoverageStrandView);
jspView = new JScrollPane(this,
JScrollPane.VERTICAL_SCROLLBAR_AS_NEEDED,
JScrollPane.HORIZONTAL_SCROLLBAR_NEVER);
jspView.setViewportBorder(BorderFactory.createMatteBorder(0, 0, 1, 0, Color.DARK_GRAY));
Border empty = new EmptyBorder(0,0,0,0);
jspView.setBorder(empty);
jspView.getVerticalScrollBar().setUnitIncrement(8);
}
public String getToolTipText()
{
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if(mouseOverSAMRecord == null)
return null;
String msg =
mouseOverSAMRecord.getReadName() + "\n" +
mouseOverSAMRecord.getAlignmentStart() + ".." +
mouseOverSAMRecord.getAlignmentEnd() + "\nisize=" +
mouseOverSAMRecord.getInferredInsertSize() + "\nmapq=" +
mouseOverSAMRecord.getMappingQuality()+"\nrname="+
mouseOverSAMRecord.getReferenceName();
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if( mouseOverSAMRecord.getReadPairedFlag() &&
mouseOverSAMRecord.getProperPairFlag() &&
!mouseOverSAMRecord.getMateUnmappedFlag())
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{
msg = msg +
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(mouseOverSAMRecord.getReadNegativeStrandFlag() ? "-" : "+")+" / "+
(mouseOverSAMRecord.getMateNegativeStrandFlag() ? "-" : "+");
}
else
msg = msg +
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(mouseOverSAMRecord.getReadNegativeStrandFlag() ? "-" : "+");
if(msg != null && mouseOverInsertion != null)
msg = msg + "\nInsertion at:" +mouseOverInsertion;
return msg;
/**
* Get the BAM index file from the list
* @param bam
* @return
* @throws IOException
*/
private File getBamIndexFile(String bam) throws IOException
{
File bamIndexFile = null;
if (bam.startsWith("http") || bam.startsWith("ftp"))
{
final URL urlBamIndexFile = new URL(bam + ".bai");
InputStream is = urlBamIndexFile.openStream();
// Create temp file.
bamIndexFile = File.createTempFile(urlBamIndexFile.getFile().replaceAll(
"[\\/\\s]", "_"), ".bai");
bamIndexFile.deleteOnExit();
FileOutputStream out = new FileOutputStream(bamIndexFile);
int c;
while ((c = is.read()) != -1)
out.write(c);
out.flush();
out.close();
is.close();
logger4j.debug("create... " + bamIndexFile.getAbsolutePath());
{
final File cramIndexFile = new File(bam + ".crai");
if(cramIndexFile.exists())
return cramIndexFile;
}
/**
* Get the SAM file reader.
* @param bam
* @return
* @throws IOException
*/
private SAMFileReader getSAMFileReader(final String bam) throws IOException
{
if(samFileReaderHash.containsKey(bam))
return samFileReaderHash.get(bam);
if(!bamIndexFile.exists())
{
try
{
logger4j.warn("Index file not found so using picard to index the BAM.");
// Use Picard to index the file
// requires reads to be sorted by coordinate
new BuildBamIndex().instanceMain(
new String[]{ "I="+bam, "O="+bamIndexFile.getAbsolutePath(), "MAX_RECORDS_IN_RAM=50000", "VALIDATION_STRINGENCY=SILENT" });
}
catch(SAMException e)
{
String ls = System.getProperty("line.separator");
String msg =
"BAM index file is missing. The BAM file needs to be sorted and indexed"+ls+
"This can be done using samtools (http://samtools.sf.net/):"+ls+ls+
"samtools sort <in.bam> <out.prefix>"+ls+
"samtools index <sorted.bam>";
