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root/gclib/gclib/GAlnExtend.cpp
Revision: 176
Committed: Thu Feb 16 03:47:16 2012 UTC (7 years, 5 months ago) by gpertea
File size: 40306 byte(s)
Log Message:
fqtrim reworked

Line User Rev File contents
1 gpertea 93 #include "GAlnExtend.h"
2    
3     //greedy gapped alignment extension
4     //(mostly lifted from NCBI's megablast gapped extension code)
5    
6     int GXMemPool::kMinSpace = 1000000;
7    
8     #ifdef GDEBUG
9     char buf[6]={0x1B,'[', 'n','m','m','\0'};
10    
11     void color_fg(int c,FILE* f) {
12     if (f!=stderr && f!=stdout) return;
13     sprintf((char *)(&buf[2]),"%dm",c+30);
14     fwrite(buf,1,strlen(buf), f);
15     }
16    
17     void color_bg(int c, FILE* f) {
18     if (f!=stderr && f!=stdout) return;
19     sprintf((char *)(&buf[2]),"%dm",c+40);
20     fwrite(buf,1,strlen(buf),f);
21     };
22    
23     void color_resetfg(FILE* f) {
24     if (f!=stderr && f!=stdout) return;
25     sprintf((char *)(&buf[2]),"39m");
26     fwrite(buf,1,strlen(buf), f);
27     };
28    
29     void color_resetbg(FILE* f) {
30     if (f!=stderr && f!=stdout) return;
31     sprintf((char *)(&buf[2]),"49m");
32     fwrite(buf,1,strlen(buf), f);
33     }
34    
35     void color_reset(FILE* f) {
36     if (f!=stderr && f!=stdout) return;
37     sprintf((char *)(&buf[2]),"0m");
38     fwrite(buf,1,strlen(buf), f);
39     };
40    
41     void color_normal(FILE* f) {
42     if (f!=stderr && f!=stdout) return;
43     sprintf((char *)(&buf[2]),"22m");
44     fwrite(buf,1,strlen(buf), f);
45     };
46    
47     #endif
48    
49    
50     char xgapcodes[4]={'S','I', 'D', 'X'};
51    
52     int get_last(int **flast_d, int d, int diag, int *row1) {
53     if (flast_d[d-1][diag-1] > GMAX(flast_d[d-1][diag], flast_d[d-1][diag+1])) {
54     *row1 = flast_d[d-1][diag-1];
55     return diag-1;
56 gpertea 171 }
57 gpertea 93 if (flast_d[d-1][diag] > flast_d[d-1][diag+1]) {
58     *row1 = flast_d[d-1][diag];
59     return diag;
60     }
61     *row1 = flast_d[d-1][diag+1];
62     return diag+1;
63     }
64    
65     void GapXEditScript::print() { //debug
66     GapXEditScript* p=this;
67     do {
68 gpertea 171 GMessage("%d%c ",p->num, xgapcodes[p->op_type]);
69     } while ((p=p->next)!=NULL);
70 gpertea 93 GMessage("\n");
71     }
72    
73    
74     int BLAST_Gcd(int a, int b) {
75     int c;
76    
77     b = abs(b);
78     if (b > a)
79     c=a, a=b, b=c;
80    
81     while (b != 0) {
82     c = a%b;
83     a = b;
84     b = c;
85     }
86     return a;
87     }
88    
89     int BLAST_Gdb3(int* a, int* b, int* c) {
90     int g;
91     if (*b == 0)
92     g = BLAST_Gcd(*a, *c);
93     else
94     g = BLAST_Gcd(*a, BLAST_Gcd(*b, *c));
95     if (g > 1) {
96 gpertea 173 *a /= g;
97 gpertea 93 *b /= g;
98     *c /= g;
99     }
100     return g;
101     }
102    
103    
104 gpertea 171 uint16 get6mer(char* p) {
105     uint16 r=gdna2bit(p,3);
106 gpertea 173 r <<= 6;
107     r |= gdna2bit(p,3);
108 gpertea 171 return r;
109     }
110 gpertea 93
111 gpertea 173
112     void table6mers(const char* s, int slen, GVec<uint16>* amers[]) {
113 gpertea 174 for (uint16 i=0; i <= slen-6; i++) {
114 gpertea 173 char* p = (char*)(s+i);
115     uint16 v=get6mer(p);
116     if (amers[v]==NULL) {
117     amers[v]=new GVec<uint16>(1);
118     }
119     amers[v]->Add(i);
120     }
121     }
122    
123     GVec<uint16>* match6mer(char* start, GVec<uint16>* amers[]) {
124     //careful: this is broken if start+5 falls beyond the end of the string!
125     uint16 r=get6mer(start);
126     return amers[r];
127     }
128    
129 gpertea 93 //signal that a diagonal is invalid
130     static const int kInvalidOffset = -2;
131    
132     int s_FindFirstMismatch(const char *seq1, int len1,
133     const char *seq2, int len2,
134     int seq1_index, int seq2_index,
135     //bool &fence_hit,
136     bool reverse) {
137     int start_index = seq1_index;
138     /* Sentry detection here should be relatively inexpensive: The
139     sentry value cannot appear in the query, so detection only
140     needs to be done at exit from the subject-query matching loop.
