1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438
|
#include <iostream>
#include <cstdlib>
#include <string>
#include <cstring>
#include <sstream>
#include <fstream>
#include <map>
#include <set>
#include <vector>
#include <assert.h>
#include "fasta.h"
#include "utils.h"
using namespace std;
const string ERROR_PREFIX = "[REAPR break] ";
string int2string(int number);
struct Breakpoint
{
unsigned long start;
unsigned long end;
short type;
};
struct CmdLineOptions
{
double minTriError;
unsigned long minScaffLength;
unsigned long minMainScaffLength;
bool breakContigs;
unsigned long breakContigsTrim;
bool ignoreContigErrors;
string fastaIn;
string outprefix;
string gff;
};
// deals with command line options: fills the options struct
void parseOptions(int argc, char** argv, CmdLineOptions& ops);
int main(int argc, char** argv)
{
CmdLineOptions options;
parseOptions(argc, argv, options);
Fasta seq;
string line;
map<string, set<pair<unsigned long, unsigned long> > > gapsToBreak;
map<string, set<pair<unsigned long, unsigned long> > > badRegions;
map<string, set<pair<unsigned long, unsigned long> > > contigBreakFlanks;
ifstream inStream;
ofstream outStreamFasta, outStreamBin;
Tabix ti(options.gff);
string fastaOut = options.outprefix + ".broken_assembly.fa";
string binOut = options.outprefix + ".broken_assembly_bin.fa";
vector<pair< string, unsigned long> > refLengths;
string binPrefix = "REAPR_bin.";
// For each chromosome, get an (ordered by coord) list of the gaps to be broken and the
// sections to be replaced by Ns. Some of these regions can intersect.
while (ti.getNextLine(line))
{
vector<string> v;
split(line, '\t', v);
string id = v[0];
// we break when errors are called over a gap, replace with Ns when
// error called in a contig (i.e. not over a gap)
if (v[2].compare("Frag_cov_gap") == 0)
{
// scaffold19_size430204 REAPR Frag_cov_gap 362796 363397 0.00166113 . . Note=Error: Fragment coverage too low over gap 363104-363113;colour=12
vector<string> a,b,c;
split(v.back(), ';', a);
split(a[a.size() - 2], ' ', b);
split(b.back(), ',', c);
for (unsigned long i = 0; i < c.size(); i++)
{
vector<string> d;
split(c[i], '-', d);
unsigned long start = atoi(d[0].c_str()) - 1;
unsigned long end = atoi(d[1].c_str()) - 1;
gapsToBreak[id].insert(make_pair(start, end));
}
}
else if (v[2].compare("FCD_gap") == 0 && atof(v[5].c_str()) >= options.minTriError)
{
// scaffold7_size612284 REAPR FCD_gap 547103 553357 0.781932 . . Note=Error: FCD failure over gap 550388-550397,552547-552686;colour=16
vector<string> a,b,c;
split(v.back(), ';', a);
split(a[a.size() - 2], ' ', b);
split(b.back(), ',', c);
for (unsigned long i = 0; i < c.size(); i++)
{
vector<string> d;
split(c[i], '-', d);
unsigned long start = atoi(d[0].c_str()) - 1;
unsigned long end = atoi(d[1].c_str()) - 1;
gapsToBreak[id].insert(make_pair(start, end));
}
}
else if (!options.ignoreContigErrors &&
(v[2].compare("Frag_cov") == 0 || (v[2].compare("FCD") == 0 && atof(v[5].c_str()) >= options.minTriError)) )
{
// scaffold6_size716595 REAPR Frag_cov 600296 601454 0 . . Note=Error: Fragment coverage too low;color=15
unsigned long start = atoi(v[3].c_str()) - 1;
unsigned long end = atoi(v[4].c_str()) - 1;
if (options.breakContigs)
{
unsigned long middle = 0.5 * (start + end);
gapsToBreak[id].insert(make_pair(middle, middle));
if (options.breakContigsTrim)
{
start = middle >= options.breakContigsTrim ? middle - options.breakContigsTrim : 0;
end = middle + options.breakContigsTrim;
contigBreakFlanks[id].insert(make_pair(start, end));
}
}
else
{
badRegions[id].insert(make_pair(start, end));
}
}
}
// for each chromosome, the replace by Ns errors could intersect. Take the union of them each time two intersect
for (map<string, set<pair<unsigned long, unsigned long> > >::iterator namesIter = badRegions.begin(); namesIter != badRegions.end(); namesIter++)
{
set<pair<unsigned long, unsigned long> > newRegions;
for (set<pair<unsigned long, unsigned long> >::iterator posIter = namesIter->second.begin(); posIter != namesIter->second.end(); posIter++)
{
