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#include "samtools.pysam.h"
/* reset.c -- removes aligner updates and reference data from input sam /
bam / cram file and makes read data raw for new processing
Copyright (C) 2022 - 2024 Genome Research Ltd.
Author: Vasudeva Sarma <vasudeva.sarma@sanger.ac.uk>
Permission is hereby granted, free of charge, to any person obtaining a copy
of this software and associated documentation files (the "Software"), to deal
in the Software without restriction, including without limitation the rights
to use, copy, modify, merge, publish, distribute, sublicense, and/or sell
copies of the Software, and to permit persons to whom the Software is
furnished to do so, subject to the following conditions:
The above copyright notice and this permission notice shall be included in
all copies or substantial portions of the Software.
THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR
IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY,
FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL
THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER
LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING
FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER
DEALINGS IN THE SOFTWARE
*/
#include <config.h>
#include "samtools.h"
#include "htslib/sam.h"
#include "sam_opts.h"
#include "htslib/thread_pool.h"
#include "htslib/khash.h"
#include "sam_utils.h"
#include <unistd.h>
#define TAGNUM(X) (((X)[0] << 8) | (X)[1]) //to create key for aux tags, like type key in htslib
#define LONG_OPT(X) (128 + (X)) //to handle long and short options with same char
#define RESET_KEEPDUPFLAG 1 //keep dup flag as such, as in initial implementation
typedef struct conf_data
{
int keepRGs; //RG line handling
int noPGentry; //PG line for reset op or not
int ctrlFlags; //control flags
auxhash_t aux_keep; //SET that holds the aux tags to be retained
auxhash_t aux_remove; //SET that holds the aux tags to be removed
char *pgid; //PG id onwards which to be removed
} conf_data;
/// usage - print the usage
/** @param fp pointer to the file / terminal to which usage to be dumped
returns nothing
*/
static void usage(FILE *fp)
{
fprintf(fp, "Usage: samtools reset [options]\n\
-o FILE Output file\n\
-x, --remove-tag STR\n\
Aux tags to be removed\n\
--keep-tag STR\n\
Aux tags to be retained. Equivalent to -x ^STR\n\
--reject-PG ID\n\
Removes PG line with ID matching to input and succeeding PG lines\n\
--no-RG To have RG lines or not\n\
--no-PG To have PG entry or not for reset operation\n\
--dupflag\n\
Keeps the duplicate flag as it is\n");
sam_global_opt_help(fp, "--O--@--");
return;
}
/// update_aux_conf - update the user given aux tag configuration with defaults
/** @param config - pointer to conf_data
returns nothing
*/
void update_aux_conf(conf_data *config)
{
const char rg[] = "RG";
const char *default_tags[] = {"AS", "CC", "CG", "CP", "H1", "H2", "HI", "H0", "IH",
"MC", "MD", "MQ", "NM", "SA", "TS"};
khint_t iter = 0;
int ret = 0, i = 0;
if (!config)
return;
if (!config->aux_keep && !config->aux_remove) {
//none of aux tag filter in use, create remove filter
config->aux_remove = kh_init(aux_exists);
}
if (config->aux_keep) {
//keep set in use, remove RG if present
if (!config->keepRGs) {
iter = kh_get(aux_exists, config->aux_keep, TAGNUM(rg));
if (iter != kh_end(config->aux_keep)) {
kh_del(aux_exists, config->aux_keep, iter);
}
}
}
if (config->aux_remove) {
if (!config->keepRGs) {
//remove set in use, add RG if not present
iter = kh_get(aux_exists, config->aux_remove, TAGNUM(rg));
if (iter == kh_end(config->aux_remove)) {
kh_put(aux_exists, config->aux_remove, TAGNUM(rg), &ret);
}
}
//add the default tags if not present in remove set
