int main(int argc, char **argv)
{
/* Global details */
AjBool explain_flag;
AjBool file_flag;
AjPStr* task;
AjBool do_primer;
AjBool do_hybrid;
ajint num_return;
ajint first_base_index;
/* "Sequence" Input Tags */
AjPSeqall sequence;
AjPRange included_region;
AjPRange target;
AjPRange excluded_region;
AjPStr left_input;
AjPStr right_input;
/* Primer details */
AjBool pick_anyway;
AjPFile mispriming_library;
float max_mispriming;
float pair_max_mispriming;
ajint gc_clamp;
ajint opt_size;
ajint min_size;
ajint max_size;
float opt_tm;
float min_tm;
float max_tm;
float max_diff_tm;
float opt_gc_percent;
float min_gc;
float max_gc;
float salt_conc;
float dna_conc;
ajint num_ns_accepted;
float self_any;
float self_end;
ajint max_poly_x;
/* Sequence Quality. These are not (yet) implemented */
/*
AjPFile sequence_quality;
ajint min_quality;
ajint min_end_quality;
ajint quality_range_min;
ajint quality_range_max;
*/
/* Product details */
ajint product_opt_size;
AjPRange product_size_range;
float product_opt_tm;
float product_min_tm;
float product_max_tm;
/* Objective Function Penalty Weights for Primers */
float max_end_stability;
/* these are not (yet) implemented */
/*
float inside_penalty;
float outside_penalty;
*/
/* Primer penalties */
/* these are not (yet) implemented */
/* Internal Oligo "Sequence" Input Tags */
AjPRange internal_oligo_excluded_region;
/* Internal Oligo "Global" Input Tags */
AjPStr internal_oligo_input;
ajint internal_oligo_opt_size;
ajint internal_oligo_min_size;
ajint internal_oligo_max_size;
float internal_oligo_opt_tm;
float internal_oligo_min_tm;
float internal_oligo_max_tm;
float internal_oligo_opt_gc_percent;
float internal_oligo_min_gc;
float internal_oligo_max_gc;
float internal_oligo_salt_conc;
float internal_oligo_dna_conc;
float internal_oligo_self_any;
float internal_oligo_self_end;
ajint internal_oligo_max_poly_x;
AjPFile internal_oligo_mishyb_library;
float internal_oligo_max_mishyb;
/*
ajint internal_oligo_min_quality;
*/
/* Internal Oligo penalties */
/* these are not (yet) implemented */
/* EMBOSS-wrapper-specific stuff */
AjPFile outfile;
/* other variables */
AjPStr result = NULL;
AjPStr strand = NULL;
AjPStr substr = NULL;
AjPSeq seq = NULL;
ajint begin = 0;
ajint end;
FILE* stream;
AjPStr taskstr = NULL;
const AjPStr program = NULL;
/* pipe variables */
int *pipeto; /* pipe to feed the exec'ed program input */
int *pipefrom; /* pipe to get the exec'ed program output */
embInit("eprimer3", argc, argv);
/* Global details */
explain_flag = ajAcdGetBoolean("explainflag");
file_flag = ajAcdGetBoolean("fileflag");
task = ajAcdGetList("task");
do_primer = ajAcdGetToggle("primer");
do_hybrid = ajAcdGetToggle("hybridprobe");
num_return = ajAcdGetInt("numreturn");
first_base_index = ajAcdGetInt("firstbaseindex");
/* "Sequence" Input Tags */
sequence = ajAcdGetSeqall("sequence");
included_region = ajAcdGetRange("includedregion");
target = ajAcdGetRange("targetregion");
excluded_region = ajAcdGetRange("excludedregion");
left_input = ajAcdGetString("forwardinput");
right_input = ajAcdGetString("reverseinput");
