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#include "GBam.h"
#include <ctype.h>
#include "kstring.h"
#define _cigOp(c) ((c)&BAM_CIGAR_MASK)
#define _cigLen(c) ((c)>>BAM_CIGAR_SHIFT)
//auxiliary functions for low level BAM record creation
uint8_t* realloc_bdata(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);
}
if (b->data_len<size) b->data_len=size;
return b->data;
}
uint8_t* dupalloc_bdata(bam1_t *b, int size) {
//same as realloc_bdata, but does not free previous data
//but returns it instead
//it ALWAYS duplicates data
b->m_data = size;
kroundup32(b->m_data);
uint8_t* odata=b->data;
b->data = (uint8_t*)malloc(b->m_data);
memcpy((void*)b->data, (void*)odata, b->data_len);
b->data_len=size;
return odata; //user must FREE this after
}
GBamRecord::GBamRecord(const char* qname, int32_t gseq_tid,
int pos, bool reverse, const char* qseq,
const char* cigar, const char* quals):iflags(0), exons(1),
clipL(0), clipR(0), mapped_len(0), uval(0) {
novel=true;
bam_header=NULL;
b=bam_init1();
if (pos<=0 || gseq_tid<0) {
b->core.pos=-1; //unmapped
b->core.flag |= BAM_FUNMAP;
gseq_tid=-1;
}
else b->core.pos=pos-1; //BAM is 0-based
b->core.tid=gseq_tid;
b->core.qual=255;
b->core.mtid=-1;
b->core.mpos=-1;
int l_qseq=strlen(qseq);
//this may not be accurate, setting CIGAR is the correct way
//b->core.bin = bam_reg2bin(b->core.pos, b->core.pos+l_qseq-1);
b->core.l_qname=strlen(qname)+1; //includes the \0 at the end
memcpy(realloc_bdata(b, b->core.l_qname), qname, b->core.l_qname);
set_cigar(cigar); //this will also set core.bin
add_sequence(qseq, l_qseq);
add_quals(quals); //quals must be given as Phred33
if (reverse) { b->core.flag |= BAM_FREVERSE ; }
}
GBamRecord::GBamRecord(const char* qname, int32_t samflags, int32_t g_tid,
int pos, int map_qual, const char* cigar, int32_t mg_tid, int mate_pos,
int insert_size, const char* qseq, const char* quals,
GVec<char*>* aux_strings):iflags(0), exons(1), uval(0) {
novel=true;
bam_header=NULL;
b=bam_init1();
b->core.tid=g_tid;
b->core.pos = (pos<=0) ? -1 : pos-1; //BAM is 0-based
b->core.qual=map_qual;
int l_qseq=strlen(qseq);
b->core.l_qname=strlen(qname)+1; //includes the \0 at the end
memcpy(realloc_bdata(b, b->core.l_qname), qname, b->core.l_qname);
set_cigar(cigar); //this will also set core.bin
add_sequence(qseq, l_qseq);
add_quals(quals); //quals must be given as Phred33
set_flags(samflags);
set_mdata(mg_tid, (int32_t)(mate_pos-1), (int32_t)insert_size);
if (aux_strings!=NULL) {
for (int i=0;i<aux_strings->Count();i++) {
add_aux(aux_strings->Get(i));
}
}
}
void GBamRecord::set_cigar(const char* cigar) {
//requires b->core.pos and b->core.flag to have been set properly PRIOR to this call
int doff=b->core.l_qname;
uint8_t* after_cigar=NULL;
int after_cigar_len=0;
uint8_t* prev_bdata=NULL;
if (b->data_len>doff) {
//cigar string already allocated, replace it
int d=b->core.l_qname + b->core.n_cigar * 4;//offset of after-cigar data
after_cigar=b->data+d;
after_cigar_len=b->data_len-d;
}
const char *s;
char *t;
int i, op;
long x;
b->core.n_cigar = 0;
if (cigar != NULL && strcmp(cigar, "*") != 0) {
for (s = cigar; *s; ++s) {
if (isalpha(*s)) b->core.n_cigar++;
else if (!isdigit(*s)) {
GError("Error: invalid CIGAR character (%s)\n",cigar);
}
}
if (after_cigar_len>0) { //replace/insert into existing full data
prev_bdata=dupalloc_bdata(b, doff + b->core.n_cigar * 4 + after_cigar_len);
memcpy((void*)(b->data+doff+b->core.n_cigar*4),(void*)after_cigar, after_cigar_len);
free(prev_bdata);
}
else {
realloc_bdata(b, doff + b->core.n_cigar * 4);