141     For uncompressed sequences, ambiguities in the query (i.e. seq1)
142     always count as mismatches */
143     if (reverse) {
144     while (seq1_index < len1 && seq2_index < len2 &&
145     //seq1[len1-1 - seq1_index] < 4 &&
146     seq1[len1-1 - seq1_index] == seq2[len2-1 - seq2_index]) {
147     ++seq1_index;
148     ++seq2_index;
149     }
150     //if (seq2_index < len2 && seq2[len2-1-seq2_index] == FENCE_SENTRY) {
151     //if len2-1-seq2_index<=0) {
152     // fence_hit = true;
153     // }
154     }
155     else { //forward lookup
156 gpertea 171 while (seq1_index < len1 && seq2_index < len2 &&
157 gpertea 93 //seq1[seq1_index] < 4 &&
158     seq1[seq1_index] == seq2[seq2_index]) {
159     ++seq1_index;
160     ++seq2_index;
161     }
162     //if (seq2_index < len2 && seq2[seq2_index] == FENCE_SENTRY) {
163     //if (seq2_index==len2) {
164     // fence_hit = true;
165     //}
166     }
167     return seq1_index - start_index;
168     }
169    
170    
171    
172     /** During the traceback for a non-affine greedy alignment,
173     compute the diagonal that will result from the next
174     traceback operation
175    
176     @param last_seq2_off Array of offsets into the second sequence;
177     last_seq2_off[d][k] gives the largest offset into
178     the second sequence that lies on diagonal k and
179     has distance d [in]
180     @param d Starting distance [in]
181     @param diag Index of diagonal that produced the starting distance [in]
182     @param seq2_index The offset into the second sequence after the traceback
183     operation has completed [out]
184     @return The diagonal resulting from the next traceback operation
185     being applied
186     */
187     int s_GetNextNonAffineTback(int **last_seq2_off, int d,
188     int diag, int *seq2_index) {
189     // choose the traceback operation that results in the
190     // largest seq2 offset at this point, then compute the
191     // new diagonal that is implied by the operation
192     if (last_seq2_off[d-1][diag-1] >
193     GMAX(last_seq2_off[d-1][diag], last_seq2_off[d-1][diag+1])) {
194     *seq2_index = last_seq2_off[d-1][diag-1];
195     return diag - 1; // gap in seq2
196     }
197     if (last_seq2_off[d-1][diag] > last_seq2_off[d-1][diag+1]) {
198     *seq2_index = last_seq2_off[d-1][diag];
199     return diag; // match
200     }
201     *seq2_index = last_seq2_off[d-1][diag+1];
202     return diag + 1; // gap in seq1
203     }
204    
205    
206     int GXGreedyExtend(const char* seq1, int len1,
207     const char* seq2, int len2,
208     bool reverse, int xdrop_threshold,
209     int match_cost, int mismatch_cost,
210     int& seq1_align_len, int& seq2_align_len,
211 gpertea 101 CGreedyAlignData& aux_data,
212 gpertea 93 GXEditScript *edit_block) {
213     //GapPrelimEditBlock *edit_block,
214     //bool& fence_hit, SGreedySeed *seed) {
215     int seq1_index;
216     int seq2_index;
217     int index;
218     int d;
219     int k;
220     int diag_lower, diag_upper;
221     int max_dist;
222     int diag_origin;
223     int best_dist;
224     int best_diag;
225     int** last_seq2_off;
226     int* max_score;
227     int xdrop_offset;
228     int longest_match_run;
229     bool end1_reached, end2_reached;
230     GXMemPool* mem_pool;
231 gpertea 171
232 gpertea 93 /* ordinary dynamic programming alignment, for each offset
233     in seq1, walks through offsets in seq2 until an X-dropoff
234 gpertea 171 test fails, saving the best score encountered along
235 gpertea 93 the way. Instead of score, this code tracks the 'distance'
236     (number of mismatches plus number of gaps) between seq1
237     and seq2. Instead of walking through sequence offsets, it
238     walks through diagonals that can achieve a given distance.
239 gpertea 171
240 gpertea 93 Note that in what follows, the numbering of diagonals implies
241 gpertea 171 a dot matrix where increasing seq1 offsets go to the right on
242 gpertea 93 the x axis, and increasing seq2 offsets go up the y axis.
243     The gapped alignment thus proceeds up and to the right in
244     the graph, and diagonals are numbered increasing to the right */
245    
246     best_dist = 0;
247     best_diag = 0;
248    
249     /* set the number of distinct distances the algorithm will
250 gpertea 171 examine in the search for an optimal alignment. The
251     heuristic worst-case running time of the algorithm is
252 gpertea 93 O(max_dist**2 + (len1+len2)); for sequences which are
253     very similar, the average running time will be sig-
254     nificantly better than this */
255    
256     max_dist = GMIN(GREEDY_MAX_COST,
257 gpertea 109 (len2/GREEDY_MAX_COST_FRACTION + 1));
258 gpertea 93
259     /* the main loop assumes that the index of all diagonals is
260 gpertea 171 biased to lie in the middle of allocated bookkeeping
261 gpertea 93 structures */
262    
263     diag_origin = max_dist + 2;
264    
265     // last_seq2_off[d][k] is the largest offset into seq2 that
266     // lies on diagonal k and has distance d
267    
268     last_seq2_off = aux_data.last_seq2_off;
269    
270     /* Instead of tracking the best alignment score and using
271     xdrop_theshold directly, track the best score for each
272     unique distance and use the best score for some previously
273     computed distance to implement the X-dropoff test.
274    
275     xdrop_offset gives the distance backwards in the score
276     array to look */
277    
278 gpertea 171 xdrop_offset = (xdrop_threshold + match_cost / 2) /
279 gpertea 93 (match_cost + mismatch_cost) + 1;
280 gpertea 171
281 gpertea 93 // find the offset of the first mismatch between seq1 and seq2
282    
283     index = s_FindFirstMismatch(seq1, len1, seq2, len2, 0, 0, reverse);
284     // fence_hit, reverse, rem);
285    
286     // update the extents of the alignment, and bail out
287     // early if no further work is needed
288    
289     seq1_align_len = index;
290     seq2_align_len = index;
291     seq1_index = index;
292     /*
293     seed->start_q = 0;
294     seed->start_s = 0;
295     seed->match_length = index;
296     */
297     longest_match_run = index;
298    
299     if (index == len1 || index == len2) {
300     /* Return the number of differences, which is zero here */
301     if (edit_block != NULL)
302     //GapPrelimEditBlockAdd(edit_block, eGapAlignSub, index);
303     edit_block->opRep(index);
304 gpertea 171 return 0;
305 gpertea 93 }
306    
307     // set up the memory pool
308     mem_pool = aux_data.space;
309     if (edit_block == NULL) {
310     mem_pool = NULL;
311     }
312     else if (mem_pool == NULL) {
313     aux_data.space = mem_pool = new GXMemPool();
314 gpertea 171 }
315 gpertea 93 else {
316     mem_pool->refresh();
317     }
318 gpertea 171
319 gpertea 93 /* set up the array of per-distance maximum scores. There
320     are max_diags + xdrop_offset distances to track, the first
321     xdrop_offset of which are 0 */
322    
323     max_score = aux_data.max_score + xdrop_offset;
324     for (index = 0; index < xdrop_offset; index++)
325     aux_data.max_score[index] = 0;
326 gpertea 171
327 gpertea 93 // fill in the initial offsets of the distance matrix
328    
329     last_seq2_off[0][diag_origin] = seq1_index;
330     max_score[0] = seq1_index * match_cost;
331     diag_lower = diag_origin - 1;
332     diag_upper = diag_origin + 1;
333     end1_reached = end2_reached = false;
334    
335     // for each distance
336     for (d = 1; d <= max_dist; d++) {
337     int xdrop_score;
338     int curr_score;
339     int curr_extent = 0;
340     int curr_seq2_index = 0;
341     int curr_diag = 0;
342     int tmp_diag_lower = diag_lower;
343     int tmp_diag_upper = diag_upper;
344    
345     // Assign impossible seq2 offsets to any diagonals that
346     // are not in the range (diag_lower,diag_upper).