/* Primer details */
pick_anyway = ajAcdGetBoolean("pickanyway");
mispriming_library = ajAcdGetInfile("mispriminglibraryfile");
max_mispriming = ajAcdGetFloat("maxmispriming");
pair_max_mispriming = ajAcdGetFloat("pairmaxmispriming");
gc_clamp = ajAcdGetInt("gcclamp");
opt_size = ajAcdGetInt("osize");
min_size = ajAcdGetInt("minsize");
max_size = ajAcdGetInt("maxsize");
opt_tm = ajAcdGetFloat("otm");
min_tm = ajAcdGetFloat("mintm");
max_tm = ajAcdGetFloat("maxtm");
max_diff_tm = ajAcdGetFloat("maxdifftm");
opt_gc_percent = ajAcdGetFloat("ogcpercent");
min_gc = ajAcdGetFloat("mingc");
max_gc = ajAcdGetFloat("maxgc");
salt_conc = ajAcdGetFloat("saltconc");
dna_conc = ajAcdGetFloat("dnaconc");
num_ns_accepted = ajAcdGetInt("numnsaccepted");
self_any = ajAcdGetFloat("selfany");
self_end = ajAcdGetFloat("selfend");
max_poly_x = ajAcdGetInt("maxpolyx");
AJCNEW0(pipeto,2);
AJCNEW0(pipefrom,2);
/* Sequence Quality */
/* these are not (yet) implemented */
/*
sequence_quality = ajAcdGetInfile("sequencequality");
min_quality = ajAcdGetInt("minquality");
min_end_quality = ajAcdGetInt("minendquality");
quality_range_min = ajAcdGetInt("qualityrangemin");
quality_range_max = ajAcdGetInt("qualityrangemax");
*/
/* Product details */
product_opt_size = ajAcdGetInt("psizeopt");
product_size_range = ajAcdGetRange("prange");
product_opt_tm = ajAcdGetFloat("ptmopt");
product_min_tm = ajAcdGetFloat("ptmmin");
product_max_tm = ajAcdGetFloat("ptmmax");
/* Objective Function Penalty Weights for Primers */
max_end_stability = ajAcdGetFloat("maxendstability");
/* these are not (yet) implemented */
/*
inside_penalty = ajAcdGetFloat("insidepenalty");
outside_penalty = ajAcdGetFloat("outsidepenalty");
*/
/* Primer penalties */
/* these are not (yet) implemented */
/* Internal Oligo "Sequence" Input Tags */
internal_oligo_excluded_region = ajAcdGetRange("oexcludedregion");
internal_oligo_input = ajAcdGetString("oligoinput");
/* Internal Oligo "Global" Input Tags */
internal_oligo_opt_size = ajAcdGetInt("osizeopt");
internal_oligo_min_size = ajAcdGetInt("ominsize");
internal_oligo_max_size = ajAcdGetInt("omaxsize");
internal_oligo_opt_tm = ajAcdGetFloat("otmopt");
internal_oligo_min_tm = ajAcdGetFloat("otmmin");
internal_oligo_max_tm = ajAcdGetFloat("otmmax");
internal_oligo_opt_gc_percent = ajAcdGetFloat("ogcopt");
internal_oligo_min_gc = ajAcdGetFloat("ogcmin");
internal_oligo_max_gc = ajAcdGetFloat("ogcmax");
internal_oligo_salt_conc = ajAcdGetFloat("osaltconc");
internal_oligo_dna_conc = ajAcdGetFloat("odnaconc");
internal_oligo_self_any = ajAcdGetFloat("oanyself");
internal_oligo_self_end = ajAcdGetFloat("oendself");
internal_oligo_max_poly_x = ajAcdGetInt("opolyxmax");
internal_oligo_mishyb_library = ajAcdGetInfile("mishyblibraryfile");
internal_oligo_max_mishyb = ajAcdGetFloat("omishybmax");
/*
internal_oligo_min_quality = ajAcdGetInt("oligominquality");
*/
/* Internal Oligo penalties */
/* these are not (yet) implemented */
/* EMBOSS-wrapper-specific stuff */
outfile = ajAcdGetOutfile("outfile");
ajStrRemoveWhite(&left_input);
ajStrRemoveWhite(&right_input);