}
for (i = 0, s = cigar; i != b->core.n_cigar; ++i) {
x = strtol(s, &t, 10);
op = toupper(*t);
if (op == 'M' || op == '=' || op == 'X') 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; //has_Introns=true;
else if (op == 'S') op = BAM_CSOFT_CLIP; //soft_Clipped=true;
else if (op == 'H') op = BAM_CHARD_CLIP; //hard_Clipped=true;
else if (op == 'P') op = BAM_CPAD;
else GError("Error: invalid CIGAR operation (%s)\n",cigar);
s = t + 1;
bam1_cigar(b)[i] = x << BAM_CIGAR_SHIFT | op;
}
if (*s) GError("Error: unmatched CIGAR operation (%s)\n",cigar);
b->core.bin = bam_reg2bin(b->core.pos, bam_calend(&b->core, bam1_cigar(b)));
} else {//no CIGAR string given
if (!(b->core.flag&BAM_FUNMAP)) {
GMessage("Warning: mapped sequence without CIGAR (%s)\n", (char*)b->data);
b->core.flag |= BAM_FUNMAP;
}
b->core.bin = bam_reg2bin(b->core.pos, b->core.pos + 1);
}
setupCoordinates();
} //set_cigar()
void GBamRecord::add_sequence(const char* qseq, int slen) {
//must be called AFTER set_cigar (cannot replace existing sequence for now)
if (qseq==NULL) return; //should we ever care about this?
if (slen<0) slen=strlen(qseq);
int doff = b->core.l_qname + b->core.n_cigar * 4;
if (strcmp(qseq, "*")!=0) {
b->core.l_qseq=slen;
if (b->core.n_cigar && b->core.l_qseq != (int32_t)bam_cigar2qlen(&b->core, bam1_cigar(b)))
GError("Error: CIGAR and sequence length are inconsistent!(%s)\n",
qseq);
uint8_t* p = (uint8_t*)realloc_bdata(b, doff + (b->core.l_qseq+1)/2 + b->core.l_qseq) + doff;
//also allocated quals memory
memset(p, 0, (b->core.l_qseq+1)/2);
for (int i = 0; i < b->core.l_qseq; ++i)
p[i/2] |= bam_nt16_table[(int)qseq[i]] << 4*(1-i%2);
} else b->core.l_qseq = 0;
}
void GBamRecord::add_quals(const char* quals) {
//requires core.l_qseq already set
//and must be called AFTER add_sequence(), which also allocates the memory for quals
uint8_t* p = b->data+(b->core.l_qname + b->core.n_cigar * 4 + (b->core.l_qseq+1)/2);
if (quals==NULL || strcmp(quals, "*") == 0) {
for (int i=0;i < b->core.l_qseq; i++) p[i] = 0xff;
return;
}
for (int i=0;i < b->core.l_qseq; i++) p[i] = quals[i]-33;
}
void GBamRecord::add_aux(const char* str) {
//requires: being called AFTER add_quals()
int strl=strlen(str);
//int doff = b->core.l_qname + b->core.n_cigar*4 + (b->core.l_qseq+1)/2 + b->core.l_qseq + b->l_aux;
//int doff0=doff;
if (strl < 6 || str[2] != ':' || str[4] != ':')
parse_error("missing colon in auxiliary data");
tag[0] = str[0]; tag[1] = str[1];
uint8_t atype = str[3];
uint8_t* adata=abuf;
int alen=0;
if (atype == 'A' || atype == 'a' || atype == 'c' || atype == 'C') { // c and C for backward compatibility
atype='A';
alen=1;
adata=(uint8_t*)&str[5];
}
else if (atype == 'I' || atype == 'i') {
long long x=strtoll(str+5, NULL, 10); //(long long)atoll(str + 5);
//long x=(long)atol(str + 5);
if (x < 0) {
if (x >= -127) {
atype='c';
abuf[0] = (int8_t)x;
alen=1;
}
else if (x >= -32767) {
atype = 's';
*(int16_t*)abuf = (int16_t)x;
alen=2;
}
else {
atype='i';
*(int32_t*)abuf = (int32_t)x;
alen=4;
if (x < -2147483648ll)
GMessage("Parse warning: integer %lld is out of range.", x);
}
} else { //x >=0
if (x <= 255) {
atype = 'C';
abuf[0] = (uint8_t)x;
alen=1;
}
else if (x <= 65535) {
atype='S';
*(uint16_t*)abuf = (uint16_t)x;
alen=2;
}
else {
atype='I';
*(uint32_t*)abuf = (uint32_t)x;
alen=4;
if (x > 4294967295ll)
GMessage("Parse warning: integer %lld is out of range.", x);
}
}
} //integer type
else if (atype == 'f') {
*(float*)abuf = (float)atof(str + 5);
alen = sizeof(float);
}
else if (atype == 'd') { //?