347     // These will serve as sentinel values for the inner loop
348     last_seq2_off[d - 1][diag_lower-1] = kInvalidOffset;
349     last_seq2_off[d - 1][diag_lower] = kInvalidOffset;
350     last_seq2_off[d - 1][diag_upper] = kInvalidOffset;
351     last_seq2_off[d - 1][diag_upper+1] = kInvalidOffset;
352    
353     // compute the score for distance d corresponding to the X-dropoff criterion
354    
355 gpertea 171 xdrop_score = max_score[d - xdrop_offset] +
356 gpertea 93 (match_cost + mismatch_cost) * d - xdrop_threshold;
357 gpertea 171 xdrop_score = (int)ceil((double)xdrop_score / (match_cost / 2));
358 gpertea 93
359     // for each diagonal of interest
360     for (k = tmp_diag_lower; k <= tmp_diag_upper; k++) {
361     /* find the largest offset into seq2 that increases
362     the distance from d-1 to d (i.e. keeps the alignment
363 gpertea 171 from getting worse for as long as possible), then
364 gpertea 93 choose the offset into seq1 that will keep the
365 gpertea 171 resulting diagonal fixed at k
366    
367 gpertea 93 Note that this requires kInvalidOffset+1 to be smaller
368     than any valid offset into seq2, i.e. to be negative */
369    
370     seq2_index = GMAX(last_seq2_off[d - 1][k + 1],
371     last_seq2_off[d - 1][k ]) + 1;
372     seq2_index = GMAX(seq2_index, last_seq2_off[d - 1][k - 1]);
373     seq1_index = seq2_index + k - diag_origin;
374    
375     if (seq2_index < 0 || seq1_index + seq2_index < xdrop_score) {
376    
377     // if no valid diagonal can reach distance d, or the
378     // X-dropoff test fails, narrow the range of diagonals
379     // to test and skip to the next diagonal
380     if (k == diag_lower)
381     diag_lower++;
382     else
383     last_seq2_off[d][k] = kInvalidOffset;
384     continue;
385     }
386     diag_upper = k;
387 gpertea 171
388     /* slide down diagonal k until a mismatch
389 gpertea 93 occurs. As long as only matches are encountered,
390     the current distance d will not change */
391    
392     index = s_FindFirstMismatch(seq1, len1, seq2, len2,
393     seq1_index, seq2_index, reverse);
394     //fence_hit, reverse, rem);
395     if (index > longest_match_run) {
396     //seed->start_q = seq1_index;
397     //seed->start_s = seq2_index;
398     //seed->match_length = index;
399     longest_match_run = index;
400     }
401     seq1_index += index;
402     seq2_index += index;
403    
404     // set the new largest seq2 offset that achieves
405     // distance d on diagonal k
406    
407     last_seq2_off[d][k] = seq2_index;
408    
409     // since all values of k are constrained to have the
410     // same distance d, the value of k which maximizes the
411     // alignment score is the one that covers the most of seq1 and seq2
412     if (seq1_index + seq2_index > curr_extent) {
413     curr_extent = seq1_index + seq2_index;
414     curr_seq2_index = seq2_index;
415     curr_diag = k;
416     }
417    
418     /* clamp the bounds on diagonals to avoid walking off
419     either sequence. Because the bounds increase by at
420     most one for each distance, diag_lower and diag_upper
421     can each be of size at most max_diags+2 */
422    
423     if (seq2_index == len2) {
424 gpertea 171 diag_lower = k + 1;
425 gpertea 93 end2_reached = true;
426     }
427     if (seq1_index == len1) {
428 gpertea 171 diag_upper = k - 1;
429 gpertea 93 end1_reached = true;
430     }
431     } // end loop over diagonals
432    
433     // compute the maximum score possible for distance d
434 gpertea 171 curr_score = curr_extent * (match_cost / 2) -
435 gpertea 93 d * (match_cost + mismatch_cost);
436     // if this is the best score seen so far, update the
437     // statistics of the best alignment
438     if (curr_score > max_score[d - 1]) {
439     max_score[d] = curr_score;
440     best_dist = d;
441     best_diag = curr_diag;
442     seq2_align_len = curr_seq2_index;
443     seq1_align_len = curr_seq2_index + best_diag - diag_origin;
444 gpertea 171 }
445 gpertea 93 else {
446     max_score[d] = max_score[d - 1];
447     }
448    
449     // alignment has finished if the lower and upper bounds
450     // on diagonals to check have converged to each other
451    
452     if (diag_lower > diag_upper)
453     break;
454    
455 gpertea 171 /* set up for the next distance to examine. Because the
456     bounds increase by at most one for each distance,
457     diag_lower and diag_upper can each be of size at
458 gpertea 93 most max_diags+2 */
459    
460     if (!end2_reached)
461 gpertea 171 diag_lower--;
462 gpertea 93 if (!end1_reached)
463     diag_upper++;
464    
465     if (edit_block == NULL) {
466     // if no traceback is specified, the next row of
467     // last_seq2_off can reuse previously allocated memory
468     //TODO FIXME The following assumes two arrays of
469     // at least max_dist+4 int's have already been allocated
470     last_seq2_off[d + 1] = last_seq2_off[d - 1];
471     }
472     else {
473     // traceback requires all rows of last_seq2_off to be saved,
474     // so a new row must be allocated
475     last_seq2_off[d + 1] = (int*)mem_pool->getByteSpace((diag_upper - diag_lower + 7)*sizeof(int));
476     // move the origin for this row backwards
477     //TODO FIXME: dubious pointer arithmetic ?!
