/**
* First step of overhang assignment - enforce numeric place holders for overhangs, ie no overhang
* redundancy in any step *
*/
private void assignBioBricksOverhangs(
ArrayList<RGraph> optimalGraphs, HashMap<Integer, Vector> stageVectors) {
// Initialize fields that record information to save complexity for future steps
_rootBasicNodeHash = new HashMap<RNode, ArrayList<RNode>>();
HashMap<Integer, RVector> stageRVectors = new HashMap<Integer, RVector>();
for (Integer stage : stageVectors.keySet()) {
RVector vec = ClothoReader.vectorImportClotho(stageVectors.get(stage));
stageRVectors.put(stage, vec);
}
// If the stageVector hash is empty, make a new default vector
if (stageRVectors.size() == 1) {
if (stageRVectors.get(1) == null) {
stageRVectors.put(0, new RVector("EX", "SP", -1, "pSK1A2", null));
}
}
// Loop through each optimal graph and grab the root node to prime for the traversal
for (RGraph graph : optimalGraphs) {
RNode root = graph.getRootNode();
RVector vector = stageRVectors.get(root.getStage() % stageRVectors.size());
RVector rootVector = new RVector("EX", "SP", -1, vector.getName(), null);
root.setVector(rootVector);
root.setLOverhang("EX");
root.setROverhang("SP");
ArrayList<RNode> l0nodes = new ArrayList<RNode>();
_rootBasicNodeHash.put(root, l0nodes);
ArrayList<RNode> neighbors = root.getNeighbors();
assignBioBricksOverhangsHelper(root, neighbors, root, stageRVectors);
}
// Determine which nodes impact which level to form the stageDirectionAssignHash
for (RGraph graph : optimalGraphs) {
RNode root = graph.getRootNode();
ArrayList<String> rootDir = new ArrayList<String>();
ArrayList<String> direction = root.getDirection();
rootDir.addAll(direction);
ArrayList<RNode> l0Nodes = _rootBasicNodeHash.get(root);
// Determine which levels each basic node impacts
for (int i = 0; i < l0Nodes.size(); i++) {
// Determine direction of basic level 0 nodes
RNode l0Node = l0Nodes.get(i);
String l0Direction = rootDir.get(0);
if (l0Node.getComposition().size() == 1) {
ArrayList<String> l0Dir = new ArrayList<String>();
l0Dir.add(l0Direction);
l0Node.setDirection(l0Dir);
}
int size = l0Node.getDirection().size();
rootDir.subList(0, size).clear();
}
}
}
/** Generation of new BioBricks primers for parts * */
public static String[] generateVectorPrimers(
RVector vector,
Collector coll,
Double meltingTemp,
Integer targetLength,
Integer maxPrimerLength,
Integer minPCRLength) {
// initialize primer parameters
String[] oligos = new String[2];
String vectorPrimerPrefix = "gaattcgcggccgcttctagag";
String vectorPrimerSuffix = "tactagtagcggccgctgcag";
Vector currentVector = coll.getVector(vector.getUUID(), true);
String seq = currentVector.getSeq();
String fwdHomology;
String revHomology;
if (seq.equals("")) {
fwdHomology = "[ VECTOR " + currentVector.getName() + " HOMOLOGY REGION ]";
revHomology = "[ VECTOR " + currentVector.getName() + " HOMOLOGY REGION ]";
} else {
fwdHomology =
seq.substring(
0,
Math.min(
seq.length(),
PrimerDesign.getPrimerHomologyLength(
meltingTemp, targetLength, maxPrimerLength - 23, minPCRLength, seq, true)));
revHomology =
PrimerDesign.reverseComplement(
currentVector
.getSeq()
.substring(
Math.max(
0,
seq.length()
- PrimerDesign.getPrimerHomologyLength(
meltingTemp,
targetLength,
maxPrimerLength - 22,
minPCRLength,
PrimerDesign.reverseComplement(seq),
true))));
}
String forwardOligoSequence = vectorPrimerPrefix + fwdHomology;
String reverseOligoSequence = PrimerDesign.reverseComplement(vectorPrimerSuffix) + revHomology;
oligos[0] = forwardOligoSequence;
oligos[1] = reverseOligoSequence;
return oligos;
}
/**
* This helper method executes the loops necessary to enforce overhangs for each graph in
* enforceOverhangRules *
*/
private void assignBioBricksOverhangsHelper(
RNode parent,
ArrayList<RNode> children,
RNode root,
HashMap<Integer, RVector> stageRVectors) {
// Loop through each one of the children to assign rule-instructed overhangs... enumerated
// numbers currently
for (int i = 0; i < children.size(); i++) {
RNode child = children.get(i);
// Give biobricks overhangs
RVector vector = stageRVectors.get(child.getStage() % stageRVectors.size());
RVector newVector = new RVector("EX", "SP", -1, vector.getName(), null);
child.setVector(newVector);
child.setLOverhang("EX");
child.setROverhang("SP");
// Make recursive call
if (child.getStage() > 0) {
ArrayList<RNode> grandChildren = new ArrayList<RNode>();
grandChildren.addAll(child.getNeighbors());
// Remove the current parent from the list
if (grandChildren.contains(parent)) {
grandChildren.remove(parent);
}
assignBioBricksOverhangsHelper(child, grandChildren, root, stageRVectors);
// Or record the level zero parts
} else {
ArrayList<RNode> l0nodes = _rootBasicNodeHash.get(root);
l0nodes.add(child);
_rootBasicNodeHash.put(root, l0nodes);
}
}
}