*(float*)abuf = (float)atof(str + 9);
alen=8;
}
else if (atype == 'Z' || atype == 'H') {
if (atype == 'H') { // check whether the hex string is valid
if ((strl - 5) % 2 == 1) parse_error("length of the hex string not even");
for (int i = 0; i < strl - 5; ++i) {
int c = toupper(str[5 + i]);
if (!((c >= '0' && c <= '9') || (c >= 'A' && c <= 'F')))
parse_error("invalid hex character");
}
}
memcpy(abuf, str + 5, strl - 5);
abuf[strl-5] = 0;
alen=strl-4;
} else parse_error("unrecognized aux type");
this->add_aux(tag, atype, alen, adata);
}//add_aux()
int interpret_CIGAR(char cop, int cl, int aln_start) {
// returns the number of bases "aligned" (matches or mismatches) from the read
// gpos = current genomic position (will end up as right coordinate on the genome)
// rpos = read position (will end up as the length of the read)
// cop = CIGAR operation, cl = operation length
int mbases = 0; //count "aligned" bases (includes mismatches)
int rpos = 0;
int gpos = aln_start;
int num_mismatches=0; //NM tag value = edit distance
switch (cop) {
case BAM_CDIFF: // X
num_mismatches+=cl;
case BAM_CMATCH: // M
//have to actually check for mismatches: num_mismatches+=count_mismatches;
case BAM_CEQUAL: // =
//printf("[%d-%d]", gpos, gpos + cl - 1);
gpos+=cl;
rpos+=cl;
++mbases;
break;
case BAM_CPAD:
// printf("[%d-%d]", pos, pos + cl - 1); // Spans positions, No Coverage
gpos+=cl;
break;
case BAM_CHARD_CLIP:
// printf("[%d]", pos); // No coverage
// gpos is not advanced by this operation
break;
case BAM_CSOFT_CLIP: // S
//soft clipped bases, present in SEQ
rpos+=cl;
break;
case BAM_CINS: // I
// No Coverage
// adds cl bases "throughput" but not genomic position "coverage" (gap in genomic seq)
// should also add cl to the number of "mismatches" (unaligned read bases)
num_mismatches+=cl;
// How you handle this is application dependent
// gpos is not advanced by this operation
rpos+=cl;
break;
case BAM_CDEL: // D
//deletion in reference sequence relative to the read (gap in read sequence)
// printf("[%d-%d]", pos, pos + cl - 1);
// Spans positions
num_mismatches+=cl;
gpos += cl;
break;
case BAM_CREF_SKIP: // N
// intron
//special skip operation, not contributing to "edit distance",
// printf("[%d-%d]", pos, pos + cl - 1); // Spans positions, No Coverage
// so num_mismatches is not updated
gpos+=cl;
break;
default:
fprintf(stderr, "Unhandled cigar_op %d:%d\n", cop, cl);
//printf("?");
}
return mbases;
} // interpret_CIGAR(), just a reference of CIGAR operations interpretation
void GBamRecord::setupCoordinates() {
const bam1_core_t *c = &b->core;
if (c->flag & BAM_FUNMAP) return; /* skip unmapped reads */
uint32_t *cigar = bam1_cigar(b);
//uint32_t *p = bam1_cigar(b);
//--- prevent alignment error here (reported by UB-sanitazer):
//uint32_t *cigar= new uint32_t[c->n_cigar];
//memcpy(cigar, p, c->n_cigar * sizeof(uint32_t));
//--- UBsan protection end
int l=0;
mapped_len=0;
clipL=0;
clipR=0;
start=c->pos+1; //genomic start coordinate, 1-based (BAM core.pos is 0-based)
GSeg exon;
int exstart=c->pos;
bool intron=false;
bool ins=false;
for (int i = 0; i < c->n_cigar; ++i) {
unsigned char op = _cigOp(cigar[i]);
switch(op) {
case BAM_CEQUAL: // =
case BAM_CDIFF: // X
case BAM_CMATCH: // M
case BAM_CDEL: // D
l+=_cigLen(cigar[i]);
intron=false;
ins=false;
break;
case BAM_CINS: // I
//rpos+=cl; //gpos not advanced
//take care of cases where there is an ins within an intron
ins=true;
break;
case BAM_CREF_SKIP: // N
//intron starts
//exon ends here
if(!ins || !intron) { // insertion in the middle of an intron --> adjust last exon
exon.start=exstart+1;
exon.end=c->pos+l;
exons.Add(exon);
mapped_len+=exon.len();
}
has_Introns=true;
l += _cigLen(cigar[i]);
exstart=c->pos+l;
intron=true;
break;
case BAM_CSOFT_CLIP: // S
soft_Clipped=true;
if (l) clipR=_cigLen(cigar[i]);
else clipL=_cigLen(cigar[i]);
intron=false; ins=false;
break;
case BAM_CHARD_CLIP:
hard_Clipped=true;
intron=false; ins=false;
break;
case BAM_CPAD:
//gpos+=cl;
intron=false; ins=false; //?