478     //last_seq2_off[d + 1] = last_seq2_off[d + 1] - diag_lower + 2;
479     }
480     } // end loop over distinct distances
481    
482 gpertea 171
483 gpertea 93 if (edit_block == NULL)
484     return best_dist;
485    
486     //---- perform traceback
487 gpertea 171 d = best_dist;
488 gpertea 93 seq1_index = seq1_align_len;
489     seq2_index = seq2_align_len;
490     // for all positive distances
491    
492     //if (fence_hit && *fence_hit)
493     // goto done;
494     if (index==len1 || index==len2) d=0;
495     while (d > 0) {
496     int new_diag;
497     int new_seq2_index;
498    
499     /* retrieve the value of the diagonal after the next
500     traceback operation. best_diag starts off with the
501     value computed during the alignment process */
502    
503 gpertea 171 new_diag = s_GetNextNonAffineTback(last_seq2_off, d,
504 gpertea 93 best_diag, &new_seq2_index);
505    
506     if (new_diag == best_diag) {
507     // same diagonal: issue a group of substitutions
508     if (seq2_index - new_seq2_index > 0) {
509     edit_block->opRep(seq2_index - new_seq2_index);
510     }
511 gpertea 171 }
512 gpertea 93 else if (new_diag < best_diag) {
513     // smaller diagonal: issue a group of substitutions
514     // and then a gap in seq2 */
515     if (seq2_index - new_seq2_index > 0) {
516     edit_block->opRep(seq2_index - new_seq2_index);
517     }
518     //GapPrelimEditBlockAdd(edit_block, eGapAlignIns, 1);
519     edit_block->opIns(1);
520 gpertea 171 }
521 gpertea 93 else {
522     // larger diagonal: issue a group of substitutions
523     // and then a gap in seq1
524     if (seq2_index - new_seq2_index - 1 > 0) {
525     edit_block->opRep(seq2_index - new_seq2_index - 1);
526     }
527     edit_block->opDel(1);
528     }
529 gpertea 171 d--;
530     best_diag = new_diag;
531     seq2_index = new_seq2_index;
532 gpertea 93 }
533     //done:
534     // handle the final group of substitutions back to distance zero,
535     // i.e. back to offset zero of seq1 and seq2
536     //GapPrelimEditBlockAdd(edit_block, eGapAlignSub,
537     // last_seq2_off[0][diag_origin]);
538     edit_block->opRep(last_seq2_off[0][diag_origin]);
539     if (!reverse)
540     edit_block->reverse();
541     return best_dist;
542     }
543    
544     void printEditScript(GXEditScript* ed_script) {
545     uint i;
546     if (ed_script==NULL || ed_script->opnum == 0)
547     return;
548     for (i=0; i<ed_script->opnum; i++) {
549     int num=((ed_script->ops[i]) >> 2);
550     unsigned char op_type = 3 - ( ed_script->ops[i] & gxEDIT_OP_MASK );
551     if (op_type == 3)
552     GError("Error: printEditScript encountered op_type 3 ?!\n");
553 gpertea 171 GMessage("%d%c ", num, xgapcodes[op_type]);
554 gpertea 93 }
555     GMessage("\n");
556     }
557    
558     GXAlnInfo* GreedyAlign(const char* q_seq, int q_alnstart, const char* s_seq, int s_alnstart,
559     bool editscript, int reward, int penalty, int xdrop) {
560     int q_max=strlen(q_seq); //query
561     int s_max=strlen(s_seq); //subj
562     return GreedyAlignRegion(q_seq, q_alnstart, q_max,
563 gpertea 101 s_seq, s_alnstart, s_max, reward, penalty, xdrop, NULL, NULL, editscript);
564 gpertea 93 }
565    
566     struct GXSeedTable {
567     int a_num, b_num;
568     int a_cap, b_cap;
569     char* xc;
570     GXSeedTable(int a=12, int b=255) {
571     a_cap=0;
572     b_cap=0;
573     a_num=0;
574     b_num=0;
575     xc=NULL;
576     init(a,b);
577     }
578     ~GXSeedTable() {
579     GFREE(xc);
580     }
581     void init(int a, int b) {
582     a_num=a;
583     b_num=b;
584     bool resize=false;
585     if (b_num>b_cap) { resize=true; b_cap=b_num;}
586     if (a_num>a_cap) { resize=true; a_cap=a_num;}
587     if (resize) {
588     GFREE(xc);
589     GCALLOC(xc, (a_num*b_num));
590     }
591     else {
592     //just clear up to a_max, b_max
593     memset((void*)xc, 0, (a_num*b_num));
594     }
595     }
596     char& x(int ax, int by) {
597     return xc[by*a_num+ax];
598     }
599    
600     };
601    
602 gpertea 99 const int a_m_score=2; //match score
603     const int a_mis_score=-3; //mismatch
604     const int a_dropoff_score=7;
605 gpertea 101 const int a_min_score=12; //at least 6 bases full match
606 gpertea 93
607 gpertea 99 // ------------------ adapter matching - simple k-mer seed & extend, no indels for now
608     //when a k-mer match is found, simply try to extend the alignment using a drop-off scheme
609     //check minimum score and
610     //for 3' adapter trimming:
611     // require that the right end of the alignment for either the adaptor OR the read must be
612     // < 3 distance from its right end
613     // for 5' adapter trimming:
614     // require that the left end of the alignment for either the adaptor OR the read must
615     // be at coordinate < 3 from start
616    
617     bool extendUngapped(const char* a, int alen, int ai,
618     const char* b, int blen, int bi, int& mlen, int& l5, int& l3, bool end5=false) {
619     //so the alignment starts at ai in a, bi in b, with a perfect match of length mlen
620     //if (debug) {
621     // GMessage(">> in %s\n\textending hit: %s at position %d\n", a, (dbg.substr(bi, mlen)).chars(), ai);
622     // }
623     int a_l=ai; //alignment coordinates on a
624     int a_r=ai+mlen-1;
625     int b_l=bi; //alignment coordinates on b
626     int b_r=bi+mlen-1;
627     int ai_maxscore=ai;
628     int bi_maxscore=bi;
629     int score=mlen*a_m_score;
630     int maxscore=score;
631     int mism5score=a_mis_score;
632     if (end5 && ai<(alen>>1)) mism5score-=2; // increase penalty for mismatches at 5' end
633     //try to extend to the left first, if possible
634     while (ai>0 && bi>0) {
635     ai--;
636     bi--;
637     score+= (a[ai]==b[bi])? a_m_score : mism5score;
638     if (score>maxscore) {
639     ai_maxscore=ai;
640     bi_maxscore=bi;
641     maxscore=score;
642     }
643     else if (maxscore-score>a_dropoff_score) break;
644     }
645     a_l=ai_maxscore;
646     b_l=bi_maxscore;
647     //now extend to the right
648     ai_maxscore=a_r;
649     bi_maxscore=b_r;
650     ai=a_r;
651     bi=b_r;
652     score=maxscore;
653     //sometimes there are extra As at the end of the read, ignore those
654     if (a[alen-2]=='A' && a[alen-1]=='A') {
655     alen-=2;
656     while (a[alen-1]=='A' && alen>ai) alen--;
657     }