/*
** OK - we will now try to do a separate fork-exec for each sequence.
*/
result = ajStrNew();
while(ajSeqallNext(sequence, &seq))
{
program = ajAcdGetpathC("primer3_core");
if(!ajSysExecRedirectC(ajStrGetPtr(program),&pipeto,&pipefrom))
ajFatal("eprimer3: Could not exec primer3_core");
stream = eprimer3_start_write(pipeto[1]);
/* send primer3 Primer "Global" parameters */
eprimer3_send_bool(stream, "PRIMER_EXPLAIN_FLAG", explain_flag);
eprimer3_send_bool(stream, "PRIMER_FILE_FLAG", file_flag);
if(do_hybrid)
{
if(!ajStrCmpC(task[0], "1"))
ajStrAssignC(&taskstr, "pick_pcr_primers_and_hyb_probe");
else if(!ajStrCmpC(task[0], "2"))
ajStrAssignC(&taskstr, "pick_left_only");
else if(!ajStrCmpC(task[0], "3"))
ajStrAssignC(&taskstr, "pick_right_only");
else if(!ajStrCmpC(task[0], "4"))
ajStrAssignC(&taskstr, "pick_hyb_probe_only");
if (!do_primer)
ajStrAssignC(&taskstr, "pick_hyb_probe_only");
}
else
{
if(!ajStrCmpC(task[0], "1"))
ajStrAssignC(&taskstr, "pick_pcr_primers");
else if(!ajStrCmpC(task[0], "2"))
ajStrAssignC(&taskstr, "pick_left_only");
else if(!ajStrCmpC(task[0], "3"))
ajStrAssignC(&taskstr, "pick_right_only");
else if(!ajStrCmpC(task[0], "4"))
ajStrAssignC(&taskstr, "pick_hyb_probe_only");
}
eprimer3_send_string(stream, "PRIMER_TASK", taskstr);
eprimer3_send_int(stream, "PRIMER_NUM_RETURN", num_return);
eprimer3_send_int(stream, "PRIMER_FIRST_BASE_INDEX",
first_base_index);
eprimer3_send_bool(stream, "PRIMER_PICK_ANYWAY", pick_anyway);
/* mispriming library may not have been specified */
if(mispriming_library)
eprimer3_send_stringC(stream, "PRIMER_MISPRIMING_LIBRARY",
ajFileGetPrintnameC(mispriming_library));
eprimer3_send_float(stream, "PRIMER_MAX_MISPRIMING",
max_mispriming);
eprimer3_send_float(stream, "PRIMER_PAIR_MAX_MISPRIMING",
pair_max_mispriming);
eprimer3_send_int(stream, "PRIMER_GC_CLAMP", gc_clamp);
eprimer3_send_int(stream, "PRIMER_OPT_SIZE", opt_size);
eprimer3_send_int(stream, "PRIMER_MIN_SIZE", min_size);
eprimer3_send_int(stream, "PRIMER_MAX_SIZE", max_size);
eprimer3_send_float(stream, "PRIMER_OPT_TM", opt_tm);
eprimer3_send_float(stream, "PRIMER_MIN_TM", min_tm);
eprimer3_send_float(stream, "PRIMER_MAX_TM", max_tm);
eprimer3_send_float(stream, "PRIMER_MAX_DIFF_TM", max_diff_tm);
eprimer3_send_float(stream, "PRIMER_OPT_GC_PERCENT",
opt_gc_percent);
eprimer3_send_float(stream, "PRIMER_MIN_GC", min_gc);
eprimer3_send_float(stream, "PRIMER_MAX_GC", max_gc);
eprimer3_send_float(stream, "PRIMER_SALT_CONC", salt_conc);
eprimer3_send_float(stream, "PRIMER_DNA_CONC", dna_conc);
eprimer3_send_int(stream, "PRIMER_NUM_NS_ACCEPTED",
num_ns_accepted);
eprimer3_send_float(stream, "PRIMER_SELF_ANY", self_any);
eprimer3_send_float(stream, "PRIMER_SELF_END", self_end);
eprimer3_send_int(stream, "PRIMER_MAX_POLY_X", max_poly_x);
eprimer3_send_int(stream, "PRIMER_PRODUCT_OPT_SIZE",
product_opt_size);
eprimer3_send_range2(stream, "PRIMER_PRODUCT_SIZE_RANGE",
product_size_range);
eprimer3_send_float(stream, "PRIMER_PRODUCT_OPT_TM",
product_opt_tm);
eprimer3_send_float(stream, "PRIMER_PRODUCT_MIN_TM",
product_min_tm);
eprimer3_send_float(stream, "PRIMER_PRODUCT_MAX_TM",
product_max_tm);
eprimer3_send_float(stream, "PRIMER_MAX_END_STABILITY",
