#include #include #include #include #include #include #include #ifdef _WIN32 #include #endif #include "kstring.h" #include "bam.h" #include "sam_header.h" #include "kseq.h" #include "khash.h" KSTREAM_INIT(gzFile, gzread, 16384) KHASH_MAP_INIT_STR(ref, uint64_t) void bam_init_header_hash(bam_header_t *header); void bam_destroy_header_hash(bam_header_t *header); int32_t bam_get_tid(const bam_header_t *header, const char *seq_name); unsigned char bam_nt16_table[256] = { 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 1, 2, 4, 8, 15,15,15,15, 15,15,15,15, 15, 0 /*=*/,15,15, 15, 1,14, 2, 13,15,15, 4, 11,15,15,12, 15, 3,15,15, 15,15, 5, 6, 8,15, 7, 9, 15,10,15,15, 15,15,15,15, 15, 1,14, 2, 13,15,15, 4, 11,15,15,12, 15, 3,15,15, 15,15, 5, 6, 8,15, 7, 9, 15,10,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15, 15,15,15,15 }; unsigned short bam_char2flag_table[256] = { 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,BAM_FREAD1,BAM_FREAD2,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, BAM_FPROPER_PAIR,0,BAM_FMREVERSE,0, 0,BAM_FMUNMAP,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, BAM_FDUP,0,BAM_FQCFAIL,0, 0,0,0,0, 0,0,0,0, BAM_FPAIRED,0,BAM_FREVERSE,BAM_FSECONDARY, 0,BAM_FUNMAP,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0, 0,0,0,0 }; char *bam_nt16_rev_table = "=ACMGRSVTWYHKDBN"; struct __tamFile_t { gzFile fp; kstream_t *ks; kstring_t *str; uint64_t n_lines; int is_first; }; char **__bam_get_lines(const char *fn, int *_n) // for bam_plcmd.c only { char **list = 0, *s; int n = 0, dret, m = 0; gzFile fp = (strcmp(fn, "-") == 0)? gzdopen(fileno(stdin), "r") : gzopen(fn, "r"); kstream_t *ks; kstring_t *str; str = (kstring_t*)calloc(1, sizeof(kstring_t)); ks = ks_init(fp); while (ks_getuntil(ks, '\n', str, &dret) > 0) { if (n == m) { m = m? m << 1 : 16; list = (char**)realloc(list, m * sizeof(char*)); } if (str->s[str->l-1] == '\r') str->s[--str->l] = '\0'; s = list[n++] = (char*)calloc(str->l + 1, 1); strcpy(s, str->s); } ks_destroy(ks); gzclose(fp); free(str->s); free(str); *_n = n; return list; } static bam_header_t *hash2header(const kh_ref_t *hash) { bam_header_t *header; khiter_t k; header = bam_header_init(); header->n_targets = kh_size(hash); header->target_name = (char**)calloc(kh_size(hash), sizeof(char*)); header->target_len = (uint32_t*)calloc(kh_size(hash), 4); for (k = kh_begin(hash); k != kh_end(hash); ++k) { if (kh_exist(hash, k)) { int i = (int)kh_value(hash, k); header->target_name[i] = (char*)kh_key(hash, k); header->target_len[i] = kh_value(hash, k)>>32; } } bam_init_header_hash(header); return header; } bam_header_t *sam_header_read2(const char *fn) { bam_header_t *header; int c, dret, ret, error = 0; gzFile fp; kstream_t *ks; kstring_t *str; kh_ref_t *hash; khiter_t k; if (fn == 0) return 0; fp = (strcmp(fn, "-") == 0)? gzdopen(fileno(stdin), "r") : gzopen(fn, "r"); if (fp == 0) return 0; hash = kh_init(ref); ks = ks_init(fp); str = (kstring_t*)calloc(1, sizeof(kstring_t)); while (ks_getuntil(ks, 0, str, &dret) > 0) { char *s = strdup(str->s); int len, i; i = kh_size(hash); ks_getuntil(ks, 0, str, &dret); len = atoi(str->s); k = kh_put(ref, hash, s, &ret); if (ret == 0) { fprintf(stderr, "[sam_header_read2] duplicated sequence name: %s\n", s); error = 1; } kh_value(hash, k) = (uint64_t)len<<32 | i; if (dret != '\n') while ((c = ks_getc(ks)) != '\n' && c != -1); } ks_destroy(ks); gzclose(fp); free(str->s); free(str); fprintf(stderr, "[sam_header_read2] %d sequences loaded.