if (newRegions.size() == 0)
{
newRegions.insert(*posIter);
}
else
{
set<pair<unsigned long, unsigned long> >::iterator lastRegion = newRegions.end();
lastRegion--;
// if last region added intersects with the current region
if (lastRegion->first <= posIter->second && posIter->first <= lastRegion->second)
{
unsigned long start = min(lastRegion->first, posIter->first);
unsigned long end = max(lastRegion->second, posIter->second);
newRegions.erase(lastRegion);
newRegions.insert(make_pair(start, end));
}
else // no overlap
{
newRegions.insert(*posIter);
}
}
}
badRegions[namesIter->first] = newRegions;
}
inStream.open(options.fastaIn.c_str());
if (! inStream.is_open())
{
cerr << ERROR_PREFIX << "Error opening file '" << options.fastaIn << "'" << endl;
return 1;
}
outStreamFasta.open(fastaOut.c_str());
if (! outStreamFasta.is_open())
{
cerr << ERROR_PREFIX << "Error opening file '" << fastaOut << "'" << endl;
return 1;
}
outStreamBin.open(binOut.c_str());
if (! outStreamBin.is_open())
{
cerr << ERROR_PREFIX << "Error opening file '" << binOut << "'" << endl;
return 1;
}
// do the breaking
while (seq.fillFromFile(inStream))
{
map<string, set<pair<unsigned long, unsigned long> > >::iterator gapsToBreakNamesIter = gapsToBreak.find(seq.id);
map<string, set<pair<unsigned long, unsigned long> > >::iterator badRegionsNamesIter = badRegions.find(seq.id);
map<string, set<pair<unsigned long, unsigned long> > >::iterator contigBreakFlanksIter = contigBreakFlanks.find(seq.id);
// replace each region flanking a contig error with Ns. This is only
// relevant if -a and -t were used.
if (contigBreakFlanksIter != contigBreakFlanks.end())
{
for(set<pair<unsigned long, unsigned long> >::iterator p = contigBreakFlanksIter->second.begin(); p != contigBreakFlanksIter->second.end(); p++)
{
unsigned long start = p->first;
unsigned long end = min(p->second, seq.length() - 1);
seq.seq.replace(start, end - start + 1, end - start + 1, 'N');
}
}
// replace the bad regions with Ns, write these sequences out to the "bin" assembly.
// Each of these sequences is broken at any bad gaps flagged. (Sometimes the errors
// over and not over a gap can overlap.)
if (badRegionsNamesIter != badRegions.end())
{
set<pair<unsigned long, unsigned long> >::iterator gapsIter;
if (gapsToBreakNamesIter != gapsToBreak.end())
{
gapsIter = gapsToBreakNamesIter->second.begin();
}
for (set<pair<unsigned long, unsigned long> >::iterator p = badRegionsNamesIter->second.begin(); p != badRegionsNamesIter->second.end(); p++)
{
vector<unsigned long> binRegions;
binRegions.push_back(p->first);
if (gapsToBreakNamesIter != gapsToBreak.end())
{
while (gapsIter != gapsToBreakNamesIter->second.end() && gapsIter->second < p->first)
{
gapsIter++;
}
while (gapsIter != gapsToBreakNamesIter->second.end() && gapsIter->first <= p->second)
{
if (binRegions.size())
{
assert(binRegions.back() < gapsIter->first);
}
binRegions.push_back(max(p->first, gapsIter->first));
binRegions.push_back(min(gapsIter->second, p->second));
gapsIter++;
}
}
binRegions.push_back(p->second);
for (unsigned long i = 0; i < binRegions.size(); i+= 2)
{
Fasta contig = seq.subseq(binRegions[i], binRegions[i+1]);
stringstream ss;
unsigned long startBasesTrimmed, endBasesTrimmed;
contig.trimNs(startBasesTrimmed, endBasesTrimmed);
ss << binRegions[i] + 1 + startBasesTrimmed << '_' << binRegions[i+1] - endBasesTrimmed + 1;
contig.id = binPrefix + contig.id + "_" + ss.str();
if (contig.length() > options.minMainScaffLength)
{
contig.print(outStreamFasta, 60);
}
else if (contig.length() >= options.minScaffLength)
{
contig.print(outStreamBin, 60);
}
}
seq.seq.replace(p->first, p->second - p->first + 1, p->second - p->first + 1, 'N');
}
}
// if there's no breaks to be made at gaps
if (gapsToBreakNamesIter == gapsToBreak.end())
{
unsigned long startBasesTrimmed, endBasesTrimmed;
seq.trimNs(startBasesTrimmed, endBasesTrimmed);
if (startBasesTrimmed || endBasesTrimmed)
{
stringstream ss;
ss << startBasesTrimmed + 1 << '_' << seq.length() - endBasesTrimmed - startBasesTrimmed;
seq.id += "_" + ss.str();