//note, keep has priority and this may not be honoured
for (i = 0; i < sizeof(default_tags) / sizeof(default_tags[0]); ++i) {
iter = kh_get(aux_exists, config->aux_remove, TAGNUM(default_tags[i]));
if (iter == kh_end(config->aux_remove)) {
kh_put(aux_exists, config->aux_remove, TAGNUM(default_tags[i]), &ret);
}
}
}
}
/// removeauxtags - remove aux tags in bam data which are not present in acceptable tag set
/** @param bamdata - pointer to the bamdata from which needs the filtering
* @param config - pointer to conf_data
returns nothing
*/
void removeauxtags(bam1_t *bamdata, conf_data *config)
{
uint8_t *auxdata = NULL;
const char *tag = NULL;
khint_t iter = 0;
if (!bamdata || !config)
return;
for (auxdata = bam_aux_first(bamdata); auxdata; ) {
tag = bam_aux_tag(auxdata);
if (config->aux_keep) { //keep option or remove option with ^ in use
iter = kh_get(aux_exists, config->aux_keep, TAGNUM(tag));
if (iter == kh_end(config->aux_keep)) { //not present in keep, remove
auxdata = bam_aux_remove(bamdata, auxdata);
}
else { //present, keep
auxdata = bam_aux_next(bamdata, auxdata);
}
}
else if (config->aux_remove) { //remove option in use
iter = kh_get(aux_exists, config->aux_remove, TAGNUM(tag));
if (iter != kh_end(config->aux_remove)) { //present in remove, remove
auxdata = bam_aux_remove(bamdata, auxdata);
}
else { //not present, keep
auxdata = bam_aux_next(bamdata, auxdata);
}
}
//else impossible
}
}
/// getRGlines - add RG lines from input header to output header
/** @param in_samhdr - pointer to input sam header data
* @param out_samhdr - pointer to output sam header data
returns 1 on failure 0 on success
*/
int getRGlines(sam_hdr_t *in_samhdr, sam_hdr_t *out_samhdr)
{
kstring_t line = KS_INITIALIZE;
int i = 0, ret = 0, count = 0;
const char rg[] = "RG";
if (!in_samhdr || !out_samhdr) {
fprintf(samtools_stderr, "Invalid parameters in getRGlines!\n");
return 1;
}
if (-1 == (count = sam_hdr_count_lines(in_samhdr, rg))) {
fprintf(samtools_stderr, "Failed to get RG count!\n");
return 1;
}
for (i = 0; i < count; ++i)
{
ks_clear(&line);
if (sam_hdr_find_line_pos(in_samhdr, rg, i, &line)) {
fprintf(samtools_stderr, "Failed to get RG data!\n");
ret = 1;
break;
}
if (sam_hdr_add_lines(out_samhdr, line.s, line.l)) {
fprintf(samtools_stderr, "Failed to add RG data!\n");
ret = 1;
break;
}
}
ks_free(&line);
return ret;
}
/// getPGlines - add PG lines from input header to output header based on user option
/** @param in_samhdr - pointer to input sam header data
* @param out_samhdr - pointer to output sam header data
* @param config - pointer to internal configuration data
* @param argdump - string containing dump of command line invocation
returns 1 on failure 0 on success
*/
int getPGlines(sam_hdr_t *in_samhdr, sam_hdr_t *out_samhdr, conf_data *config, const char *argdump)
{
kstring_t line = KS_INITIALIZE, id = KS_INITIALIZE;
int i = 0, ret = 0, count = 0;
const char pg[] = "PG";
if (!in_samhdr || !out_samhdr || !config) {
fprintf(samtools_stderr, "Invalid parameters in getPGlines!\n");
return 1;
}
if (-1 == (count = sam_hdr_count_lines(in_samhdr, pg))) {
fprintf(samtools_stderr, "Failed to get PG count!\n");
return 1;
}
if (config->pgid && config->pgid[0]) { //when reject-PG is given, and is not empty, remove given pg onwards
for (i = 0; i < count; ++i) {
if (sam_hdr_find_tag_pos(in_samhdr, pg, i, "ID", &id)) {
fprintf(samtools_stderr, "Failed to get PG entry fields for line %d!\n", i + 1);
break;
}
if (!strcmp(id.s, config->pgid))
break;
//either current PG is prior to rejected one or all PGs are in, get PG line and add
ks_clear(&line);
if (sam_hdr_find_line_pos(in_samhdr, "PG", i, &line)) {
fprintf(samtools_stderr, "Failed to get PG data at %d!\n", i + 1);
ret = 1;
break;
}
//add to output