\n", kh_size(hash)); if (error) return 0; header = hash2header(hash); kh_destroy(ref, hash); return header; } static inline uint8_t *alloc_data(bam1_t *b, int size) { if (b->m_data < size) { b->m_data = size; kroundup32(b->m_data); b->data = (uint8_t*)realloc(b->data, b->m_data); } return b->data; } static inline void parse_error(int64_t n_lines, const char * __restrict msg) { fprintf(stderr, "Parse error at line %lld: %s\n", (long long)n_lines, msg); abort(); } static inline void append_text(bam_header_t *header, kstring_t *str) { size_t x = header->l_text, y = header->l_text + str->l + 2; // 2 = 1 byte dret + 1 byte null kroundup32(x); kroundup32(y); if (x < y) { header->n_text = y; header->text = (char*)realloc(header->text, y); if ( !header->text ) { fprintf(stderr,"realloc failed to alloc %ld bytes\n", y); abort(); } } // Sanity check if ( header->l_text+str->l+1 >= header->n_text ) { fprintf(stderr,"append_text FIXME: %ld>=%ld, x=%ld,y=%ld\n", header->l_text+str->l+1,header->n_text,x,y); abort(); } strncpy(header->text + header->l_text, str->s, str->l+1); // we cannot use strcpy() here. header->l_text += str->l + 1; header->text[header->l_text] = 0; } int sam_header_parse(bam_header_t *h) { char **tmp; int i; free(h->target_len); free(h->target_name); h->n_targets = 0; h->target_len = 0; h->target_name = 0; if (h->l_text < 3) return 0; if (h->dict == 0) h->dict = sam_header_parse2(h->text); tmp = sam_header2list(h->dict, "SQ", "SN", &h->n_targets); if (h->n_targets == 0) return 0; h->target_name = calloc(h->n_targets, sizeof(void*)); for (i = 0; i < h->n_targets; ++i) h->target_name[i] = strdup(tmp[i]); free(tmp); tmp = sam_header2list(h->dict, "SQ", "LN", &h->n_targets); h->target_len = calloc(h->n_targets, 4); for (i = 0; i < h->n_targets; ++i) h->target_len[i] = atoi(tmp[i]); free(tmp); return h->n_targets; } bam_header_t *sam_header_read(tamFile fp) { int ret, dret; bam_header_t *header = bam_header_init(); kstring_t *str = fp->str; while ((ret = ks_getuntil(fp->ks, KS_SEP_TAB, str, &dret)) >= 0 && str->s[0] == '@') { // skip header str->s[str->l] = dret; // note that str->s is NOT null terminated!! append_text(header, str); if (dret != '\n') { ret = ks_getuntil(fp->ks, '\n', str, &dret); str->s[str->l] = '\n'; // NOT null terminated!! append_text(header, str); } ++fp->n_lines; } sam_header_parse(header); bam_init_header_hash(header); fp->is_first = 1; return header; } int sam_read1(tamFile fp, bam_header_t *header, bam1_t *b) { int ret, doff, doff0, dret, z = 0; bam1_core_t *c = &b->core; kstring_t *str = fp->str; kstream_t *ks = fp->ks; if (fp->is_first) { fp->is_first = 0; ret = str->l; } else { do { // special consideration for empty lines ret = ks_getuntil(fp->ks, KS_SEP_TAB, str, &dret); if (ret >= 0) z += str->l + 1; } while (ret == 0); } if (ret < 0) return -1; ++fp->n_lines; doff = 0; { // name c->l_qname = strlen(str->s) + 1; memcpy(alloc_data(b, doff + c->l_qname) + doff, str->s, c->l_qname); doff += c->l_qname; } { // flag long flag; char *s; ret = ks_getuntil(ks, KS_SEP_TAB, str, &dret); z += str->l + 1; flag = strtol((char*)str->s, &s, 0); if (*s) { // not the end of the string flag = 0; for (s = str->s; *s; ++s) flag |= bam_char2flag_table[(int)*s]; } c->flag = flag; } { // tid, pos, qual ret = ks_getuntil(ks, KS_SEP_TAB, str, &dret); z += str->l + 1; c->tid = bam_get_tid(header, str->s); if (c->tid < 0 && strcmp(str->s, "*")) { if (header->n_targets == 0) { fprintf(stderr, "[sam_read1] missing header? Abort!\n"); exit(1); } else fprintf(stderr, "[sam_read1] reference '%s' is recognized as '*'.