int main(int argc, char *argv[]){
struct RNA2Dfold_args_info args_info;
unsigned int input_type;
char *string, *input_string, *orig_sequence;
char *mfe_structure=NULL, *structure1=NULL, *structure2=NULL, *reference_struc1=NULL, *reference_struc2=NULL;
char *ParamFile=NULL;
int i, j, length, l;
double min_en;
double kT, sfact=1.07;
int pf=0,istty;
int noconv=0;
int circ=0;
int maxDistance1 = -1;
int maxDistance2 = -1;
int do_backtrack = 1;
int stBT = 0;
int nstBT = 0;
string=NULL;
dangles = 2;
struct nbhoods *neighborhoods = NULL;
struct nbhoods *neighborhoods_cur = NULL;
string = input_string = orig_sequence = NULL;
/*
#############################################
# check the command line prameters
#############################################
*/
if(RNA2Dfold_cmdline_parser (argc, argv, &args_info) != 0) exit(1);
/* temperature */
if(args_info.temp_given)
temperature = args_info.temp_arg;
/* max distance to 1st reference structure */
if(args_info.maxDist1_given)
maxDistance1 = args_info.maxDist1_arg;
/* max distance to 2nd reference structure */
if(args_info.maxDist2_given)
maxDistance2 = args_info.maxDist2_arg;
/* compute partition function and boltzmann probabilities */
if(args_info.partfunc_given)
pf = 1;
/* do stachastic backtracking */
if(args_info.stochBT_given){
pf = 1;
stBT = 1;
nstBT = args_info.stochBT_arg;
}
if(args_info.noTetra_given)
tetra_loop=0;
/* assume RNA sequence to be circular */
if(args_info.circ_given)
circ=1;
/* dangle options */
if(args_info.dangles_given)
dangles=args_info.dangles_arg;
/* set number of threads for parallel computation */
if(args_info.numThreads_given)
#ifdef _OPENMP
omp_set_num_threads(args_info.numThreads_arg);
#else
nrerror("\'j\' option is available only if compiled with OpenMP support!");
#endif
/* get energy parameter file name */
if(args_info.parameterFile_given)
ParamFile = strdup(args_info.parameterFile_arg);
/* do not allow GU pairs ? */
if(args_info.noGU_given)
noGU = 1;
/* do not allow GU pairs at the end of helices? */
if(args_info.noClosingGU_given)
no_closingGU = 1;
/* pf scaling factor */
if(args_info.pfScale_given)
sfact = args_info.pfScale_arg;
/* do not backtrack structures ? */
if(args_info.noBT_given)
do_backtrack = 0;
for (i = 0; i < args_info.neighborhood_given; i++){
int kappa, lambda;
kappa = lambda = 0;
if(sscanf(args_info.neighborhood_arg[i], "%d:%d", &kappa, &lambda) == 2);
if ((kappa>-2) && (lambda>-2)){
if(neighborhoods_cur != NULL){
neighborhoods_cur->next = (nbhoods *)space(sizeof(nbhoods));
neighborhoods_cur = neighborhoods_cur->next;
}
else{
neighborhoods = (nbhoods *)space(sizeof(nbhoods));
neighborhoods_cur = neighborhoods;
}
neighborhoods_cur->k = kappa;
neighborhoods_cur->l = lambda;
neighborhoods_cur->next = NULL;
}
}
/* free allocated memory of command line data structure */
RNA2Dfold_cmdline_parser_free (&args_info);
/*
#############################################
# begin actual program code
#############################################
*/
if (ParamFile != NULL)
read_parameter_file(ParamFile);
istty = isatty(fileno(stdout))&&isatty(fileno(stdin));
/*
#############################################
# main loop, continue until end of file
#############################################
*/
do {
if (istty)
print_tty_input_seq_str("Input strings\n1st line: sequence (upper or lower case)\n2nd + 3rd line: reference structures (dot bracket notation)\n@ to quit\n");
while((input_type = get_input_line(&input_string, 0)) & VRNA_INPUT_FASTA_HEADER){
printf(">%s\n", input_string); /* print fasta header if available */
free(input_string);
}
/* break on any error, EOF or quit request */
if(input_type & (VRNA_INPUT_QUIT | VRNA_INPUT_ERROR)){ break;}
/* else assume a proper sequence of letters of a certain alphabet (RNA, DNA, etc.) */
else{
length = (int) strlen(input_string);
string = strdup(input_string);
free(input_string);
}
mfe_structure = (char *) space((unsigned) length+1);
structure1 = (char *) space((unsigned) length+1);
structure2 = (char *) space((unsigned) length+1);
input_type = get_input_line(&input_string, VRNA_INPUT_NOSKIP_COMMENTS);
if(input_type & VRNA_INPUT_QUIT){ break;}
else if((input_type & VRNA_INPUT_MISC) && (strlen(input_string) > 0)){
reference_struc1 = strdup(input_string);
free(input_string);
if(strlen(reference_struc1) != length)
nrerror("sequence and 1st reference structure have unequal length");
}
else nrerror("1st reference structure missing\n");
strncpy(structure1, reference_struc1, length);
input_type = get_input_line(&input_string, VRNA_INPUT_NOSKIP_COMMENTS);
if(input_type & VRNA_INPUT_QUIT){ break;}
else if((input_type & VRNA_INPUT_MISC) && (strlen(input_string) > 0)){
reference_struc2 = strdup(input_string);
free(input_string);
if(strlen(reference_struc2) != length)
nrerror("sequence and 2nd reference structure have unequal length");
}
else nrerror("2nd reference structure missing\n");
strncpy(structure2, reference_struc2, length);
/* convert DNA alphabet to RNA if not explicitely switched off */
if(!noconv) str_DNA2RNA(string);
/* store case-unmodified sequence */
orig_sequence = strdup(string);
/* convert sequence to uppercase letters only */
str_uppercase(string);
if (istty) printf("length = %d\n", length);
min_en = (circ) ? circfold(string, mfe_structure) : fold(string, mfe_structure);
printf("%s\n%s", orig_sequence, mfe_structure);
if (istty)
printf("\n minimum free energy = %6.2f kcal/mol\n", min_en);
else
printf(" (%6.2f)\n", min_en);
printf("%s (%6.2f) <ref 1>\n", structure1, (circ) ? energy_of_circ_structure(string, structure1, 0) : energy_of_structure(string,structure1, 0));
printf("%s (%6.2f) <ref 2>\n", structure2, (circ) ? energy_of_circ_structure(string, structure2, 0) : energy_of_structure(string,structure2, 0));