if (sam_hdr_add_lines(out_samhdr, line.s, line.l)) {
fprintf(samtools_stderr, "Failed to add PG data!\n");
ret = 1;
break;
}
}
}
else { //keep all
for (i = 0; i < count; ++i) {
if (sam_hdr_find_line_pos(in_samhdr, "PG", i, &line)) {
fprintf(samtools_stderr, "Failed to get PG data at %d!\n", i + 1);
ret = 1;
break;
}
//line has the required PG data
if (sam_hdr_add_lines(out_samhdr, line.s, line.l)) {
fprintf(samtools_stderr, "Failed to add PG data!\n");
ret = 1;
break;
}
}
}
if (!ret && !config->noPGentry) {
//add PG entry with reset command
if (-1 == (ret = sam_hdr_add_pg(out_samhdr, "samtools", "CL", argdump, NULL))) {
fprintf(samtools_stderr, "Failed to set PG entry!\n");
}
}
ks_free(&line);
ks_free(&id);
return ret;
}
/// reset - do the reset of data and create output; create output header with required rg/pg data, add bamdata with flags set to unmapped, pair info and orientation reset,
// reerse and complement alignment if required
/** @param infile - input samfile pointer
* @param outfile - output sam file pointer
* @param config - pointer to internal configuration data
* @param args - string containing dump of command line invocation
returns 1 on failure 0 on success
*/
int reset(samFile *infile, samFile *outfile, conf_data *config, char *args)
{
sam_hdr_t *in_samhdr = NULL, *out_samhdr = NULL;
int ret = EXIT_FAILURE, ret_r = 0, ret_w = 0, i = 0;
bam1_t *bamdata = NULL, *outdata = NULL;
kstring_t querydata = KS_INITIALIZE, qualdata = KS_INITIALIZE;
char *sp = NULL, *qp = NULL;
uint8_t *bamquery = NULL, *bamqual = NULL;
if (!infile || !outfile) {
fprintf(samtools_stderr, "Invalid parameters in reset!\n");
goto error;
}
//read input header
in_samhdr = sam_hdr_read(infile);
if (!in_samhdr)
{
fprintf(samtools_stderr, "Failed to read header from file!\n");
goto error;
}
//create output header
if (!(out_samhdr = sam_hdr_init()))
{
fprintf(samtools_stderr, "Failed to create output header!\n");
goto error;
}
//add version to output header
if (-1 == sam_hdr_add_line(out_samhdr,"HD", "VN", SAM_FORMAT_VERSION, NULL)) {
fprintf(samtools_stderr, "Failed to set header data!\n");
goto error;
}
//add RG / PG lines if configured
if ((config->keepRGs && getRGlines(in_samhdr, out_samhdr)) ||
getPGlines(in_samhdr, out_samhdr, config, args)) {
goto error;
}
//write output header
if (sam_hdr_write(outfile, out_samhdr)) {
print_error_errno("reset", "Output header write failed (%d)!\n", errno);
goto error;
}
bamdata = bam_init1(); //input bam
outdata = bam_init1(); //output bam
if (!bamdata || !outdata)
{
fprintf(samtools_stderr, "Failed to allocate data memory!\n");
goto error;
}
errno = 0; i = 0;
sp = NULL; qp = NULL;
bamquery = NULL; bamqual = NULL;
//get bam data, make updates and dump to output
while (0 <= (ret_r = sam_read1(infile, in_samhdr, bamdata)))
{
sp = NULL; qp = NULL;
bamquery = NULL; bamqual = NULL;
// read data
if (bamdata->core.flag & BAM_FSECONDARY || bamdata->core.flag & BAM_FSUPPLEMENTARY) {
continue;
}
//update flags
uint16_t flags = bamdata->core.flag & ~BAM_FPROPER_PAIR; //reset pair info
flags |= BAM_FUNMAP; //mark as unmapped
if (bamdata->core.flag & BAM_FPAIRED) {
flags |= BAM_FMUNMAP; //mark mate as unmapped, if it was a pair
}
flags &= ~BAM_FMREVERSE; //reset mate orientation
if (!(config->ctrlFlags & RESET_KEEPDUPFLAG)) {
flags &= ~BAM_FDUP; //reset dup flag from alignment
}
if (0 > ks_resize(&querydata, bamdata->core.l_qseq) ||
0 > ks_resize(&qualdata, bamdata->core.l_qseq)) {
fprintf(samtools_stderr, "Failed to get allocate memory!\n");
ret_r = -4;
break;
}
ks_clear(&querydata);
ks_clear(&qualdata);
sp = ks_str(&querydata);
qp = ks_str(&qualdata);
bamquery = bam_get_seq(bamdata);
bamqual = bam_get_qual(bamdata);
if (bamdata->core.flag & BAM_FREVERSE) {