658     while (ai<alen-1 && bi<blen-1) {
659     ai++;
660     bi++;
661     //score+= (a[ai]==b[bi])? a_m_score : a_mis_score;
662     if (a[ai]==b[bi]) { //match
663     score+=a_m_score;
664     if (ai>=alen-2) {
665     score+=a_m_score-(alen-ai-1);
666     }
667     }
668     else { //mismatch
669     score+=a_mis_score;
670     }
671     if (score>maxscore) {
672     ai_maxscore=ai;
673     bi_maxscore=bi;
674     maxscore=score;
675     }
676     else if (maxscore-score>a_dropoff_score) break;
677     }
678     a_r=ai_maxscore;
679     b_r=bi_maxscore;
680     int a_ovh3=alen-a_r-1;
681     int b_ovh3=blen-b_r-1;
682     int mmovh3=(a_ovh3<b_ovh3)? a_ovh3 : b_ovh3;
683     int mmovh5=(a_l<b_l)? a_l : b_l;
684     if (maxscore>=a_min_score && mmovh3<2 && mmovh5<2) {
685     if (a_l<a_ovh3) {
686     //adapter closer to the left end (typical for 5' adapter)
687     l5=a_r+1;
688     l3=alen-1;
689     }
690     else {
691     //adapter matching at the right end (typical for 3' adapter)
692     l5=0;
693     l3=a_l-1;
694     }
695     return true;
696     }
697     //do not trim:
698     l5=0;
699     l3=alen-1;
700     return false;
701     }
702    
703    
704 gpertea 176 GXBandSet* collectSeeds_R(GList<GXSeed>& seeds, GXSeqData& sd) {
705     int bimin=GMAX(0,(sd.blen-sd.alen-6));
706     GXSeedTable gx(sd.alen, sd.blen);
707     GXBandSet* diagstrips=new GXBandSet(sd.alen, sd.blen); //set of overlapping 3-diagonal strips
708     //for (int bi=0;bi<=b_len-6;bi++) {
709     for (int bi=sd.blen-6;bi>=0;bi--) {
710 gpertea 174 //for each 6-mer of seqb
711 gpertea 176 uint16 bv = get6mer((char*)&(sd.bseq[bi]));
712     GVec<uint16>* alocs = sd.amers[bv];
713 gpertea 174 if (alocs==NULL) continue;
714     //extend each hit
715     for (int h=0;h<alocs->Count();h++) {
716     int ai=alocs->Get(h);
717     //word match
718     if (gx.x(ai,bi))
719     //already have a previous seed covering this region of this diagonal
720     continue;
721     for (int i=0;i<6;i++)
722     gx.x(ai+i,bi+i)=1;
723 gpertea 176 //see if we can extend to the left
724     int aix=ai-1;
725     int bix=bi-1;
726 gpertea 174 int len=6;
727 gpertea 176 while (bix>=0 && aix>=0 && sd.aseq[aix]==sd.bseq[bix]) {
728 gpertea 174 gx.x(aix,bix)=1;
729 gpertea 176 ai=aix;
730     bi=bix;
731     aix--;bix--;
732 gpertea 174 len++;
733     }
734 gpertea 176 if (len>sd.amlen) {
735     //heuristics: likely the best we can get
736 gpertea 174 //quick match shortcut
737     diagstrips->qmatch=new GXSeed(ai,bi,len);
738     return diagstrips;
739 gpertea 176 }
740 gpertea 174 if (bi<bimin && len<9) continue; //skip middle seeds that are not high scoring enough
741     GXSeed* newseed=new GXSeed(ai,bi,len);
742     seeds.Add(newseed);
743     diagstrips->addSeed(newseed);//add it to all 3 adjacent diagonals
744     //special last resort terminal match to be used if no better alignment is there
745 gpertea 176 if (ai<2 && bi+len>sd.blen-2 &&
746 gpertea 174 (!diagstrips->tmatch || diagstrips->tmatch->len<len)) diagstrips->tmatch=newseed;
747     }
748     }
749 gpertea 93 for (int i=0;i<diagstrips->Count();i++) {
750     diagstrips->Get(i)->finalize(); //adjust scores due to overlaps or gaps between seeds
751     }
752     diagstrips->setSorted(true); //sort by score
753     return diagstrips;
754     }
755    
756 gpertea 176 GXBandSet* collectSeeds_L(GList<GXSeed>& seeds, GXSeqData& sd) {
757 gpertea 93 //overlap of left (5') end of seqb
758     //hash the last 12 bases of seqa:
759 gpertea 176 int bimax=GMIN((sd.alen+2), (sd.blen-6));
760 gpertea 93 //gx.init(a_maxlen, b_maxlen);
761 gpertea 176 GXSeedTable gx(sd.alen, sd.blen);
762     GXBandSet* diagstrips=new GXBandSet(sd.alen, sd.blen); //set of overlapping 3-diagonal strips
763     for (int bi=0;bi<=sd.blen-6;bi++) {
764 gpertea 174 //for each 6-mer of seqb
765 gpertea 176 uint16 bv = get6mer((char*) & (sd.bseq[bi]));
766     GVec<uint16>* alocs = sd.amers[bv];
767 gpertea 174 if (alocs==NULL) continue;
768     //extend each hit
769     for (int h=0;h<alocs->Count();h++) {
770     int ai=alocs->Get(h);
771     //word match
772     if (gx.x(ai,bi))
773     //already have a previous seed covering this region of this diagonal
774     continue;
775     for (int i=0;i<6;i++)
776     gx.x(ai+i,bi+i)=1;
777     //see if we can extend to the right
778     int aix=ai+6;
779     int bix=bi+6;
780     int len=6;
781 gpertea 176 while (bix<sd.blen && aix<sd.alen && sd.aseq[aix]==sd.bseq[bix]) {
782 gpertea 174 gx.x(aix,bix)=1;
783     aix++;bix++;
784     len++;
785     }
786 gpertea 176 if (len>sd.amlen) {
787 gpertea 174 //heuristics: very likely the best we can get
788     //quick match shortcut
789     diagstrips->qmatch=new GXSeed(ai,bi,len);
790     return diagstrips;
791 gpertea 176 }
792 gpertea 174 if (bi>bimax && len<9) continue; //skip middle seeds that are not high scoring enough
793     GXSeed* newseed=new GXSeed(ai,bi,len);
794     seeds.Add(newseed);
795     diagstrips->addSeed(newseed);//add it to all 3 adjacent diagonals
796     //special last resort terminal match to be used if no better alignment is there
797 gpertea 176 if (bi<2 && ai+len>=sd.alen-1 &&
798 gpertea 174 (!diagstrips->tmatch || diagstrips->tmatch->len<len))
799     diagstrips->tmatch=newseed;
800     }
801     } //for each 6-mer of the read
802 gpertea 93 for (int i=0;i<diagstrips->Count();i++) {
803     diagstrips->Get(i)->finalize(); //adjust scores due to overlaps or gaps between seeds
804     }
805     diagstrips->setSorted(true); //sort by score
806     return diagstrips;
807     }
808    
809     int cmpSeedScore(const pointer p1, const pointer p2) {
810     //return (((GXSeed*)s2)->len-((GXSeed*)s1)->len);
811     GXSeed* s1=(GXSeed*)p1;
812     GXSeed* s2=(GXSeed*)p2;
813     if (s1->len==s2->len) {
814     return (s1->b_ofs-s2->b_ofs);
815     }
816     else return (s2->len-s1->len);
817     }
818    
819 gpertea 174 int cmpSeedScore_R(const pointer p1, const pointer p2) {
820     //return (((GXSeed*)s2)->len-((GXSeed*)s1)->len);
821     GXSeed* s1=(GXSeed*)p1;
822     GXSeed* s2=(GXSeed*)p2;
823     if (s1->len==s2->len) {
824     return (s2->b_ofs-s1->b_ofs);
825     }
826     else return (s2->len-s1->len);
827     }
828    
829    
830 gpertea 93 int cmpSeedDiag(const pointer p1, const pointer p2) {
831     GXSeed* s1=(GXSeed*)p1;
832     GXSeed* s2=(GXSeed*)p2;