if (seq.length() >= options.minScaffLength)
{
seq.print(outStreamFasta, 60);
}
}
else
{
if (seq.length() >= options.minScaffLength)
{
seq.print(outStreamFasta, 60);
}
}
}
else // gaps to be broken
{
set<pair<unsigned long, unsigned long> > contigCoords;
vector<unsigned long> breakpoints;
breakpoints.push_back(0);
for (set<pair<unsigned long, unsigned long> >::iterator gapsIter = gapsToBreakNamesIter->second.begin(); gapsIter != gapsToBreakNamesIter->second.end(); gapsIter++)
{
if (breakpoints.size())
{
assert(breakpoints.back() <= gapsIter->first);
}
breakpoints.push_back(gapsIter->first == 0 ? 0 : gapsIter->first - 1);
breakpoints.push_back(gapsIter->second + 1);
}
breakpoints.push_back(seq.length() - 1);
for (unsigned long i = 0; i < breakpoints.size(); i+= 2)
{
Fasta contig = seq.subseq(breakpoints[i], breakpoints[i+1]);
stringstream ss;
unsigned long startBasesTrimmed, endBasesTrimmed;
contig.trimNs(startBasesTrimmed, endBasesTrimmed);
ss << breakpoints[i] + 1 + startBasesTrimmed << '_' << breakpoints[i+1] - endBasesTrimmed + 1;
contig.id += "_" + ss.str();
if (contig.length() >= options.minScaffLength)
{
contig.print(outStreamFasta, 60);
}
}
}
}
inStream.close();
outStreamFasta.close();
outStreamBin.close();
return 0;
}
void parseOptions(int argc, char** argv, CmdLineOptions& ops)
{
string usage;
short requiredArgs = 3;
int i;
usage = "\
where 'errors.gff.gz' is the errors gff file made when running score.\n\n\
Options:\n\
-a\n\tAggressive breaking: break contigs at any FCD or low_frag error, as\n\
\topposed to the default of replacing with Ns. Contigs are broken at the\n\
\tmidpoint of each error. Also see option -t. Incompatible with -b\n\
-b\n\tIgnore FCD and low fragment coverage errors that do not contain\n\
\ta gap (the default is to replace these with Ns). incompatible with -a\n\
-e <float>\n\tMinimum FCD error [0]\n\
-l <int>\n\tMinimum sequence length to output [100]\n\
-m <int>\n\tMax sequence length to write to the bin. Sequences longer\n\
\tthan this are written to the main assembly output. This is to stop\n\
\tlong stretches of sequence being lost [999]\n\
-t <int>\n\tWhen -a is used, use this option to specify how many bases\n\
\tare trimmed off the end of each new contig around a break.\n\
\t-t N means that, at an FCD error, a contig is broken at the middle\n\
\tcoordinate of the error, then N bases are\n\
\ttrimmed off each new contig end [0]\n\
";
if (argc == 2 && strcmp(argv[1], "--wrapperhelp") == 0)
{
usage = "[options] <assembly.fa> <errors.gff.gz> <outfiles prefix>\n\n" + usage;
cerr << usage << endl;
exit(1);
}
else if (argc < requiredArgs)
{
usage = "[options] <assembly.fa> <errors.gff.gz> <outfiles prefix>\n\n" + usage;
cerr << "usage: task_break " << usage;
exit(1);
}
ops.minTriError = 0;
ops.minScaffLength = 100;
ops.minMainScaffLength = 999;
ops.breakContigs = false;
ops.ignoreContigErrors = false;
ops.breakContigsTrim = 0;
for (i = 1; i < argc - requiredArgs; i++)
{
if (strcmp(argv[i], "-a") == 0)
{
ops.breakContigs = true;
continue;
}
if (strcmp(argv[i], "-b") == 0)
{
ops.ignoreContigErrors = true;
continue;
}
if (strcmp(argv[i], "-e") == 0)
{
ops.minTriError = atof(argv[i+1]);
}
else if (strcmp(argv[i], "-l") == 0)
{
ops.minScaffLength = atoi(argv[i+1]);
}
else if (strcmp(argv[i], "-m") == 0)
{
ops.minMainScaffLength = atoi(argv[i+1]);
}
else if (strcmp(argv[i], "-t") == 0)
{
ops.breakContigsTrim = atoi(argv[i+1]);
}
else
{
cerr << ERROR_PREFIX << "Error! Switch not recognised: " << argv[i] << endl;
exit(1);
}
i++;
}
if (argc - i != requiredArgs || argv[i+1][0] == '-')
{
cerr << usage;
exit(1);
}
if (ops.ignoreContigErrors && ops.breakContigs)
{
cerr << ERROR_PREFIX << "Options -a and -b are incompatible. Cannot continue" << endl;
exit(1);
}
if (ops.breakContigsTrim && !ops.breakContigs)
{
cerr << ERROR_PREFIX << "Warning: ignoring -t " << ops.breakContigsTrim << " because -a was not used" << endl;
}
ops.fastaIn = argv[i];
ops.gff = argv[i+1];
ops.outprefix = argv[i+2];
}
string int2string(int number)
{
stringstream ss;
ss << number;
return ss.str();
}
|