max_end_stability);
/* send primer3 Internal Oligo "Global" parameters */
eprimer3_send_int(stream, "PRIMER_INTERNAL_OLIGO_OPT_SIZE",
internal_oligo_opt_size);
eprimer3_send_int(stream, "PRIMER_INTERNAL_OLIGO_MIN_SIZE",
internal_oligo_min_size);
eprimer3_send_int(stream, "PRIMER_INTERNAL_OLIGO_MAX_SIZE",
internal_oligo_max_size);
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_OPT_TM",
internal_oligo_opt_tm);
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_MIN_TM",
internal_oligo_min_tm);
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_MAX_TM",
internal_oligo_max_tm);
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_OPT_GC_PERCENT",
internal_oligo_opt_gc_percent);
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_MIN_GC",
internal_oligo_min_gc);
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_MAX_GC",
internal_oligo_max_gc);
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_SALT_CONC",
internal_oligo_salt_conc);
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_DNA_CONC",
internal_oligo_dna_conc);
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_SELF_ANY",
internal_oligo_self_any);
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_SELF_END",
internal_oligo_self_end);
eprimer3_send_int(stream, "PRIMER_INTERNAL_OLIGO_MAX_POLY_X",
internal_oligo_max_poly_x);
/*
** internal oligo mishybridising library may not have been
** specified
*/
if(internal_oligo_mishyb_library)
eprimer3_send_stringC(stream,
"PRIMER_INTERNAL_OLIGO_MISHYB_LIBRARY",
ajFileGetPrintnameC(internal_oligo_mishyb_library));
eprimer3_send_float(stream, "PRIMER_INTERNAL_OLIGO_MAX_MISHYB",
internal_oligo_max_mishyb);
/*
** Start sequence-specific stuff
*/
begin = ajSeqallGetseqBegin(sequence) - 1;
end = ajSeqallGetseqEnd(sequence) - 1;
strand = ajSeqGetSeqCopyS(seq);
ajStrFmtUpper(&strand);
ajStrAssignSubC(&substr,ajStrGetPtr(strand), begin, end);
/* send flags to turn on using optimal product size */
eprimer3_send_float(stream, "PRIMER_PAIR_WT_PRODUCT_SIZE_GT",
(float)0.05);
eprimer3_send_float(stream, "PRIMER_PAIR_WT_PRODUCT_SIZE_LT",
(float)0.05);
/* send primer3 Primer "Sequence" parameters */
eprimer3_send_string(stream, "SEQUENCE", substr);
/* if no ID name, use the USA */
if(ajStrMatchC(ajSeqGetNameS(seq),""))
eprimer3_send_string(stream, "PRIMER_SEQUENCE_ID",
ajSeqGetUsaS(seq));
else
eprimer3_send_string(stream, "PRIMER_SEQUENCE_ID",
ajSeqGetNameS(seq));
eprimer3_send_range(stream, "INCLUDED_REGION", included_region,
begin);
eprimer3_send_range(stream, "TARGET", target, begin);
eprimer3_send_range(stream, "EXCLUDED_REGION", excluded_region,
begin);
eprimer3_send_string(stream, "PRIMER_LEFT_INPUT", left_input);
eprimer3_send_string(stream, "PRIMER_RIGHT_INPUT", right_input);
/* send primer3 Internal Oligo "Sequence" parameters */
eprimer3_send_range(stream,
"PRIMER_INTERNAL_OLIGO_EXCLUDED_REGION",
internal_oligo_excluded_region, begin);
eprimer3_send_string(stream, "PRIMER_INTERNAL_OLIGO_INPUT",
internal_oligo_input);
/* end the primer3 input sequence record with a '=' */
eprimer3_send_end(stream);
/* and close the ouput pipe stream */
eprimer3_end_write(stream);
/* read the primer3 output */