833     return ((s1->b_ofs-s1->a_ofs)-(s2->b_ofs-s2->a_ofs));
834     }
835    
836 gpertea 101
837     int cmpDiagBands_R(const pointer p1, const pointer p2) {
838     //return (((GXSeed*)s2)->len-((GXSeed*)s1)->len);
839     GXBand* b1=(GXBand*)p1;
840     GXBand* b2=(GXBand*)p2;
841     if (b1->score==b2->score) {
842     return (b2->w_min_b-b1->w_min_b);
843     }
844     else return (b2->score-b1->score);
845     }
846    
847    
848    
849 gpertea 93 GXAlnInfo* GreedyAlignRegion(const char* q_seq, int q_alnstart, int q_max,
850 gpertea 101 const char* s_seq, int s_alnstart, int s_max,
851     int reward, int penalty, int xdrop, CGreedyAlignData* gxmem,
852     CAlnTrim* trim, bool editscript) {
853 gpertea 93 GXEditScript* ed_script_fwd = NULL;
854     GXEditScript* ed_script_rev = NULL;
855     if ( q_alnstart>q_max || q_alnstart<1 || s_alnstart>s_max || s_alnstart<1 )
856     GError("GreedyAlign() Error: invalid anchor coordinate.\n");
857     q_alnstart--;
858     s_alnstart--;
859     if (q_seq==NULL || q_seq[0]==0 || s_seq==NULL || s_seq[0]==0)
860     GError("GreedyAlign() Error: attempt to use an empty sequence string!\n");
861     if (q_seq[q_alnstart]!=s_seq[s_alnstart])
862     GError("GreedyAlign() Error: improper anchor (mismatch)!\n");
863     int q_ext_l=0, q_ext_r=0, s_ext_l=0, s_ext_r=0;
864     const char* q=q_seq+q_alnstart;
865     int q_avail=q_max-q_alnstart;
866     const char* s=s_seq+s_alnstart;
867     int s_avail=s_max-s_alnstart;
868     if (penalty<0) penalty=-penalty;
869 gpertea 101 int MIN_GREEDY_SCORE=5*reward; //minimum score for an alignment to be reported for 0 diffs
870 gpertea 93 GXAlnInfo* alninfo=NULL;
871     bool freeAlnMem=(gxmem==NULL);
872     if (freeAlnMem) {
873 gpertea 101 gxmem=new CGreedyAlignData(reward, penalty, xdrop);
874 gpertea 93 }
875     else gxmem->reset();
876     int retscore = 0;
877     int numdiffs = 0;
878 gpertea 101 if (trim!=NULL && trim->type==galn_TrimLeft) {
879 gpertea 176 //intent: trimming the left side
880 gpertea 93 if (editscript)
881     ed_script_rev=new GXEditScript();
882    
883 gpertea 101 int numdiffs_l = GXGreedyExtend(s_seq, s_alnstart, q_seq, q_alnstart, true, xdrop,
884 gpertea 93 reward, penalty, s_ext_l, q_ext_l, *gxmem, ed_script_rev);
885     //check this extension here and bail out if it's not a good extension
886 gpertea 176 if (s_ext_l+(trim->seedlen>>1) < trim->safelen && s_alnstart+1-s_ext_l>trim->boundary) {
887 gpertea 93 delete ed_script_rev;
888     if (freeAlnMem) delete gxmem;
889     return NULL;
890     }
891    
892     if (editscript)
893     ed_script_fwd=new GXEditScript();
894     int numdiffs_r = GXGreedyExtend(s, s_avail, q, q_avail, false, xdrop,
895     reward, penalty, s_ext_r, q_ext_r, *gxmem, ed_script_fwd);
896     numdiffs=numdiffs_r+numdiffs_l;
897     //convert num diffs to actual score
898 gpertea 101 retscore = (q_ext_r + s_ext_r + q_ext_l + s_ext_l)*reward/2 - numdiffs*(reward+penalty);
899 gpertea 93 if (editscript)
900     ed_script_rev->Append(ed_script_fwd); //combine the two extensions
901     }
902     else {
903     if (editscript) {
904     ed_script_fwd=new GXEditScript();
905     }
906     int numdiffs_r = GXGreedyExtend(s, s_avail, q, q_avail, false, xdrop,
907     reward, penalty, s_ext_r, q_ext_r, *gxmem, ed_script_fwd);
908     //check extension here and bail out if not a good right extension
909     //assuming s_max is really at the right end of s_seq
910 gpertea 109 if (trim!=NULL && trim->type==galn_TrimRight &&
911 gpertea 176 s_ext_r+(trim->seedlen>>1) < trim->safelen &&
912     s_alnstart+s_ext_r<trim->boundary) {
913 gpertea 93 delete ed_script_fwd;
914     if (freeAlnMem) delete gxmem;
915     return NULL;
916     }
917     if (editscript)
918     ed_script_rev=new GXEditScript();
919     int numdiffs_l = GXGreedyExtend(s_seq, s_alnstart, q_seq, q_alnstart, true, xdrop,
920     reward, penalty, s_ext_l, q_ext_l, *gxmem, ed_script_rev);
921     //convert num diffs to actual score
922     numdiffs=numdiffs_r+numdiffs_l;
923 gpertea 101 retscore = (q_ext_r + s_ext_r + q_ext_l + s_ext_l)*reward/2 - numdiffs*(reward+penalty);
924 gpertea 93 if (editscript)
925     ed_script_rev->Append(ed_script_fwd); //combine the two extensions
926     }
927    
928     if (retscore>=MIN_GREEDY_SCORE) {
929     alninfo=new GXAlnInfo(q_seq, q_alnstart+1-q_ext_l,q_alnstart+q_ext_r, s_seq, s_alnstart+1-s_ext_l, s_alnstart+s_ext_r);
930     int hsp_length = GMIN(q_ext_l+q_ext_r, s_ext_l+s_ext_r);
931     alninfo->score=retscore;
932     alninfo->pid = 100 * (1 - ((double) numdiffs) / hsp_length);
933     #ifdef GDEBUG
934 gpertea 104 //if (ed_script_rev) {
935     // GMessage("Final Edit script ::: ");
936     // printEditScript(ed_script_rev);
937     // }
938 gpertea 93 #endif
939     alninfo->editscript=ed_script_rev;
940     alninfo->gapinfo = new CAlnGapInfo(ed_script_rev, alninfo->ql-1, alninfo->sl-1);
941     }
942     else {
943     if (freeAlnMem) delete gxmem;
944     delete ed_script_rev;
945     delete alninfo;
946     alninfo=NULL;
947     }
948     delete ed_script_fwd;
949     return alninfo;
950     }
951 gpertea 96
952 gpertea 101 GXAlnInfo* GreedyAlignRegion(const char* q_seq, int q_alnstart, int q_max,
953     const char* s_seq, int s_alnstart, int s_max, CGreedyAlignData* gxmem,
954     CAlnTrim* trim, bool editscript) {
955     int reward=2;
956     int penalty=3;
957     int xdrop=8;
958     if (gxmem) {
959     reward=gxmem->match_reward;
960     penalty=gxmem->mismatch_penalty;
961     xdrop=gxmem->x_drop;
962     }
963     return GreedyAlignRegion(q_seq, q_alnstart, q_max, s_seq, s_alnstart, s_max,
964     reward, penalty, xdrop, gxmem, trim, editscript);
965     }
966    
967 gpertea 176 GXAlnInfo* match_RightEnd(GXSeqData& sd, CGreedyAlignData* gxmem,
968     int min_pid) {
969 gpertea 96 bool editscript=false;
970     #ifdef GDEBUG
971     editscript=true;
972 gpertea 176 GMessage("==========> matching Right (3') end : %s\n", sd.aseq);
973 gpertea 96 #endif
974 gpertea 176 CAlnTrim trimInfo(galn_TrimRight, sd.bseq, sd.blen, sd.amlen);
975     GList<GXSeed> rseeds(true,true,false);
976 gpertea 101
977 gpertea 176 GXBandSet* alnbands=collectSeeds_R(rseeds, sd);
978 gpertea 101 GList<GXSeed> anchor_seeds(cmpSeedDiag, NULL, true); //stores unique seeds per diagonal
979 gpertea 99 //did we find a shortcut?