\n", str->s); } ret = ks_getuntil(ks, KS_SEP_TAB, str, &dret); z += str->l + 1; c->pos = isdigit(str->s[0])? atoi(str->s) - 1 : -1; ret = ks_getuntil(ks, KS_SEP_TAB, str, &dret); z += str->l + 1; c->qual = isdigit(str->s[0])? atoi(str->s) : 0; if (ret < 0) return -2; } { // cigar char *s, *t; int i, op; long x; c->n_cigar = 0; if (ks_getuntil(ks, KS_SEP_TAB, str, &dret) < 0) return -3; z += str->l + 1; if (str->s[0] != '*') { for (s = str->s; *s; ++s) { if ((isalpha(*s)) || (*s=='=')) ++c->n_cigar; else if (!isdigit(*s)) parse_error(fp->n_lines, "invalid CIGAR character"); } b->data = alloc_data(b, doff + c->n_cigar * 4); for (i = 0, s = str->s; i != c->n_cigar; ++i) { x = strtol(s, &t, 10); op = toupper(*t); if (op == 'M') op = BAM_CMATCH; else if (op == 'I') op = BAM_CINS; else if (op == 'D') op = BAM_CDEL; else if (op == 'N') op = BAM_CREF_SKIP; else if (op == 'S') op = BAM_CSOFT_CLIP; else if (op == 'H') op = BAM_CHARD_CLIP; else if (op == 'P') op = BAM_CPAD; else if (op == '=') op = BAM_CEQUAL; else if (op == 'X') op = BAM_CDIFF; else parse_error(fp->n_lines, "invalid CIGAR operation"); s = t + 1; bam1_cigar(b)[i] = x << BAM_CIGAR_SHIFT | op; } if (*s) parse_error(fp->n_lines, "unmatched CIGAR operation"); c->bin = bam_reg2bin(c->pos, bam_calend(c, bam1_cigar(b))); doff += c->n_cigar * 4; } else { if (!(c->flag&BAM_FUNMAP)) { fprintf(stderr, "Parse warning at line %lld: mapped sequence without CIGAR\n", (long long)fp->n_lines); c->flag |= BAM_FUNMAP; } c->bin = bam_reg2bin(c->pos, c->pos + 1); } } { // mtid, mpos, isize ret = ks_getuntil(ks, KS_SEP_TAB, str, &dret); z += str->l + 1; c->mtid = strcmp(str->s, "=")? bam_get_tid(header, str->s) : c->tid; ret = ks_getuntil(ks, KS_SEP_TAB, str, &dret); z += str->l + 1; c->mpos = isdigit(str->s[0])? atoi(str->s) - 1 : -1; ret = ks_getuntil(ks, KS_SEP_TAB, str, &dret); z += str->l + 1; c->isize = (str->s[0] == '-' || isdigit(str->s[0]))? atoi(str->s) : 0; if (ret < 0) return -4; } { // seq and qual int i; uint8_t *p = 0; if (ks_getuntil(ks, KS_SEP_TAB, str, &dret) < 0) return -5; // seq z += str->l + 1; if (strcmp(str->s, "*")) { c->l_qseq = strlen(str->s); if (c->n_cigar && c->l_qseq != (int32_t)bam_cigar2qlen(c, bam1_cigar(b))) { fprintf(stderr, "Line %ld, sequence length %i vs %i from CIGAR\n", (long)fp->n_lines, c->l_qseq, (int32_t)bam_cigar2qlen(c, bam1_cigar(b))); parse_error(fp->n_lines, "CIGAR and sequence length are inconsistent"); } p = (uint8_t*)alloc_data(b, doff + c->l_qseq + (c->l_qseq+1)/2) + doff; memset(p, 0, (c->l_qseq+1)/2); for (i = 0; i < c->l_qseq; ++i) p[i/2] |= bam_nt16_table[(int)str->s[i]] << 4*(1-i%2); } else c->l_qseq = 0; if (ks_getuntil(ks, KS_SEP_TAB, str, &dret) < 0) return -6; // qual z += str->l + 1; if (strcmp(str->s, "*") && c->l_qseq != strlen(str->s)) parse_error(fp->n_lines, "sequence and quality are inconsistent"); p += (c->l_qseq+1)/2; if (strcmp(str->s, "*") == 0) for (i = 0; i < c->l_qseq; ++i) p[i] = 0xff; else for (i = 0; i < c->l_qseq; ++i) p[i] = str->s[i] - 33; doff += c->l_qseq + (c->l_qseq+1)/2; } doff0 = doff; if (dret != '\n' && dret != '\r') { // aux while (ks_getuntil(ks, KS_SEP_TAB, str, &dret) >= 0) { uint8_t *s, type, key[2]; z += str->l + 1; if (str->l < 6 || str->s[2] != ':' || str->s[4] != ':') parse_error(fp->n_lines, "missing colon in auxiliary data"); key[0] = str->s[0]; key[1] = str->s[1]; type = str->s[3]; s = alloc_data(b, doff + 3) + doff; s[0] = key[0]; s[1] = key[1]; s += 2; doff += 2; if (type == 'A' || type == 'a' || type == 'c' || type == 'C') { // c and C for backward compatibility s = alloc_data(b, doff + 2) + doff; *s++ = 'A'; *s = str->s[5]; doff += 2; } else if (type == 'I' || type == 'i') { long long x; s = alloc_data(b, doff + 5) + doff; x = (long