throw new SAMException(msg);
}
if(feature_display != null && bam.endsWith("cram"))
{
final CRAMReferenceSequenceFile ref = new CRAMReferenceSequenceFile(
feature_display.getEntryGroup().getSequenceEntry(), this);
final Map<Object, ReferenceSequenceFile> referenceFactory =
new HashMap<Object, ReferenceSequenceFile>();
referenceFactory.put(bamIndexFile, ref);
try
{
Class<?> cls = getClass().getClassLoader().loadClass("net.sf.samtools.ReferenceDiscovery");
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Field f = cls.getDeclaredField("referenceFactory");
f.set(null, referenceFactory);
}
catch (ClassNotFoundException e)
{
System.err.println("Check cramtools.jar is in the CLASSPATH. "+e.getMessage());
}
catch (SecurityException e)
{
e.printStackTrace();
}
catch (NoSuchFieldException e)
{
e.printStackTrace();
}
catch (IllegalArgumentException e)
{
e.printStackTrace();
}
catch (IllegalAccessException e)
{
e.printStackTrace();
}
//net.sf.samtools.ReferenceDiscovery.referenceFactory.put(bamIndexFile, ref);
}
if(bam.startsWith("ftp"))
{
FTPSeekableStream fss = new FTPSeekableStream(new URL(bam));
samFileReader = new SAMFileReader(fss, bamIndexFile, false);
}
else if(!bam.startsWith("http"))
samFileReader = new SAMFileReader(bamFile, bamIndexFile);
}
else
{
final URL urlBamFile = new URL(bam);
samFileReader = new SAMFileReader(urlBamFile, bamIndexFile, false);
samFileReader.setValidationStringency(ValidationStringency.SILENT);
samFileReaderHash.put(bam, samFileReader);
return samFileReader;
String bam = bamList.get(0);
final SAMFileReader inputSam = getSAMFileReader(bam);
//final SAMFileReader inputSam = new SAMFileReader(bamFile, indexFile);
SAMFileHeader header = inputSam.getFileHeader();
List<SAMSequenceRecord> readGroups = header.getSequenceDictionary().getSequences();
for(int i=0; i<readGroups.size(); i++)
{
seqLengths.put(readGroups.get(i).getSequenceName(),
readGroups.get(i).getSequenceLength());
seqNames.add(readGroups.get(i).getSequenceName());
}
private void readFromBamPicard(int start, int end, int bamIndex, float pixPerBase)
{
// Open the input file. Automatically detects whether input is SAM or BAM
// and delegates to a reader implementation for the appropriate format.
String bam = bamList.get(bamIndex);
final SAMFileReader inputSam = getSAMFileReader(bam);
//final SAMFileReader inputSam = new SAMFileReader(bamFile, indexFile);
int sLen = seqLengths.get(seqNames.get(i));
int offset = getSequenceOffset(seqNames.get(i));
int sBeg = offset+1;
int sEnd = sBeg+sLen-1;
if( (sBeg >= start && sBeg <= end) ||
(sBeg <= start && sEnd >= start) )
{
int thisStart = start - offset;
if(thisStart < 1)
thisStart = 1;
int thisEnd = end - offset;
if(thisEnd > sLen)
thisEnd = sLen;
iterateOverBam(inputSam, seqNames.get(i), thisStart, thisEnd, bamIndex, pixPerBase, bam);
//System.out.println("READ "+seqNames.get(i)+" "+thisStart+".."+thisEnd+" "+start+" --- "+offset);
}
}
}
else
{
String refName = (String) combo.getSelectedItem();
iterateOverBam(inputSam, refName, start, end, bamIndex, pixPerBase, bam);
}
/**
* Iterate over BAM file and load into the <code>List</code> of
* <code>SAMRecord</code>.