980     if (alnbands->qmatch) {
981 gpertea 174 #ifdef GDEBUG
982     GMessage("::: Found a quick long match at %d, len %d\n",
983     alnbands->qmatch->b_ofs, alnbands->qmatch->len);
984     #endif
985 gpertea 101 anchor_seeds.Add(alnbands->qmatch);
986     }
987     else {
988     int max_top_bands=5;
989     int top_band_count=0;
990     for (int b=0;b<alnbands->Count();b++) {
991 gpertea 174 if (alnbands->Get(b)->score<6) break;
992 gpertea 104 //#ifdef GDEBUG
993     //GMessage("\tBand %d score: %d\n", b, alnbands->Get(b)->score);
994     //#endif
995 gpertea 101 top_band_count++;
996     GXBand& band=*(alnbands->Get(b));
997     band.seeds.setSorted(cmpSeedScore);
998     anchor_seeds.Add(band.seeds.First());
999     band.tested=true;
1000     if (anchor_seeds.Count()>2 || top_band_count>max_top_bands) break;
1001     }
1002 gpertea 104 //#ifdef GDEBUG
1003     //GMessage("::: Collected %d anchor seeds.\n",anchor_seeds.Count());
1004     //#endif
1005 gpertea 101 }
1006 gpertea 96 GList<GXAlnInfo> galns(true,true,false);
1007     for (int i=0;i<anchor_seeds.Count();i++) {
1008     GXSeed& aseed=*anchor_seeds[i];
1009     int a1=aseed.a_ofs+(aseed.len>>1)+1;
1010     int a2=aseed.b_ofs+(aseed.len>>1)+1;
1011 gpertea 176 trimInfo.seedlen=aseed.len;
1012     GXAlnInfo* alninfo=GreedyAlignRegion(sd.aseq, a1, sd.alen,
1013     sd.bseq, a2, sd.blen, gxmem, &trimInfo, editscript);
1014 gpertea 174 if (alninfo && alninfo->pid>=min_pid &&
1015 gpertea 176 trimInfo.validate(alninfo->sl, alninfo->sr, alninfo->pid, alninfo->ql-1))
1016 gpertea 96 galns.AddIfNew(alninfo, true);
1017     else delete alninfo;
1018     }
1019 gpertea 174 if (galns.Count()==0 && alnbands->tmatch) {
1020     //last resort seed
1021     GXSeed& aseed=*alnbands->tmatch;
1022 gpertea 176 int halfseed=aseed.len>>1;
1023     int a1=aseed.a_ofs+halfseed+1;
1024     int a2=aseed.b_ofs+halfseed+1;
1025     trimInfo.seedlen=aseed.len;
1026     GXAlnInfo* alninfo=GreedyAlignRegion(sd.aseq, a1, sd.alen,
1027     sd.bseq, a2, sd.blen, gxmem, &trimInfo, editscript);
1028 gpertea 174 if (alninfo && alninfo->pid>=min_pid &&
1029 gpertea 176 trimInfo.validate(alninfo->sl, alninfo->sr, alninfo->pid, alninfo->ql-1))
1030 gpertea 174 galns.AddIfNew(alninfo, true);
1031     else delete alninfo;
1032     }
1033    
1034     /*
1035 gpertea 104 //special 3' end case: due to the seed scoring scheme being biased towards the 5' end of the read,
1036     //we should also try some seeds closer to the 3' end
1037 gpertea 101 if (galns.Count()==0) {
1038 gpertea 104 anchor_seeds.Clear();
1039 gpertea 101 alnbands->setSorted(cmpDiagBands_R);
1040     int max_top_bands=4;
1041     int top_band_count=0;
1042 gpertea 104 //#ifdef GDEBUG
1043     //GMessage(":::>> Retrying adjusting sort order.\n");
1044     //#endif
1045     if (alnbands->tmatch) {
1046     //anchor_seeds.setSorted(false);
1047     anchor_seeds.Add(alnbands->tmatch);
1048     }
1049 gpertea 101 for (int b=0;b<alnbands->Count();b++) {
1050     if (alnbands->Get(b)->score<4) break;
1051 gpertea 104 //#ifdef GDEBUG
1052     //GMessage("\tBand %d score: %d\n", b, alnbands->Get(b)->score);
1053     //#endif
1054 gpertea 101 if (alnbands->Get(b)->tested) continue;
1055     top_band_count++;
1056     GXBand& band=*(alnbands->Get(b));
1057     band.seeds.setSorted(cmpSeedScore);
1058     anchor_seeds.Add(band.seeds.First());
1059     if (anchor_seeds.Count()>2 || top_band_count>max_top_bands) break;
1060     }
1061 gpertea 104 //#ifdef GDEBUG
1062     //GMessage("::: Collected %d anchor seeds.\n",anchor_seeds.Count());
1063     //#endif
1064 gpertea 101 for (int i=0;i<anchor_seeds.Count();i++) {
1065     GXSeed& aseed=*anchor_seeds[i];
1066     int a1=aseed.a_ofs+(aseed.len>>1)+1;
1067     int a2=aseed.b_ofs+(aseed.len>>1)+1;
1068     GXAlnInfo* alninfo=GreedyAlignRegion(seqa, a1, seqa_len,
1069     seqb, a2, seqb_len, gxmem, &trimInfo, editscript);
1070     if (alninfo && alninfo->pid>=min_pid && trimInfo.validate(alninfo->sl, alninfo->sr, alninfo->pid, alninfo->ql-1))
1071     galns.AddIfNew(alninfo, true);
1072     else delete alninfo;
1073     }
1074 gpertea 174 } */
1075    
1076 gpertea 96 //---- done
1077     delete alnbands;
1078     if (galns.Count()) {
1079     GXAlnInfo* bestaln=galns.Shift();
1080 gpertea 104 #ifdef GDEBUG
1081     GMessage("Best alignment: a(%d..%d) align to b(%d..%d), score=%d, pid=%4.2f\n", bestaln->ql, bestaln->qr,