/* get all variables need for the folding process (some memory will be preallocated here too) */
TwoDfold_vars *mfe_vars = get_TwoDfold_variables(string, structure1, structure2, circ);
mfe_vars->do_backtrack = do_backtrack;
TwoDfold_solution *mfe_s = TwoDfoldList(mfe_vars, maxDistance1, maxDistance2);
if(!pf){
#ifdef COUNT_STATES
printf("k\tl\tn\tMFE\tMFE-structure\n");
for(i = 0; mfe_s[i].k != INF; i++){
printf("%d\t%d\t%lu\t%6.2f\t%s\n", mfe_s[i].k, mfe_s[i].l, mfe_vars->N_F5[length][mfe_s[i].k][mfe_s[i].l/2], mfe_s[i].en, mfe_s[i].s);
if(mfe_s[i].s) free(mfe_s[i].s);
}
free(mfe_s);
#else
printf("k\tl\tMFE\tMFE-structure\n");
for(i = 0; mfe_s[i].k != INF; i++){
printf("%d\t%d\t%6.2f\t%s\n", mfe_s[i].k, mfe_s[i].l, mfe_s[i].en, mfe_s[i].s);
if(mfe_s[i].s) free(mfe_s[i].s);
}
free(mfe_s);
#endif
}
if(pf){
int maxD1 = (int) mfe_vars->maxD1;
int maxD2 = (int) mfe_vars->maxD2;
float mmfe = INF;
double Q;
for(i = 0; mfe_s[i].k != INF; i++){
if(mmfe > mfe_s[i].en) mmfe = mfe_s[i].en;
}
kT = (temperature+K0)*GASCONST/1000.0; /* in Kcal */
pf_scale = exp(-(sfact*mmfe)/kT/length);
if (length>2000)
fprintf(stdout, "scaling factor %f\n", pf_scale);
/* get all variables need for the folding process (some memory will be preallocated there too) */
//TwoDpfold_vars *q_vars = get_TwoDpfold_variables_from_MFE(mfe_vars);
/* we dont need the mfe vars and arrays anymore, so we can savely free their occupying memory */
destroy_TwoDfold_variables(mfe_vars);
TwoDpfold_vars *q_vars = get_TwoDpfold_variables(string, structure1, structure2, circ);
TwoDpfold_solution *pf_s = TwoDpfoldList(q_vars, maxD1, maxD2);
Q = 0.;
for(i = 0; pf_s[i].k != INF; i++){
Q += pf_s[i].q;
}
double fee = (-log(Q)-length*log(pf_scale))*kT;
if(!stBT){
printf("free energy of ensemble = %6.2f kcal/mol\n",fee);
printf("k\tl\tP(neighborhood)\tP(MFE in neighborhood)\tP(MFE in ensemble)\tMFE\tE_gibbs\tMFE-structure\n");
for(i=0; pf_s[i].k != INF;i++){
float free_energy = (-log((float)pf_s[i].q)-length*log(pf_scale))*kT;
if((pf_s[i].k != mfe_s[i].k) || (pf_s[i].l != mfe_s[i].l))
nrerror("This should never happen!");
fprintf(stdout,
"%d\t%d\t%2.8f\t%2.8f\t%2.8f\t%6.2f\t%6.2f\t%s\n",
pf_s[i].k,
pf_s[i].l,
(float)(pf_s[i].q)/(float)Q,
exp((free_energy-mfe_s[i].en)/kT),
exp((fee-mfe_s[i].en)/kT),
mfe_s[i].en,
free_energy,
mfe_s[i].s);
}
}
else{
init_rand();
if(neighborhoods != NULL){
nbhoods *tmp, *tmp2;
for(tmp = neighborhoods; tmp != NULL; tmp = tmp->next){
int k,l;
k = tmp->k;
l = tmp->l;
for(i = 0; i < nstBT; i++){
char *s = TwoDpfold_pbacktrack(q_vars, k, l);
printf("%d\t%d\t%s\t%6.2f\n", k, l, s, q_vars->circ ? energy_of_circ_structure(q_vars->sequence, s, 0) : energy_of_structure(q_vars->sequence, s, 0));
}
}
}
else{
for(i=0; pf_s[i].k != INF;i++){
for(l = 0; l < nstBT; l++){
char *s = TwoDpfold_pbacktrack(q_vars, pf_s[i].k, pf_s[i].l);
printf("%d\t%d\t%s\t%6.2f\n", pf_s[i].k, pf_s[i].l, s, q_vars->circ ? energy_of_circ_structure(q_vars->sequence, s, 0) : energy_of_structure(q_vars->sequence, s, 0));
}
}
}
}
free_pf_arrays();
for(i=0; mfe_s[i].k != INF;i++){
if(mfe_s[i].s) free(mfe_s[i].s);
}
free(pf_s);
free(mfe_s);
/* destroy the q_vars */
destroy_TwoDpfold_variables(q_vars);
}
else
destroy_TwoDfold_variables(mfe_vars);
free_arrays();
free(string);
free(orig_sequence);
free(mfe_structure);
free(structure1);
free(structure2);
free(reference_struc1);
free(reference_struc2);
string = orig_sequence = mfe_structure = NULL;
} while (1);
return 0;
}
int main(int argc, char *argv[])
{
struct RNAcofold_args_info args_info;
unsigned int input_type;
char *string, *input_string;
char *structure, *cstruc, *rec_sequence, *orig_sequence, *rec_id, **rec_rest;
char fname[FILENAME_MAX_LENGTH], ffname[FILENAME_MAX_LENGTH];
char *ParamFile;
char *ns_bases, *c;
char *Concfile;
int i, length, l, sym, r, cl;
double min_en;
double kT, sfact, betaScale;
int pf, istty;
int noconv, noPS;
int doT; /*compute dimere free energies etc.*/
int doC; /*toggle to compute concentrations*/
int doQ; /*toggle to compute prob of base being paired*/
int cofi; /*toggle concentrations stdin / file*/
plist *prAB;
plist *prAA; /*pair probabilities of AA dimer*/
plist *prBB;
plist *prA;
plist *prB;
plist *mfAB;
plist *mfAA; /*pair mfobabilities of AA dimer*/
plist *mfBB;
plist *mfA;
plist *mfB;
double *ConcAandB;
unsigned int rec_type, read_opt;
pf_paramT *pf_parameters;
model_detailsT md;
/*
#############################################
# init variables and parameter options
#############################################
*/
dangles = 2;
sfact = 1.07;
bppmThreshold = 1e-5;
noconv = 0;
noPS = 0;
do_backtrack = 1;
pf = 0;
doT = 0;
doC = 0;
doQ = 0;
cofi = 0;
betaScale = 1.;
gquad = 0;
ParamFile = NULL;
pf_parameters = NULL;
string = NULL;
Concfile = NULL;
structure = NULL;
cstruc = NULL;
ns_bases = NULL;
rec_type = read_opt = 0;
rec_id = rec_sequence = orig_sequence = NULL;
rec_rest = NULL;
set_model_details(&md);
/*
#############################################
# check the command line prameters
#############################################
*/
if(RNAcofold_cmdline_parser (argc, argv, &args_info) != 0) exit(1);
/* temperature */
if(args_info.temp_given) temperature = args_info.temp_arg;
/* structure constraint */
if(args_info.constraint_given) fold_constrained=1;
/* do not take special tetra loop energies into account */
if(args_info.noTetra_given) md.special_hp = tetra_loop=0;
/* set dangle model */
if(args_info.dangles_given){
if((args_info.dangles_arg < 0) || (args_info.dangles_arg > 3))
warn_user("required dangle model not implemented, falling back to default dangles=2");
else
md.dangles = dangles = args_info.dangles_arg;
}
/* do not allow weak pairs */
if(args_info.noLP_given) md.noLP = noLonelyPairs = 1;