//sequence data ordered as reverse complemented, reorder/complement sequence and quality data as read and clear the flag
for (i = bamdata->core.l_qseq - 1; i >= 0; --i) {
*sp++ = "=TGKCYSBAWRDMHVN"[bam_seqi(bamquery, i)];
*qp++ = bamqual[i];
}
flags &= ~BAM_FREVERSE; //reset flag as well
}
else {
//data in read order itself
for (i = 0; i < bamdata->core.l_qseq ; ++i) {
*sp++ = seq_nt16_str[bam_seqi(bamquery, i)];
}
memcpy(qp, bam_get_qual(bamdata), bamdata->core.l_qseq);
}
removeauxtags(bamdata, config);
if (0 > (ret_w = bam_set1(outdata, bamdata->core.l_qname - bamdata->core.l_extranul - 1, bam_get_qname(bamdata), flags, -1, -1, 0, 0, NULL, -1, -1, 0, bamdata->core.l_qseq, querydata.s, qualdata.s, bam_get_l_aux(bamdata)))) {
print_error_errno("reset", "Failed to set output data (%d)!\n", errno);
break;
}
memcpy(bam_get_aux(outdata), bam_get_aux(bamdata), bam_get_l_aux(bamdata));
outdata->l_data += bam_get_l_aux(bamdata);
errno = 0;
//write bam data to output
if (0 > (ret_w = sam_write1(outfile, out_samhdr, outdata)))
{
print_error_errno("reset", "Failed to write output data (%d)!\n", errno);
break;
}
// wrote the data, continue read/write cycle
errno = 0;
}
if (-1 > ret_r || 0 > ret_w) {
//some error
fprintf(samtools_stderr, "Error during %s!\n", (-1 > ret_r)? "read" : "write");
}
else {
// no error!
ret = EXIT_SUCCESS;
}
error:
// clean up and return result
if (in_samhdr)
sam_hdr_destroy(in_samhdr);
if (out_samhdr)
sam_hdr_destroy(out_samhdr);
if (bamdata)
bam_destroy1(bamdata);
if (outdata)
bam_destroy1(outdata);
if (qualdata.s)
ks_free(&qualdata);
if (querydata.s)
ks_free(&querydata);
return ret;
}
/// cleanup - free up allocations made
/** @param config - pointer to internal configuration data
returns nothing
*/
void cleanup(conf_data *config)
{
if (config->aux_keep) {
kh_destroy(aux_exists, config->aux_keep);
config->aux_keep = NULL;
}
if (config->aux_remove) {
kh_destroy(aux_exists, config->aux_remove);
config->aux_remove = NULL;
}
}
/// main_reset - starts the reset of data
/** @param argc - count of arguments
* @param argv - pointer to array of arguments
returns 1 on failure 0 on success
*/
int main_reset(int argc, char *argv[])
{
static const struct option lopts[] = {
SAM_OPT_GLOBAL_OPTIONS('-', '-', 'O', '-', '-', '@'), //let output format and thread count be given by user - long options
{"keep-tag", required_argument, NULL, LONG_OPT('x')}, //aux tags to be retained, supports ^ STR
{"remove-tag", required_argument, NULL, 'x'}, //aux tags to be removed
{"no-RG", no_argument, NULL, 1}, //no RG lines in output, default is to keep them
//reject PG lines from input, default is to keep them (i.e. option not given); without optional filename, all PGs removed and those given in file are filtered when optional filename is given
{"reject-PG", required_argument, NULL, 'p'}, //reject entries from this PG onwards
{"no-PG", no_argument, NULL, 2}, //do not add PG entry for reset operation, default is to add it
{"dupflag", no_argument, NULL, 3}, //keep the dup flag as it is - as in initial reset implementation
{NULL, 0, NULL, 0}
};
samFile *infile = NULL, *outfile = NULL;
sam_global_args ga = SAM_GLOBAL_ARGS_INIT;
htsThreadPool tpool = {NULL, 0};
const char *inname = NULL, *outname = NULL;
int c = 0, ret = EXIT_FAILURE;
char outmode[4] = "w", *args = NULL;
conf_data resetconf = {1, 0, 0, NULL, NULL, NULL}; //keep RGs and PGs by default, ctrlflags = 0
//samtools reset -o outfile -x/--remove-tag ... --keep-tag ... --threads=n --output-fmt=fmt --no-RG --reject-PG pgid --no-PG [<infile>]
while ((c = getopt_long(argc, argv, "o:@:x:O:", lopts, NULL)) >= 0)
{
switch (c)
{
case 1: //--no-RG
if (!resetconf.keepRGs) {
usage(samtools_stderr); //already given!