eprimer3_read(pipefrom[0], &result);
eprimer3_report(outfile, result, num_return, begin);
ajStrSetClear(&result);
#ifndef WIN32
close(pipeto[1]);
close(pipefrom[0]);
#endif
} /* end of sequence loop */
ajStrDel(&result);
ajSeqDel(&seq);
ajStrDel(&strand);
ajStrDel(&substr);
ajFileClose(&outfile);
ajStrDel(&taskstr);
ajStrDelarray(&task);
ajSeqallDel(&sequence);
ajSeqDel(&seq);
ajRangeDel(&included_region);
ajRangeDel(&target);
ajRangeDel(&excluded_region);
ajRangeDel(&product_size_range);
ajRangeDel(&internal_oligo_excluded_region);
ajStrDel(&left_input);
ajStrDel(&right_input);
ajStrDel(&internal_oligo_input);
AJFREE(pipeto);
AJFREE(pipefrom);
ajFileClose(&mispriming_library);
embExit();
return 0;
}
int main(int argc, char **argv)
{
AjPSeqout outseq = NULL;
AjPList list = NULL;
AjPSeq seq = NULL;
AjPStr insert = NULL;
AjPStr seqstr = NULL;
AjPStr* seqr = NULL;
AjPFile data = NULL;
ajint start = 0;
ajint length = 0;
ajint amount = 0;
ajint scmax = 0;
ajint extra = 0;
embInit("makeprotseq", argc, argv);
data = ajAcdGetInfile("pepstatsfile");
insert = ajAcdGetString("insert");
start = ajAcdGetInt("start");
length = ajAcdGetInt("length");
amount = ajAcdGetInt("amount");
outseq = ajAcdGetSeqoutall("outseq");
list = ajListstrNew();
/* this is checked by acd
if(amount <=0 || length <= 0)
ajFatal("Amount or length is 0 or less. "
"Unable to create any sequences"); */
/* if insert, make sure sequence is large enough */
if(ajStrGetLen(insert))
{
length -= ajStrGetLen(insert);
/* start= start <= 1 ? 0 : --start; */ /* checked in acd */
start--;
if(length <= 0)
ajFatal("Sequence smaller than inserted part. "
"Unable to create sequences.");
}
/* make the list of AjPStr to be used in sequence creation */
if(data)
{
ajDebug("Distribution datafile '%s' given checking type\n",
ajFileGetPrintnameC(data));
seqstr = ajStrNew();
ajReadlineTrim(data,&seqstr);
if(ajStrFindC(seqstr,"PEPSTATS") == 0)
{
makeprotseq_parse_pepstats(&list,data);
}
else
{
ajWarn("Not pepstats file. Making completely random sequences.");
makeprotseq_default_chars(&list);
}
ajStrDel(&seqstr);
ajFileClose(&data);
}
else
makeprotseq_default_chars(&list);
/* if insert, make sure type is correct */
/* typecheking code is not working, uncomment and test after it is
if(ajStrGetLen(insert))
{
seqstr = ajStrNew();
if(prot)
ajStrAssignC(&seqstr,"pureprotein");
if(!ajSeqTypeCheckS(&insert,seqstr))
ajFatal("Insert not the same sequence type as sequence itself.");
ajStrDel(&seqstr);
} */
/* array allows fast creation of a sequences */
scmax = (ajuint) ajListstrToarray(list,&seqr);
if(!scmax)
ajFatal("No strings in list. No characters to make the sequence.");
ajDebug("Distribution array done.\nscmax '%d', extra '%d', first '%S'\n",
scmax,extra,seqr[0]);
ajRandomSeed();
while(amount-- > 0)
{
seqstr = makeprotseq_random_sequence(seqr,scmax,length);
if(ajStrGetLen(insert))
ajStrInsertS(&seqstr,start,insert);
ajStrFmtLower(&seqstr);
seq = ajSeqNew();
ajSeqAssignSeqS(seq, seqstr);
ajSeqSetProt(seq);
ajSeqoutWriteSeq(outseq, seq);
ajSeqDel(&seq);
ajStrDel(&seqstr);
}
ajSeqoutClose(outseq);
ajSeqoutDel(&outseq);
ajListstrFreeData(&list);
ajStrDel(&insert);
AJFREE(seqr);
embExit();
return 0;
}