/* do not allow wobble pairs (GU) */
if(args_info.noGU_given) md.noGU = noGU = 1;
/* do not allow weak closing pairs (AU,GU) */
if(args_info.noClosingGU_given) md.noGUclosure = no_closingGU = 1;
/* gquadruplex support */
if(args_info.gquad_given) md.gquad = gquad = 1;
/* enforce canonical base pairs in any case? */
if(args_info.canonicalBPonly_given) md.canonicalBPonly = canonicalBPonly = 1;
/* do not convert DNA nucleotide "T" to appropriate RNA "U" */
if(args_info.noconv_given) noconv = 1;
/* set energy model */
if(args_info.energyModel_given) energy_set = args_info.energyModel_arg;
/* */
if(args_info.noPS_given) noPS = 1;
/* take another energy parameter set */
if(args_info.paramFile_given) ParamFile = strdup(args_info.paramFile_arg);
/* Allow other pairs in addition to the usual AU,GC,and GU pairs */
if(args_info.nsp_given) ns_bases = strdup(args_info.nsp_arg);
/* set pf scaling factor */
if(args_info.pfScale_given) sfact = args_info.pfScale_arg;
if(args_info.all_pf_given) doT = pf = 1;
/* concentrations from stdin */
if(args_info.concentrations_given) doC = doT = pf = 1;
/* set the bppm threshold for the dotplot */
if(args_info.bppmThreshold_given)
bppmThreshold = MIN2(1., MAX2(0.,args_info.bppmThreshold_arg));
/* concentrations in file */
if(args_info.betaScale_given) betaScale = args_info.betaScale_arg;
if(args_info.concfile_given){
Concfile = strdup(args_info.concfile_arg);
doC = cofi = doT = pf = 1;
}
/* partition function settings */
if(args_info.partfunc_given){
pf = 1;
if(args_info.partfunc_arg != -1)
do_backtrack = args_info.partfunc_arg;
}
/* free allocated memory of command line data structure */
RNAcofold_cmdline_parser_free (&args_info);
/*
#############################################
# begin initializing
#############################################
*/
if(pf && gquad){
nrerror("G-Quadruplex support is currently not available for partition function computations");
}
if (ParamFile != NULL)
read_parameter_file(ParamFile);
if (ns_bases != NULL) {
nonstandards = space(33);
c=ns_bases;
i=sym=0;
if (*c=='-') {
sym=1; c++;
}
while (*c!='\0') {
if (*c!=',') {
nonstandards[i++]=*c++;
nonstandards[i++]=*c;
if ((sym)&&(*c!=*(c-1))) {
nonstandards[i++]=*c;
nonstandards[i++]=*(c-1);
}
}
c++;
}
}
istty = isatty(fileno(stdout))&&isatty(fileno(stdin));
/* print user help if we get input from tty */
if(istty){
printf("Use '&' to connect 2 sequences that shall form a complex.\n");
if(fold_constrained){
print_tty_constraint(VRNA_CONSTRAINT_DOT | VRNA_CONSTRAINT_X | VRNA_CONSTRAINT_ANG_BRACK | VRNA_CONSTRAINT_RND_BRACK);
print_tty_input_seq_str("Input sequence (upper or lower case) followed by structure constraint\n");
}
else print_tty_input_seq();
}
/* set options we wanna pass to read_record */
if(istty) read_opt |= VRNA_INPUT_NOSKIP_BLANK_LINES;
if(!fold_constrained) read_opt |= VRNA_INPUT_NO_REST;
/*
#############################################
# main loop: continue until end of file
#############################################
*/
while(
!((rec_type = read_record(&rec_id, &rec_sequence, &rec_rest, read_opt))
& (VRNA_INPUT_ERROR | VRNA_INPUT_QUIT))){
/*
########################################################
# init everything according to the data we've read
########################################################
*/
if(rec_id){
if(!istty) printf("%s\n", rec_id);
(void) sscanf(rec_id, ">%" XSTR(FILENAME_ID_LENGTH) "s", fname);
}
else fname[0] = '\0';
cut_point = -1;
rec_sequence = tokenize(rec_sequence); /* frees input_string and sets cut_point */
length = (int) strlen(rec_sequence);
structure = (char *) space((unsigned) length+1);
/* parse the rest of the current dataset to obtain a structure constraint */
if(fold_constrained){
cstruc = NULL;
int cp = cut_point;
unsigned int coptions = (rec_id) ? VRNA_CONSTRAINT_MULTILINE : 0;
coptions |= VRNA_CONSTRAINT_DOT | VRNA_CONSTRAINT_X | VRNA_CONSTRAINT_ANG_BRACK | VRNA_CONSTRAINT_RND_BRACK;
getConstraint(&cstruc, (const char **)rec_rest, coptions);
cstruc = tokenize(cstruc);
if(cut_point != cp) nrerror("cut point in sequence and structure constraint differs");
cl = (cstruc) ? (int)strlen(cstruc) : 0;
if(cl == 0) warn_user("structure constraint is missing");
else if(cl < length) warn_user("structure constraint is shorter than sequence");
else if(cl > length) nrerror("structure constraint is too long");
if(cstruc) strncpy(structure, cstruc, sizeof(char)*(cl+1));
}
/* convert DNA alphabet to RNA if not explicitely switched off */
if(!noconv) str_DNA2RNA(rec_sequence);
/* store case-unmodified sequence */
orig_sequence = strdup(rec_sequence);
/* convert sequence to uppercase letters only */
str_uppercase(rec_sequence);
if(istty){
if (cut_point == -1)
printf("length = %d\n", length);
else
printf("length1 = %d\nlength2 = %d\n", cut_point-1, length-cut_point+1);
}
/*
########################################################
# begin actual computations
########################################################
*/
if (doC) {
FILE *fp;
if (cofi) { /* read from file */
fp = fopen(Concfile, "r");
if (fp==NULL) {
fprintf(stderr, "could not open concentration file %s", Concfile);
nrerror("\n");
}
ConcAandB = read_concentrations(fp);
fclose(fp);
} else {
printf("Please enter concentrations [mol/l]\n format: ConcA ConcB\n return to end\n");
ConcAandB = read_concentrations(stdin);
}
}
/*compute mfe of AB dimer*/
min_en = cofold(rec_sequence, structure);
assign_plist_from_db(&mfAB, structure, 0.95);
{
char *pstring, *pstruct;
if (cut_point == -1) {
pstring = strdup(orig_sequence);
pstruct = strdup(structure);
} else {
pstring = costring(orig_sequence);
pstruct = costring(structure);
}
printf("%s\n%s", pstring, pstruct);
if (istty)
printf("\n minimum free energy = %6.2f kcal/mol\n", min_en);
else
printf(" (%6.2f)\n", min_en);