goto exit;
}
resetconf.keepRGs = 0;
break;
case 2: //--no-PG
if (resetconf.noPGentry) {
usage(samtools_stderr); //already given!
goto exit;
}
resetconf.noPGentry = 1;
break;
case 3: //keep dup flag as it is / no reset
resetconf.ctrlFlags |= RESET_KEEPDUPFLAG;
break;
case 'p': //--reject-PG=<id>
if (resetconf.pgid) {
usage(samtools_stderr); //already given!
goto exit;
}
resetconf.pgid = optarg;
break;
case 'o': //output file name
if (outname) { //already given!
usage(samtools_stderr);
goto exit;
}
outname = optarg;
break;
case 'x': //remove aux tag
if (*optarg == '^') { //remove all except given ones!
if (parse_aux_list(&resetconf.aux_keep, optarg+1, "main_reset")) {
usage(samtools_stderr);
goto exit;
}
}
else { //remove given ones
if (parse_aux_list(&resetconf.aux_remove, optarg, "main_reset")) {
usage(samtools_stderr);
goto exit;
}
}
break;
case LONG_OPT('x'): //keep aux tags
if (parse_aux_list(&resetconf.aux_keep, optarg, "main_reset")) {
usage(samtools_stderr);
goto exit;
}
break;
// handle standard samtool options like thread count, verbosity...
default:
if (parse_sam_global_opt(c, optarg, lopts, &ga) == 0)
break;
// else fall-through
// couldn't parse or unknown options, show usage!
case '?': //unknown options found!
usage(samtools_stderr);
goto exit;
break;
}
}
if (argc == 1 && isatty(STDIN_FILENO)) {
//no args and input is stdin -- it is the usage check
usage(samtools_stdout);
ret = EXIT_SUCCESS;
goto exit;
}
//else have other args or input from redirection/pipe/other device -- validate and work
if (!outname)
outname = "-";
//check and fail if unnecessary parameters are given
c = argc - optind;
if (c > 1) {
usage(samtools_stderr);
goto exit;
}
if (c == 1) {
inname = argv[optind];
}
else {
inname = "-";
}
//update aux tag configuration
update_aux_conf(&resetconf);
//set output file format based on name
sam_open_mode(outmode + 1, outname, NULL);
//open input and output files
infile = sam_open(inname, "r");
outfile = sam_open_format(outname, outmode, &ga.out);
if (!infile || !outfile) {
fprintf(samtools_stderr, "Could not open %s%s%s\n", !infile ? inname : "", (!infile && !outfile)? ", " : "", !outfile ? outname : "");
goto exit;
}
// set the thread count if given as argument
if (ga.nthreads > 0)
{
if (!(tpool.pool = hts_tpool_init(ga.nthreads)))
{
fprintf(samtools_stderr, "\nFailed to setup thread pool\n");
goto exit;
}
hts_set_opt(infile, HTS_OPT_THREAD_POOL, &tpool);
hts_set_opt(outfile, HTS_OPT_THREAD_POOL, &tpool);
}
args = stringify_argv(argc + 1, argv - 1); //to dump invocation in PG line
//do the reset!
ret = reset(infile, outfile, &resetconf, args);
exit:
if (args)
free(args);
if (infile)
sam_close(infile);
if (outfile)
sam_close(outfile);
if (tpool.pool)
hts_tpool_destroy(tpool.pool);
cleanup(&resetconf);
sam_global_args_free(&ga);
return ret;
}
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