break;
default:
int cl=_cigLen(cigar[i]);
fprintf(stderr, "Unhandled CIGAR operation %d:%d\n", op, cl);
}
}
exon.start=exstart+1;
exon.end=c->pos+l;
exons.Add(exon);
mapped_len+=exon.len();
end=exon.end; //genomic end coordinate
}
uint8_t* GBamRecord::find_tag(const char tag[2]) {
return bam_aux_get(this->b, tag);
}
char GBamRecord::tag_char(const char tag[2]) { //retrieve tag data as single char
uint8_t* s=find_tag(tag);
if (s) return ( bam_aux2A(s) );
return 0;
}
char GBamRecord::tag_char1(const char tag[2]) { //just the first char from Z type tags
uint8_t* s=bam_aux_get(this->b, tag);
if (s==NULL) return 0;
int type;
type = *s++;
if (s == 0) return 0;
if (type == 'A' || type == 'Z') return *(char*)s;
else return 0;
}
int GBamRecord::tag_int(const char tag[2]) { //get the numeric value of tag
uint8_t *s=find_tag(tag);
if (s) return ( bam_aux2i(s) );
return 0;
}
float GBamRecord::tag_float(const char tag[2]) { //get the float value of tag
uint8_t *s=bam_aux_get(this->b, tag);;
if (s) return ( bam_aux2f(s) );
return 0;
}
char* GBamRecord::tag_str(const char tag[2]) { //return string value for a tag
uint8_t *s=find_tag(tag);
if (s) return ( bam_aux2Z(s) );
return NULL;
}
char GBamRecord::spliceStrand() { // '+', '-' from the XS tag, or 0 if no XS tag
char c=tag_char1("XS");
if (c==0) {
//try minimap2's "ts" tag
char m=tag_char1("ts");
if (m=='+' || m=='-') {
if ((this->b->core.flag & BAM_FREVERSE) != 0) c=((m=='+') ? '-' : '+');
else c=m;
}
}
return ((c=='+' || c=='-') ? c : '.');
}
char* GBamRecord::sequence() { //user must free this after use
char *s = (char*)bam1_seq(b);
char* qseq=NULL;
GMALLOC(qseq, (b->core.l_qseq+1));
int i;
for (i=0;i<(b->core.l_qseq);i++) {
int8_t v = bam1_seqi(s,i);
qseq[i] = bam_nt16_rev_table[v];
}
qseq[i] = 0;
return qseq;
}
char* GBamRecord::qualities() {//user must free this after use
char *qual = (char*)bam1_qual(b);
char* qv=NULL;
GMALLOC(qv, (b->core.l_qseq+1) );
int i;
for(i=0;i<(b->core.l_qseq);i++) {
qv[i]=qual[i]+33;
}
qv[i]=0;
return qv;
}
char* GBamRecord::cigar() { //returns text version of the CIGAR string; must be freed by user
kstring_t str;
str.l = str.m = 0; str.s = 0;
if (b->core.n_cigar == 0) kputc('*', &str);
else {
for (int i = 0; i < b->core.n_cigar; ++i) {
kputw(bam1_cigar(b)[i]>>BAM_CIGAR_SHIFT, &str);
kputc("MIDNSHP=X"[bam1_cigar(b)[i]&BAM_CIGAR_MASK], &str);
}
}
return str.s;
}
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