(void) fflush(stdout);
if (!noPS) {
char annot[512] = "";
if (fname[0]!='\0') {
strcpy(ffname, fname);
strcat(ffname, "_ss.ps");
} else {
strcpy(ffname, "rna.ps");
}
if (cut_point >= 0)
sprintf(annot,
"1 %d 9 0 0.9 0.2 omark\n%d %d 9 1 0.1 0.2 omark\n",
cut_point-1, cut_point+1, length+1);
if(gquad){
if (!noPS) (void) PS_rna_plot_a_gquad(pstring, pstruct, ffname, annot, NULL);
} else {
if (!noPS) (void) PS_rna_plot_a(pstring, pstruct, ffname, annot, NULL);
}
}
free(pstring);
free(pstruct);
}
if (length>2000) free_co_arrays();
/*compute partition function*/
if (pf) {
cofoldF AB, AA, BB;
FLT_OR_DBL *probs;
if (dangles==1) {
dangles=2; /* recompute with dangles as in pf_fold() */
min_en = energy_of_structure(rec_sequence, structure, 0);
dangles=1;
}
kT = (betaScale*((temperature+K0)*GASCONST))/1000.; /* in Kcal */
pf_scale = exp(-(sfact*min_en)/kT/length);
if (length>2000) fprintf(stderr, "scaling factor %f\n", pf_scale);
pf_parameters = get_boltzmann_factors(temperature, betaScale, md, pf_scale);
if (cstruc!=NULL)
strncpy(structure, cstruc, length+1);
AB = co_pf_fold_par(rec_sequence, structure, pf_parameters, do_backtrack, fold_constrained);
if (do_backtrack) {
char *costruc;
costruc = (char *) space(sizeof(char)*(strlen(structure)+2));
if (cut_point<0) printf("%s", structure);
else {
strncpy(costruc, structure, cut_point-1);
strcat(costruc, "&");
strcat(costruc, structure+cut_point-1);
printf("%s", costruc);
}
if (!istty) printf(" [%6.2f]\n", AB.FAB);
else printf("\n");/*8.6.04*/
}
if ((istty)||(!do_backtrack))
printf(" free energy of ensemble = %6.2f kcal/mol\n", AB.FAB);
printf(" frequency of mfe structure in ensemble %g",
exp((AB.FAB-min_en)/kT));
printf(" , delta G binding=%6.2f\n", AB.FcAB - AB.FA - AB.FB);
probs = export_co_bppm();
assign_plist_from_pr(&prAB, probs, length, bppmThreshold);
/* if (doQ) make_probsum(length,fname); */ /*compute prob of base paired*/
/* free_co_arrays(); */
if (doT) { /* cofold of all dimers, monomers */
int Blength, Alength;
char *Astring, *Bstring, *orig_Astring, *orig_Bstring;
char *Newstring;
char Newname[30];
char comment[80];
if (cut_point<0) {
printf("Sorry, i cannot do that with only one molecule, please give me two or leave it\n");
free(mfAB);
free(prAB);
continue;
}
if (dangles==1) dangles=2;
Alength=cut_point-1; /*length of first molecule*/
Blength=length-cut_point+1; /*length of 2nd molecule*/
Astring=(char *)space(sizeof(char)*(Alength+1));/*Sequence of first molecule*/
Bstring=(char *)space(sizeof(char)*(Blength+1));/*Sequence of second molecule*/
strncat(Astring,rec_sequence,Alength);
strncat(Bstring,rec_sequence+Alength,Blength);
orig_Astring=(char *)space(sizeof(char)*(Alength+1));/*Sequence of first molecule*/
orig_Bstring=(char *)space(sizeof(char)*(Blength+1));/*Sequence of second molecule*/
strncat(orig_Astring,orig_sequence,Alength);
strncat(orig_Bstring,orig_sequence+Alength,Blength);
/* compute AA dimer */
AA=do_partfunc(Astring, Alength, 2, &prAA, &mfAA, pf_parameters);
/* compute BB dimer */
BB=do_partfunc(Bstring, Blength, 2, &prBB, &mfBB, pf_parameters);
/*free_co_pf_arrays();*/
/* compute A monomer */
do_partfunc(Astring, Alength, 1, &prA, &mfA, pf_parameters);
/* compute B monomer */
do_partfunc(Bstring, Blength, 1, &prB, &mfB, pf_parameters);
compute_probabilities(AB.F0AB, AB.FA, AB.FB, prAB, prA, prB, Alength);
compute_probabilities(AA.F0AB, AA.FA, AA.FA, prAA, prA, prA, Alength);
compute_probabilities(BB.F0AB, BB.FA, BB.FA, prBB, prA, prB, Blength);
printf("Free Energies:\nAB\t\tAA\t\tBB\t\tA\t\tB\n%.6f\t%6f\t%6f\t%6f\t%6f\n",
AB.FcAB, AA.FcAB, BB.FcAB, AB.FA, AB.FB);
if (doC) {
do_concentrations(AB.FcAB, AA.FcAB, BB.FcAB, AB.FA, AB.FB, ConcAandB);
free(ConcAandB);/*freeen*/
}
if (fname[0]!='\0') {
strcpy(ffname, fname);
strcat(ffname, "_dp5.ps");
} else strcpy(ffname, "dot5.ps");
/*output of the 5 dot plots*/
/*AB dot_plot*/
/*write Free Energy into comment*/
sprintf(comment,"\n%%Heterodimer AB FreeEnergy= %.9f\n", AB.FcAB);
/*reset cut_point*/
cut_point=Alength+1;
/*write New name*/
strcpy(Newname,"AB");
strcat(Newname,ffname);
(void)PS_dot_plot_list(orig_sequence, Newname, prAB, mfAB, comment);
/*AA dot_plot*/
sprintf(comment,"\n%%Homodimer AA FreeEnergy= %.9f\n",AA.FcAB);
/*write New name*/
strcpy(Newname,"AA");
strcat(Newname,ffname);
/*write AA sequence*/
Newstring=(char*)space((2*Alength+1)*sizeof(char));
strcpy(Newstring,orig_Astring);
strcat(Newstring,orig_Astring);
(void)PS_dot_plot_list(Newstring, Newname, prAA, mfAA, comment);
free(Newstring);
/*BB dot_plot*/
sprintf(comment,"\n%%Homodimer BB FreeEnergy= %.9f\n",BB.FcAB);
/*write New name*/
strcpy(Newname,"BB");
strcat(Newname,ffname);
/*write BB sequence*/
Newstring=(char*)space((2*Blength+1)*sizeof(char));
strcpy(Newstring,orig_Bstring);
strcat(Newstring,orig_Bstring);
/*reset cut_point*/
cut_point=Blength+1;
(void)PS_dot_plot_list(Newstring, Newname, prBB, mfBB, comment);
free(Newstring);
/*A dot plot*/
/*reset cut_point*/
cut_point=-1;
sprintf(comment,"\n%%Monomer A FreeEnergy= %.9f\n",AB.FA);
/*write New name*/
strcpy(Newname,"A");
strcat(Newname,ffname);
/*write BB sequence*/
(void)PS_dot_plot_list(orig_Astring, Newname, prA, mfA, comment);
/*B monomer dot plot*/
sprintf(comment,"\n%%Monomer B FreeEnergy= %.9f\n",AB.FB);
/*write New name*/
strcpy(Newname,"B");
strcat(Newname,ffname);
/*write BB sequence*/
(void)PS_dot_plot_list(orig_Bstring, Newname, prB, mfB, comment);
free(Astring); free(Bstring); free(orig_Astring); free(orig_Bstring);
free(prAB); free(prAA); free(prBB); free(prA); free(prB);
free(mfAB); free(mfAA); free(mfBB); free(mfA); free(mfB);
} /*end if(doT)*/
free(pf_parameters);
}/*end if(pf)*/
if (do_backtrack) {
if (fname[0]!='\0') {
strcpy(ffname, fname);