* @param inputSam
* @param refName
* @param start
* @param end
*/
private void iterateOverBam(final SAMFileReader inputSam,
String refName, int start, int end,
int bamIndex, float pixPerBase,
String bam)
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boolean multipleBAM = false;
if(bamList.size() > 1)
multipleBAM = true;
CloseableIterator<SAMRecord> it = inputSam.queryOverlapping(refName, start, end);
MemoryMXBean memory = ManagementFactory.getMemoryMXBean();
int seqOffset = getSequenceOffset(refName);
int offset = seqOffset- getBaseAtStartOfView();
if( samRecordFlagPredicate == null ||
!samRecordFlagPredicate.testPredicate(samRecord))
{
if(samRecordMapQPredicate == null ||
samRecordMapQPredicate.testPredicate(samRecord))
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if(multipleBAM)
samRecord.setAttribute("FL", bamIndex);
if(isCoverageView(pixPerBase))
coverageView.addRecord(samRecord, offset, bam);
if(isCoverage)
coveragePanel.addRecord(samRecord, offset, bam);
snpPanel.addRecord(samRecord, seqOffset);
if(!isCoverageView(pixPerBase))
readsInView.add(samRecord);
}
cnt = 0;
float heapFraction =
(float)((float)memory.getHeapMemoryUsage().getUsed()/
(float)memory.getHeapMemoryUsage().getMax());
logger4j.debug("Heap memory usage (used/max): "+heapFraction);
if(readsInView.size() > checkMemAfter*2 && !waitingFrame.isVisible())
waitingFrame.showWaiting("loading...", mainPanel);
if(heapFraction > 0.90)
"Using > 90 % of the maximum memory limit:"+
(memory.getHeapMemoryUsage().getMax()/1000000.f)+" Mb.\n"+
"Not all reads in this range have been read in. Zoom in or\n"+
"consider increasing the memory for this application.",
System.err.println(e.getMessage());
it.close();
}
private int getSequenceLength()
{
{
int len = 0;
for(int i=0; i<seqNames.size(); i++)
len += seqLengths.get(seqNames.get(i));
return len;
}
else
return seqLengths.get((String) combo.getSelectedItem());
}
/**
* For BAM files with multiple references sequences, calculate
* the offset from the start of the concatenated sequence for
* a given reference.
* @param refName
* @return
*/
protected int getSequenceOffset(String refName)
{
if(offsetLengths == null)
{
/* offsetLengths = new Hashtable<String, Integer>(combo.getItemCount());
int offset = 0;
for(int i=0; i<combo.getItemCount(); i++)
{
String thisSeqName = (String) combo.getItemAt(i);
offsetLengths.put(thisSeqName, offset);
offset += seqLengths.get(combo.getItemAt(i));
final FeatureVector features = feature_display.getEntryGroup().getAllFeatures();
final HashMap<String, Integer> lookup = new HashMap<String, Integer>();
for(int i=0; i<features.size(); i++)
lookup.put(features.elementAt(i).getIDString(), features.elementAt(i).getFirstBase());
offsetLengths = new HashMap<String, Integer>(seqNames.size());
for(int i=0; i<seqNames.size(); i++)
{
final Integer pos = lookup.get(seqNames.get(i));
if(pos != null)
offsetLengths.put(seqNames.get(i), pos-1);
/*final FeatureContigPredicate predicate = new FeatureContigPredicate(seqNames.get(i).trim());
for(int j=0; j<features.size(); j++)
{
if(predicate.testPredicate(features.elementAt(j)))
{
offsetLengths.put(seqNames.get(i), features.elementAt(j).getFirstBase()-1);
break;
}
if(offsetLengths.size() != seqNames.size())
System.err.println("Found: "+offsetLengths.size() +" of "+ seqNames.size());
SwingUtilities.invokeLater(new Runnable()
{
public void run()
{
JOptionPane.showMessageDialog(BamView.this,
"There is a problem matching the reference sequences\n"+
"to the names in the BAM file. This may mean the labels\n"+
"on the reference features do not match those in the in\n"+
"the BAM file.",
"Problem Found", JOptionPane.WARNING_MESSAGE);
}
});
int offset = 0;
for(int i=0; i<combo.getItemCount(); i++)
{
String thisSeqName = (String) combo.getItemAt(i);
offsetLengths.put(thisSeqName, offset);
offset += seqLengths.get(combo.getItemAt(i));
}
//return 0;
return offsetLengths.get(refName);
protected void paintComponent(Graphics g)