1082     bestaln->sl, bestaln->sr, bestaln->score, bestaln->pid);
1083     if (bestaln->gapinfo!=NULL) {
1084 gpertea 176 bestaln->gapinfo->printAlignment(stderr, sd.aseq, sd.alen, sd.bseq, sd.blen);
1085 gpertea 104 }
1086     #endif
1087    
1088 gpertea 96 return bestaln;
1089     }
1090     else return NULL;
1091     }
1092    
1093 gpertea 176 GXAlnInfo* match_LeftEnd(GXSeqData& sd, CGreedyAlignData* gxmem, int min_pid) {
1094 gpertea 96 bool editscript=false;
1095     #ifdef GDEBUG
1096     editscript=true;
1097 gpertea 176 GMessage("==========> matching Left (5') end : %s\n", sd.aseq);
1098 gpertea 96 #endif
1099 gpertea 176 CAlnTrim trimInfo(galn_TrimLeft, sd.bseq, sd.blen, sd.amlen);
1100 gpertea 96 GList<GXSeed> rseeds(true,true,false);
1101 gpertea 176 GXBandSet* alnbands=collectSeeds_L(rseeds, sd);
1102 gpertea 96 GList<GXSeed> anchor_seeds(cmpSeedDiag, NULL, true); //stores unique seeds per diagonal
1103 gpertea 101 if (alnbands->qmatch) {
1104 gpertea 174 #ifdef GDEBUG
1105     GMessage("::: Found a quick long match at %d, len %d\n",
1106     alnbands->qmatch->b_ofs, alnbands->qmatch->len);
1107     #endif
1108 gpertea 101 anchor_seeds.Add(alnbands->qmatch);
1109     }
1110     else {
1111     int max_top_bands=5;
1112     int top_band_count=0;
1113     for (int b=0;b<alnbands->Count();b++) {
1114 gpertea 174 if (alnbands->Get(b)->score<6) break;
1115 gpertea 104 //#ifdef GDEBUG
1116     //GMessage("\tBand %d score: %d\n", b, alnbands->Get(b)->score);
1117     //#endif
1118 gpertea 101 top_band_count++;
1119     GXBand& band=*(alnbands->Get(b));
1120     band.seeds.setSorted(cmpSeedScore);
1121     anchor_seeds.Add(band.seeds.First());
1122 gpertea 174 band.tested=true;
1123 gpertea 101 if (anchor_seeds.Count()>2 || top_band_count>max_top_bands) break;
1124     }
1125 gpertea 104 //#ifdef GDEBUG
1126     //GMessage("::: Collected %d anchor seeds.\n",anchor_seeds.Count());
1127     //#endif
1128 gpertea 101 }
1129 gpertea 174 GList<GXAlnInfo> galns(true,true,false);
1130     for (int i=0;i<anchor_seeds.Count();i++) {
1131 gpertea 96 GXSeed& aseed=*anchor_seeds[i];
1132     int a1=aseed.a_ofs+(aseed.len>>1)+1;
1133     int a2=aseed.b_ofs+(aseed.len>>1)+1;
1134 gpertea 176 trimInfo.seedlen=aseed.len;
1135     GXAlnInfo* alninfo=GreedyAlignRegion(sd.aseq, a1, sd.alen,
1136     sd.bseq, a2, sd.blen, gxmem, &trimInfo, editscript);
1137 gpertea 101 if (alninfo && alninfo->pid>=min_pid
1138 gpertea 176 && trimInfo.validate(alninfo->sl, alninfo->sr,
1139     alninfo->pid, sd.alen-alninfo->qr))
1140 gpertea 96 galns.AddIfNew(alninfo, true);
1141     else delete alninfo;
1142     }
1143 gpertea 104 if (galns.Count()==0 && alnbands->tmatch) {
1144 gpertea 174 //last resort seed
1145 gpertea 104 GXSeed& aseed=*alnbands->tmatch;
1146     int a1=aseed.a_ofs+(aseed.len>>1)+1;
1147     int a2=aseed.b_ofs+(aseed.len>>1)+1;
1148 gpertea 176 trimInfo.seedlen=aseed.len;
1149     GXAlnInfo* alninfo=GreedyAlignRegion(sd.aseq, a1, sd.alen,
1150     sd.bseq, a2, sd.blen, gxmem, &trimInfo, editscript);
1151 gpertea 174 if (alninfo && alninfo->pid>=min_pid &&
1152 gpertea 176 trimInfo.validate(alninfo->sl, alninfo->sr, alninfo->pid, sd.alen-alninfo->qr))
1153 gpertea 174 galns.Add(alninfo);
1154 gpertea 104 }
1155 gpertea 174 /*
1156 gpertea 96 #ifdef GDEBUG
1157 gpertea 174 //print valid alignments found
1158 gpertea 96 for (int i=0;i<galns.Count();i++) {
1159     GXAlnInfo* alninfo=galns[i];
1160     GMessage("a(%d..%d) align to b(%d..%d), score=%d, pid=%4.2f\n", alninfo->ql, alninfo->qr,
1161     alninfo->sl, alninfo->sr, alninfo->score, alninfo->pid);
1162     if (alninfo->gapinfo!=NULL) {
1163     GMessage("Alignment:\n");
1164     alninfo->gapinfo->printAlignment(stderr, seqa, seqa_len, seqb, seqb_len);
1165     }
1166     }
1167 gpertea 174 #endif
1168 gpertea 104 */
1169 gpertea 96 //---- done
1170     delete alnbands;
1171     if (galns.Count()) {
1172     GXAlnInfo* bestaln=galns.Shift();
1173 gpertea 104 #ifdef GDEBUG
1174     GMessage("Best alignment: a(%d..%d) align to b(%d..%d), score=%d, pid=%4.2f\n", bestaln->ql, bestaln->qr,
1175     bestaln->sl, bestaln->sr, bestaln->score, bestaln->pid);
1176     if (bestaln->gapinfo!=NULL) {
1177 gpertea 176 bestaln->gapinfo->printAlignment(stderr, sd.aseq, sd.alen,
1178     sd.bseq, sd.blen);
1179 gpertea 104 }
1180     #endif
1181 gpertea 96 return bestaln;
1182     }
1183     else return NULL;
1184     }