strcat(ffname, "_dp.ps");
} else strcpy(ffname, "dot.ps");
if (!doT) {
if (pf) { (void) PS_dot_plot_list(rec_sequence, ffname, prAB, mfAB, "doof");
free(prAB);}
free(mfAB);
}
}
if (!doT) free_co_pf_arrays();
(void) fflush(stdout);
/* clean up */
if(cstruc) free(cstruc);
if(rec_id) free(rec_id);
free(rec_sequence);
free(orig_sequence);
free(structure);
/* free the rest of current dataset */
if(rec_rest){
for(i=0;rec_rest[i];i++) free(rec_rest[i]);
free(rec_rest);
}
rec_id = rec_sequence = orig_sequence = structure = cstruc = NULL;
rec_rest = NULL;
/* print user help for the next round if we get input from tty */
if(istty){
printf("Use '&' to connect 2 sequences that shall form a complex.\n");
if(fold_constrained){
print_tty_constraint(VRNA_CONSTRAINT_DOT | VRNA_CONSTRAINT_X | VRNA_CONSTRAINT_ANG_BRACK | VRNA_CONSTRAINT_RND_BRACK);
print_tty_input_seq_str("Input sequence (upper or lower case) followed by structure constraint\n");
}
else print_tty_input_seq();
}
}
return EXIT_SUCCESS;
}
int main(int argc, char *argv[]){
struct RNAeval_args_info args_info;
char *string, *structure, *orig_sequence, *tmp;
char *rec_sequence, *rec_id, **rec_rest;
char fname[FILENAME_MAX_LENGTH];
char *ParamFile;
int i, length1, length2;
float energy;
int istty;
int circular=0;
int noconv=0;
int verbose = 0;
unsigned int rec_type, read_opt;
string = orig_sequence = ParamFile = NULL;
gquad = 0;
dangles = 2;
/*
#############################################
# check the command line parameters
#############################################
*/
if(RNAeval_cmdline_parser (argc, argv, &args_info) != 0) exit(1);
/* temperature */
if(args_info.temp_given) temperature = args_info.temp_arg;
/* do not take special tetra loop energies into account */
if(args_info.noTetra_given) tetra_loop=0;
/* set dangle model */
if(args_info.dangles_given){
if((args_info.dangles_arg < 0) || (args_info.dangles_arg > 3))
warn_user("required dangle model not implemented, falling back to default dangles=2");
else
dangles = args_info.dangles_arg;
}
/* do not convert DNA nucleotide "T" to appropriate RNA "U" */
if(args_info.noconv_given) noconv = 1;
/* set energy model */
if(args_info.energyModel_given) energy_set = args_info.energyModel_arg;
/* take another energy parameter set */
if(args_info.paramFile_given) ParamFile = strdup(args_info.paramFile_arg);
/* assume RNA sequence to be circular */
if(args_info.circ_given) circular=1;
/* logarithmic multiloop energies */
if(args_info.logML_given) logML = 1;
/* be verbose */
if(args_info.verbose_given) verbose = 1;
/* gquadruplex support */
if(args_info.gquad_given) gquad = 1;
/* free allocated memory of command line data structure */
RNAeval_cmdline_parser_free (&args_info);
/*
#############################################
# begin initializing
#############################################
*/
if (ParamFile!=NULL) read_parameter_file(ParamFile);
rec_type = read_opt = 0;
rec_id = rec_sequence = NULL;
rec_rest = NULL;
istty = isatty(fileno(stdout)) && isatty(fileno(stdin));
if(circular && gquad){
nrerror("G-Quadruplex support is currently not available for circular RNA structures");
}
/* set options we wanna pass to read_record */
if(istty){
read_opt |= VRNA_INPUT_NOSKIP_BLANK_LINES;
print_tty_input_seq_str("Use '&' to connect 2 sequences that shall form a complex.\n"
"Input sequence (upper or lower case) followed by structure");
}
/*
#############################################
# main loop: continue until end of file
#############################################
*/
while(
!((rec_type = read_record(&rec_id, &rec_sequence, &rec_rest, read_opt))
& (VRNA_INPUT_ERROR | VRNA_INPUT_QUIT))){
if(rec_id){
if(!istty) printf("%s\n", rec_id);
(void) sscanf(rec_id, ">%" XSTR(FILENAME_ID_LENGTH) "s", fname);
}
else fname[0] = '\0';
cut_point = -1;
string = tokenize(rec_sequence);
length2 = (int) strlen(string);
tmp = extract_record_rest_structure((const char **)rec_rest, 0, (rec_id) ? VRNA_OPTION_MULTILINE : 0);
if(!tmp)
nrerror("structure missing");
structure = tokenize(tmp);
length1 = (int) strlen(structure);
if(length1 != length2)
nrerror("structure and sequence differ in length!");
free(tmp);
/* convert DNA alphabet to RNA if not explicitely switched off */
if(!noconv) str_DNA2RNA(string);
/* store case-unmodified sequence */
orig_sequence = strdup(string);
/* convert sequence to uppercase letters only */
str_uppercase(string);
if(istty){
if (cut_point == -1)
printf("length = %d\n", length1);
else
printf("length1 = %d\nlength2 = %d\n", cut_point-1, length1-cut_point+1);
}
if(gquad)
energy = energy_of_gquad_structure(string, structure, verbose);
else
energy = (circular) ? energy_of_circ_structure(string, structure, verbose) : energy_of_structure(string, structure, verbose);
if (cut_point == -1)
printf("%s\n%s", orig_sequence, structure);
else {
char *pstring, *pstruct;
pstring = costring(orig_sequence);
pstruct = costring(structure);
printf("%s\n%s", pstring, pstruct);
free(pstring);
free(pstruct);
}
if (istty)
printf("\n energy = %6.2f\n", energy);
else
printf(" (%6.2f)\n", energy);
/* clean up */
(void) fflush(stdout);
if(rec_id) free(rec_id);
free(rec_sequence);
free(structure);
/* free the rest of current dataset */
if(rec_rest){
for(i=0;rec_rest[i];i++) free(rec_rest[i]);
free(rec_rest);
}
rec_id = rec_sequence = structure = NULL;
rec_rest = NULL;
free(string);
free(orig_sequence);
string = orig_sequence = NULL;
/* print user help for the next round if we get input from tty */
if(istty){
print_tty_input_seq_str("Use '&' to connect 2 sequences that shall form a complex.\n"
"Input sequence (upper or lower case) followed by structure");
}
}
return EXIT_SUCCESS;
}
int main(int argc, char *argv[]) {
struct RNAfold_args_info args_info;