{
super.paintComponent(g);
Graphics2D g2 = (Graphics2D)g;
if(startBase > 0)
start = startBase;
else
if(endBase > 0)
end = endBase;
else
end = start + nbasesInView - 1;
if(end > seqLength)
end = seqLength;
if(feature_display != null && nbasesInView < feature_display.getMaxVisibleBases())
nbasesInView = feature_display.getMaxVisibleBases();
float pixPerBase = getPixPerBaseByWidth();
boolean changeToStackView = false;
MemoryMXBean memory = ManagementFactory.getMemoryMXBean();
coverageView.init(this, pixPerBase, start, end);
if(isCoverage)
coveragePanel.init(this, pixPerBase, start, end);
if(isSNPplot)
snpPanel.init(this, pixPerBase, start, end);
synchronized (this)
try
{
float heapFractionUsedBefore = (float) ((float) memory.getHeapMemoryUsage().getUsed() /
(float) memory.getHeapMemoryUsage().getMax());
if(readsInView == null)
readsInView = new Vector<SAMRecord>();
else
readsInView.clear();
for(int i=0; i<bamList.size(); i++)
{
if(!hideBamList.contains(i))
readFromBamPicard(start, end, i, pixPerBase);
float heapFractionUsedAfter = (float) ((float) memory.getHeapMemoryUsage().getUsed() /
(float) memory.getHeapMemoryUsage().getMax());
// System.out.println("Heap Max : "+memory.getHeapMemoryUsage().getMax());
// System.out.println("Heap Used : "+memory.getHeapMemoryUsage().getUsed());
// System.out.println("Heap memory used "+heapFractionUsedAfter);
if ((heapFractionUsedAfter - heapFractionUsedBefore) > 0.06
&& !isStackView() && heapFractionUsedAfter > 0.8)
changeToStackView = true;
}
if((!isStackView() && !isStrandStackView()) || isBaseAlignmentView(pixPerBase))
{
Collections.sort(readsInView, new SAMRecordComparator());
}
else if( (isStackView() || isStrandStackView()) &&
bamList.size() > 1)
{
// merge multiple BAM files
Collections.sort(readsInView, new SAMRecordPositionComparator(BamView.this));
}
catch (OutOfMemoryError ome)
JOptionPane.showMessageDialog(this, "Out of Memory");
readsInView.clear();
return;
catch(IOException me)
{
me.printStackTrace();
}
catch(net.sf.samtools.util.RuntimeIOException re)
{
JOptionPane.showMessageDialog(this, re.getMessage());
}
drawCoverage(g2,start, end, pixPerBase);
drawStackView(g2, seqLength, pixPerBase, start, end);
drawPairedStackView(g2, seqLength, pixPerBase, start, end);
else
drawLineView(g2, seqLength, pixPerBase, start, end);
if(isCoverage)
coveragePanel.repaint();
if(isSNPplot)
snpPanel.repaint();
if(changeToStackView)
{
"Note :: Changed to the stack view to save memory.\n"+
"Currently this is using "+
(memory.getHeapMemoryUsage().getUsed()/1000000.f)+" Mb "+
"and the maximum\nmemory limit is "+
(memory.getHeapMemoryUsage().getMax()/1000000.f)+" Mb.",
return (float)mainPanel.getWidth() / (float)nbasesInView;
{
if(feature_display == null)
return seqLength+nbasesInView/3;
/**
* Draw the zoomed-in base view.
* @param g2
* @param seqLength
* @param pixPerBase
* @param start
* @param end
*/
private void drawBaseAlignment(Graphics2D g2,
int seqLength,
float pixPerBase,
final int start,
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int ypos = 0;
end = start + ( mainPanel.getWidth() * ALIGNMENT_PIX_PER_BASE );
if(seqEnd > bases.getLength())
seqEnd = bases.getLength();
bases.getSubSequence(new Range(refSeqStart, seqEnd), Bases.FORWARD).toUpperCase();
ruler.refSeq = refSeq;
}
catch (OutOfRangeException e)
{
e.printStackTrace();
}
}
ruler.repaint();
drawSelectionRange(g2, ALIGNMENT_PIX_PER_BASE, start, end, Color.PINK);
g2.setStroke(new BasicStroke (2.f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND));
Rectangle r = jspView.getViewport().getViewRect();
int nreads = readsInView.size();
ypos += 11;
SAMRecord thisRead = readsInView.get(i);
if (ypos < r.getMaxY() || ypos > r.getMinY())
drawSequence(g2, thisRead, ypos, refSeq, refSeqStart);
drawn[i] = true;
int thisEnd = thisRead.getAlignmentEnd();
if (thisEnd == 0)
thisEnd = thisRead.getAlignmentStart() + thisRead.getReadLength();
for (int j = i + 1; j < nreads; j++)