long)atoll(str->s + 5); if (x < 0) { if (x >= -127) { *s++ = 'c'; *(int8_t*)s = (int8_t)x; s += 1; doff += 2; } else if (x >= -32767) { *s++ = 's'; *(int16_t*)s = (int16_t)x; s += 2; doff += 3; } else { *s++ = 'i'; *(int32_t*)s = (int32_t)x; s += 4; doff += 5; if (x < -2147483648ll) fprintf(stderr, "Parse warning at line %lld: integer %lld is out of range.", (long long)fp->n_lines, x); } } else { if (x <= 255) { *s++ = 'C'; *s++ = (uint8_t)x; doff += 2; } else if (x <= 65535) { *s++ = 'S'; *(uint16_t*)s = (uint16_t)x; s += 2; doff += 3; } else { *s++ = 'I'; *(uint32_t*)s = (uint32_t)x; s += 4; doff += 5; if (x > 4294967295ll) fprintf(stderr, "Parse warning at line %lld: integer %lld is out of range.", (long long)fp->n_lines, x); } } } else if (type == 'f') { s = alloc_data(b, doff + 5) + doff; *s++ = 'f'; *(float*)s = (float)atof(str->s + 5); s += 4; doff += 5; } else if (type == 'd') { s = alloc_data(b, doff + 9) + doff; *s++ = 'd'; *(float*)s = (float)atof(str->s + 9); s += 8; doff += 9; } else if (type == 'Z' || type == 'H') { int size = 1 + (str->l - 5) + 1; if (type == 'H') { // check whether the hex string is valid int i; if ((str->l - 5) % 2 == 1) parse_error(fp->n_lines, "length of the hex string not even"); for (i = 0; i < str->l - 5; ++i) { int c = toupper(str->s[5 + i]); if (!((c >= '0' && c <= '9') || (c >= 'A' && c <= 'F'))) parse_error(fp->n_lines, "invalid hex character"); } } s = alloc_data(b, doff + size) + doff; *s++ = type; memcpy(s, str->s + 5, str->l - 5); s[str->l - 5] = 0; doff += size; } else if (type == 'B') { int32_t n = 0, Bsize, k = 0, size; char *p; if (str->l < 8) parse_error(fp->n_lines, "too few values in aux type B"); Bsize = bam_aux_type2size(str->s[5]); // the size of each element for (p = (char*)str->s + 6; *p; ++p) // count the number of elements in the array if (*p == ',') ++n; p = str->s + 7; // now p points to the first number in the array size = 6 + Bsize * n; // total number of bytes allocated to this tag s = alloc_data(b, doff + 6 * Bsize * n) + doff; // allocate memory *s++ = 'B'; *s++ = str->s[5]; memcpy(s, &n, 4); s += 4; // write the number of elements if (str->s[5] == 'c') while (p < str->s + str->l) ((int8_t*)s)[k++] = (int8_t)strtol(p, &p, 0), ++p; else if (str->s[5] == 'C') while (p < str->s + str->l) ((uint8_t*)s)[k++] = (uint8_t)strtol(p, &p, 0), ++p; else if (str->s[5] == 's') while (p < str->s + str->l) ((int16_t*)s)[k++] = (int16_t)strtol(p, &p, 0), ++p; // FIXME: avoid unaligned memory else if (str->s[5] == 'S') while (p < str->s + str->l) ((uint16_t*)s)[k++] = (uint16_t)strtol(p, &p, 0), ++p; else if (str->s[5] == 'i') while (p < str->s + str->l) ((int32_t*)s)[k++] = (int32_t)strtol(p, &p, 0), ++p; else if (str->s[5] == 'I') while (p < str->s + str->l) ((uint32_t*)s)[k++] = (uint32_t)strtol(p, &p, 0), ++p; else if (str->s[5] == 'f') while (p < str->s + str->l) ((float*)s)[k++] = (float)strtod(p, &p), ++p; else parse_error(fp->n_lines, "unrecognized array type"); s += Bsize * n; doff += size; } else parse_error(fp->n_lines, "unrecognized type"); if (dret == '\n' || dret == '\r') break; } } b->l_aux = doff - doff0; b->data_len = doff; return z; } tamFile sam_open(const char *fn) { tamFile fp; gzFile gzfp = (strcmp(fn, "-") == 0)? gzdopen(fileno(stdin), "rb") : gzopen(fn, "rb"); if (gzfp == 0) return 0; fp = (tamFile)calloc(1, sizeof(struct __tamFile_t)); fp->str = (kstring_t*)calloc(1, sizeof(kstring_t)); fp->fp = gzfp; fp->ks = ks_init(fp->fp); return fp; } void sam_close(tamFile fp) { if (fp) { ks_destroy(fp->ks); gzclose(fp->fp); free(fp->str->s); free(fp->str); free(fp); } }