char *string, *input_string, *structure=NULL, *cstruc=NULL;
char fname[80], ffname[80], gfname[80], *ParamFile=NULL;
char *ns_bases=NULL, *c;
int i, j, ii, jj, mu, length, l, sym, r, pf=0, noconv=0;
unsigned int input_type;
double energy, min_en, kT, sfact=1.07;
int doMEA=0, circular = 0, N;
char *pf_struc;
double dist;
plist *pl;
FILE * filehandle;
FILE * statsfile;
char* line;
double tau = 0.01; /* Variance of energy parameters */
double sigma = 0.01; /* Variance of experimental constraints */
double *gradient; /* Gradient for steepest descent search
epsilon[i+1]= epsilon[i] - gradient *
step_size */
double initial_step_size = 0.5; /* Initial step size for steepest
descent search */
double step_size;
double D; /* Discrepancy (i.e. value of objective
function) for the current
prediction */
int iteration, max_iteration = 2000; /* Current and maximum number of
iterations after which
algorithm stops */
double precision = 0.1; /* cutoff used for stop conditions */
double tolerance = 0.1; /* Parameter used by various GSL minimizers */
int method_id = 1; /* Method to use for minimization, 0 and 1
are custom steepest descent, the rest
are GSL implementations (see below)*/
int initial_guess_method = 0;
int sample_N = 1000;
double *prev_epsilon;
double *prev_gradient;
double DD, prev_D, sum, norm;
int status;
double* gradient_numeric;
double* gradient_numeric_gsl;
/* Minimizer vars */
const gsl_multimin_fdfminimizer_type *T;
gsl_multimin_fdfminimizer *minimizer;
gsl_vector *minimizer_x;
gsl_vector *minimizer_g;
gsl_multimin_function_fdf minimizer_func;
minimizer_pars_struct minimizer_pars;
char *constraints;
char outfile[256];
char constraints_file[256];
char epsilon_file[256];
FILE* fh;
double last_non_nan_lnQ;
pf_overflow = 0;
pf_underflow = 0;
dangles=2;
do_backtrack = 1;
string = NULL;
noPS = 0;
outfile[0]='\0';
epsilon_file[0]='\0';
strcpy(psDir, "dotplots");
if(RNAfold_cmdline_parser (argc, argv, &args_info) != 0) exit(1);
/* RNAbpfold specific options */
if (args_info.tau_given) tau = args_info.tau_arg;
if (args_info.sigma_given) sigma = args_info.sigma_arg;
if (args_info.precision_given) precision = args_info.precision_arg;
if (args_info.step_given) initial_step_size = args_info.step_arg;
if (args_info.maxN_given) max_iteration = args_info.maxN_arg;
if (args_info.minimization_given) method_id = args_info.minimization_arg;
if (args_info.init_given) initial_guess_method = args_info.init_arg;
if (args_info.tolerance_given) tolerance = args_info.tolerance_arg;
if (args_info.outfile_given) strcpy(outfile, args_info.outfile_arg);
if (args_info.constraints_given) strcpy(constraints_file, args_info.constraints_arg);
if (args_info.epsilon_given) strcpy(epsilon_file, args_info.epsilon_arg);
if (args_info.sampleGradient_given) sample_conditionals=1;
if (args_info.hybridGradient_given) {
sample_conditionals=1;
hybrid_conditionals=1;
}
if (args_info.numericalGradient_given) numerical=1;
if (args_info.sampleStructure_given) sample_structure=1;
if (args_info.psDir_given) strcpy(psDir, args_info.psDir_arg);
if (args_info.sparsePS_given) sparsePS=args_info.sparsePS_arg;
if (args_info.gridSearch_given) grid_search = 1;
/* Generic RNAfold options */
if (args_info.temp_given) temperature = args_info.temp_arg;
if (args_info.reference_given) fold_constrained=1;
if (args_info.noTetra_given) tetra_loop=0;
if (args_info.dangles_given) dangles = args_info.dangles_arg;
if (args_info.noLP_given) noLonelyPairs = 1;
if (args_info.noGU_given) noGU = 1;
if (args_info.noClosingGU_given) no_closingGU = 1;
if (args_info.noconv_given) noconv = 1;
if (args_info.energyModel_given) energy_set = args_info.energyModel_arg;
if (args_info.paramFile_given) ParamFile = strdup(args_info.paramFile_arg);
if (args_info.nsp_given) ns_bases = strdup(args_info.nsp_arg);
if (args_info.pfScale_given) sfact = args_info.pfScale_arg;
if (args_info.noPS_given) noPS=1;
/* Create postscript directory */
if (!noPS) {
struct stat stat_p;
if (stat (psDir, &stat_p) != 0) {
if (mkdir(psDir, S_IRWXU|S_IROTH|S_IRGRP ) !=0) {
fprintf(stderr, "WARNING: Could not create directory: %s", psDir);
}
}
}
if (ParamFile != NULL) {
read_parameter_file(ParamFile);
}
if (ns_bases != NULL) {
nonstandards = space(33);
c=ns_bases;
i=sym=0;
if (*c=='-') {
sym=1;
c++;
}
while (*c!='\0') {
if (*c!=',') {
nonstandards[i++]=*c++;
nonstandards[i++]=*c;
if ((sym)&&(*c!=*(c-1))) {
nonstandards[i++]=*c;
nonstandards[i++]=*(c-1);
}
}
c++;
}
}
/*Read sequence*/
fname[0] = '\0';
while((input_type = get_input_line(&input_string, 0)) & VRNA_INPUT_FASTA_HEADER) {
(void) sscanf(input_string, "%42s", fname);
free(input_string);
}
length = (int) strlen(input_string);
string = strdup(input_string);
free(input_string);
structure = (char *) space((unsigned) length+1);
/* For testing purpose pass dot bracket structure of reference structure via -C */
if (fold_constrained) {
input_type = get_input_line(&input_string, VRNA_INPUT_NOSKIP_COMMENTS);
if(input_type & VRNA_INPUT_QUIT) {
exit(1);
}
else if((input_type & VRNA_INPUT_MISC) && (strlen(input_string) > 0)) {
cstruc = strdup(input_string);
free(input_string);
}
else warn_user("-C was given but reference structure is missing");
}
if(noconv) {
str_RNA2RNA(string);
} else {
str_DNA2RNA(string);
}
/* Allocating space */
epsilon = (double *) space(sizeof(double)*(length+1));
exp_pert = (double **)space(sizeof(double *)*(length+1));
perturbations = (double **)space(sizeof(double *)*(length+1));
prev_epsilon = (double *) space(sizeof(double)*(length+1));
gradient = (double *) space(sizeof(double)*(length+1));
gradient_numeric = (double *) space(sizeof(double)*(length+1));
gradient_numeric_gsl = (double *) space(sizeof(double)*(length+1));
prev_gradient = (double *) space(sizeof(double)*(length+1));
q_unpaired = (double *) space(sizeof(double)*(length+1));
p_unpaired_cond = (double **)space(sizeof(double *)*(length+1));
p_unpaired_cond_sampled = (double **)space(sizeof(double *)*(length+1));
p_pp = (double **)space(sizeof(double *)*(length+1));
p_unpaired = (double *) space(sizeof(double)*(length+1));
p_unpaired_tmp = (double *) space(sizeof(double)*(length+1));
for (i=0; i <= length; i++) {
epsilon[i] = gradient[i] = q_unpaired[i] = 0.0;
p_unpaired_cond[i] = (double *) space(sizeof(double)*(length+1));
p_unpaired_cond_sampled[i] = (double *) space(sizeof(double)*(length+1));
p_pp[i] = (double *) space(sizeof(double)*(length+1));
exp_pert[i] = (double *) space(sizeof(double)*(length+1));
perturbations[i] = (double *) space(sizeof(double)*(length+1));
for (j=0; j <= length; j++) {
p_pp[i][j]=p_unpaired_cond[i][j] = 0.0;
p_unpaired_cond_sampled[i][j] = 0.0;
}
}
/*** If file with perturbation vector epsilon is given we fold using
this epsilon and are done ***/
if (args_info.epsilon_given) {
plist *pl, *pl1,*pl2;
filehandle = fopen (epsilon_file,"r");
if (filehandle == NULL) {
nrerror("Could not open file with perturbation vector.");
}
i=1;
while (1) {
double t;
line = get_line(filehandle);
if (line == NULL) break;
if (i>length) nrerror("Too many values in perturbation vector file.");
if (sscanf(line, "%lf", &epsilon[i]) !=1) {
nrerror("Error while reading perturbation vector file.");
}
i++;
}
if (i-1 != length) {
nrerror("Too few values in perturbation vector file.");
}
init_pf_fold(length);
pf_fold_pb(string, NULL);
sprintf(fname,"%s/dot.ps", psDir);
pl1 = make_plist(length, 1e-5);
(void) PS_dot_plot_list_epsilon(string, fname, NULL, pl1, epsilon, "");
exit(0);
}
/*** Get constraints from reference structure or from external file ***/
/* Structure was given by -C */
if (fold_constrained) {
for (i=0; i<length; i++) {
if (cstruc[i] == '(' || cstruc[i] == ')') {
q_unpaired[i+1] = 0.0;
} else {
q_unpaired[i+1] = 1.0;
}
}
/*Read constraints from file*/
} else {
filehandle = fopen (constraints_file,"r");
if (filehandle == NULL) {
nrerror("No constraints given as dot bracket or wrong file name");
}
i=1;
while (1) {
double t;
line = get_line(filehandle);
if (line == NULL) break;
if (i>length) nrerror("Too many values in constraints.dat");
if (sscanf(line, "%lf", &q_unpaired[i]) !=1) {
nrerror("Error while reading constraints.dat");
}
i++;
}
if (i-1 != length) {
nrerror("Too few values in constraints.dat");
}
}
/* Create file handle */
if (outfile[0] !='\0') {
statsfile = fopen (outfile,"w");
} else {
statsfile = fopen ("stats.dat","w");
}
setvbuf(statsfile, NULL, _IONBF, 0);
if (!grid_search) {
fprintf(statsfile, "Iteration\tDiscrepancy\tNorm\tdfCount\tMEA\tSampled_structure\tSampled_energy\tSampled_distance\tEpsilon\ttimestamp\n");
} else {
/* If we do a grid search we have a different output. */
fprintf(statsfile, "Dummy\tm\tb\tdummy\tMEA\tepsilon\n");
}
if (statsfile == NULL) {
nrerror("Could not open stats.dat for writing.");
}
fprintf(stderr, "tau^2 = %.4f; sigma^2 = %.4f; precision = %.4f; tolerance = %.4f; step-size: %.4f\n\n",
tau, sigma, precision, tolerance, initial_step_size);
st_back=1;
min_en = fold(string, structure);
(void) fflush(stdout);
if (length>2000) free_arrays();
pf_struc = (char *) space((unsigned) length+1);
kT = (temperature+273.15)*1.98717/1000.; /* in Kcal */
pf_scale = exp(-(sfact*min_en)/kT/length);
/* Set up minimizer */
minimizer_x = gsl_vector_alloc (length+1);
minimizer_g = gsl_vector_alloc (length+1);
for (i=0; i <= length; i++) {
epsilon[i] = 0.0;
gsl_vector_set (minimizer_g, i, 0.0);
gsl_vector_set (minimizer_x, i, epsilon[i]);
}
minimizer_pars.length=length;
minimizer_pars.seq = string;
minimizer_pars.tau=tau;
minimizer_pars.sigma=sigma;
minimizer_pars.kT=kT;
minimizer_func.n = length+1;
minimizer_func.f = calculate_f;
minimizer_func.df = numerical ? calculate_df_numerically: calculate_df;
minimizer_func.fdf = calculate_fdf;
minimizer_func.params = &minimizer_pars;
//min_en = fold_pb(string, structure);
//fprintf(stderr, "%f", min_en);
//exit(0);
/* Calling test functions for debugging */
for (i=1; i <= length; i++) {
if (i%2==0) {
epsilon[i] = +0.2*i;
} else {
epsilon[i] = -0.2*i;
}
}
//test_folding(string, length);
/* //test_stochastic_backtracking(string, length); */
/* //test_gradient(minimizer_func, minimizer_pars); */
/* //test_gradient_sampling(minimizer_func, minimizer_pars); */
//exit(1);
count_df_evaluations=0;
/* Initial guess for epsilon */
if (initial_guess_method !=0 && initial_guess_method !=3) {
/* Vars for inital guess methods */
double m,b;
double* curr_epsilon;
double* best_epsilon;
double best_m, best_b, best_scale;
double curr_D;
double min_D = 999999999.0;
double inc = +0.25;
double cut;
if (initial_guess_method == 1) fprintf(stderr, "Mathew's constant perturbations\n");
if (initial_guess_method == 2) fprintf(stderr, "Perturbations proportional to q-p\n");
curr_epsilon = (double *) space(sizeof(double)*(length+1));
best_epsilon = (double *) space(sizeof(double)*(length+1));
last_non_nan_lnQ = min_en;
// Calculate p_unpaired for unperturbed state which we need later
// for the proportinal method
if (initial_guess_method == 2) {
init_pf_fold(length);
for (i=0; i <= length; i++) {
epsilon[i] = 0.0;
}
pf_fold_pb(string, NULL);
for (i = 1; i < length; i++) {
for (j = i+1; j<= length; j++) {
p_pp[i][j]=p_pp[j][i]=pr[iindx[i]-j];
}
}
get_pair_prob_vector(p_pp, p_unpaired_tmp, length, 1);
free_pf_arrays();
}
/* We do the same grid search as in the Mathews paper Fig. 4*/
for (m=0.25; m <=7.0; m+=0.25) {
// Weird way of writing this inner loop for the grid search. We
// traverse the grid without big jumps in the parameters to make
// sure that the updated scaling factor is accurate all the time.
inc*=-1;
for (b = inc < 0.0 ? 0.0 : -3.0; inc < 0.0 ? b >= -3.0 : b<= 0.0 ; b+=inc) {
// calculate cut point with x-axis and skip parameter pairs
// which give a cut point outside the range of
// q_unpaired (0 to 1). They gave frequently overflows and the
// idea is that we both want positive and negative perturbations
cut = exp( (-1) * b / m ) - 1;
fprintf(stderr, "\nm = %.2f, b = %.2f, cut=%.2f\n", m, b, cut);
if (cut > 1.0 || cut < 0.01) {
fprintf(stderr, "\nSkipping m = %.2f, b = %.2f\n", m, b);
continue;
}
/* Mathew's constant perturbations */
if (initial_guess_method == 1) {
for (i=0; i <= length; i++) {
/* We add epsilon to unpaired regions (as opposed to
paired regions as in the Mathews paper) so we multiply
by -1; if missing data we set it to 0.0 */
if (q_unpaired[i] < -0.5) {
curr_epsilon[i] = 0.0;
} else {
curr_epsilon[i] = (m *(log(q_unpaired[i]+1))+b) *(-1);
}
gsl_vector_set (minimizer_x, i, curr_epsilon[i]);
}
/* Perturbations proportional to q-p */
} else {
for (i=0; i <= length; i++) {
curr_epsilon[i] = (m *(log(q_unpaired[i]+1)-log(p_unpaired_tmp[i]+1))+ b ) * (-1);
gsl_vector_set (minimizer_x, i, curr_epsilon[i]);
}
}
// Repeat and adjust scaling factor until we get result without over-/underflows
do {
// First we use default scaling factor
if (pf_underflow == 0 && pf_overflow == 0) {
sfact = 1.070;
}
if (pf_underflow) {
sfact *= 0.8;
fprintf(stderr,"Underflow, adjusting sfact to %.4f\n", sfact );
}
if (pf_overflow) {
sfact *= 1.2;
fprintf(stderr,"Overflow, adjusting sfact to %.4f\n", sfact );
}
pf_scale = exp(-(sfact*last_non_nan_lnQ)/kT/length);
//fprintf(stderr,"Scaling factor is now: %.4e\n", pf_scale);
curr_D = calculate_f(minimizer_x, (void*)&minimizer_pars);
if (!isnan(last_lnQ)) last_non_nan_lnQ = last_lnQ;
// Give up when even extreme scaling does not give results
// (for some reason I could not get rid of overflows even with high scaling factors)
if (sfact < 0.1 || sfact > 2.0) break;
} while (pf_underflow == 1 || pf_overflow == 1);
// We have not given up so everything is ok now
if (!(sfact < 0.1 || sfact > 2.0)) {
if (curr_D < min_D) {
min_D = curr_D;
for (i=0; i <= length; i++) {
best_epsilon[i] = curr_epsilon[i];
}
best_m = m;
best_b = b;
best_scale = pf_scale;
}
/*If we are interested in the grid search we misuse the
print_stats function and report m and b together with MEA*/
if (grid_search) {
for (i=0; i <= length; i++) {
epsilon[i] = curr_epsilon[i];
}
print_stats(statsfile, string, cstruc, length, 0, 0, m, 0.0, b, 0);
}
fprintf(stderr, "curr D: %.2f, minimum D: %.2f\n", curr_D, min_D);
// Adjust pf_scale with default scaling factor but lnQ from
// previous step
sfact = 1.070;
pf_scale = exp(-(sfact*last_lnQ)/kT/length);
} else {
sfact = 1.070;
fprintf(stderr, "Skipping m = %.2f, b = %.2f; did not get stable result.\n", m, b);
}
} // for b
} // for m
fprintf(stderr, "Minimum found: m=%.2f, b=%.2f: %.2f\n", best_m, best_b, min_D);
for (i=0; i <= length; i++) {
epsilon[i] = best_epsilon[i];
gsl_vector_set (minimizer_x, i, best_epsilon[i]);
}
pf_scale = best_scale;
}
if (initial_guess_method == 3) {
srand((unsigned)time(0));
for (i=0; i <= length; i++) {
double r = (double)rand()/(double)RAND_MAX * 4 - 2;
epsilon[i] = r;
gsl_vector_set (minimizer_x, i, epsilon[i]);
}
}
/* If we just wanted a grid search we are done now. */
if (grid_search) {
exit(0);
}
prev_D = calculate_f(minimizer_x, (void*)&minimizer_pars);
print_stats(statsfile, string, cstruc, length, 0 , count_df_evaluations , prev_D, -1.0, 0.0,1);
/* GSL minimization */
if (method_id !=0) {
if (method_id > 2) {
char name[100];
// Available algorithms
// 3 gsl_multimin_fdfminimizer_conjugate_fr
// 4 gsl_multimin_fdfminimizer_conjugate_pr
// 5 gsl_multimin_fdfminimizer_vector_bfgs
// 6 gsl_multimin_fdfminimizer_vector_bfgs2
// 7 gsl_multimin_fdfminimizer_steepest_descent
// http://www.gnu.org/software/gsl/manual/html_node/Multimin-Algorithms-with-Derivatives.html
switch (method_id) {
case 2:
minimizer = gsl_multimin_fdfminimizer_alloc (gsl_multimin_fdfminimizer_conjugate_fr, length+1);
strcpy(name, "Fletcher-Reeves conjugate gradient");
break;
case 3:
minimizer = gsl_multimin_fdfminimizer_alloc (gsl_multimin_fdfminimizer_conjugate_pr, length+1);
strcpy(name, "Polak-Ribiere conjugate gradient");
break;
case 4:
minimizer = gsl_multimin_fdfminimizer_alloc ( gsl_multimin_fdfminimizer_vector_bfgs, length+1);
strcpy(name, "Broyden-Fletcher-Goldfarb-Shanno");
break;
case 5:
minimizer = gsl_multimin_fdfminimizer_alloc ( gsl_multimin_fdfminimizer_vector_bfgs2, length+1);
strcpy(name, "Broyden-Fletcher-Goldfarb-Shanno (improved version)");
break;
case 6:
minimizer = gsl_multimin_fdfminimizer_alloc (gsl_multimin_fdfminimizer_steepest_descent, length+1);
strcpy(name, "Gradient descent (GSL implmementation)");
break;
}
fprintf(stderr, "Starting minimization via GSL implementation of %s...\n\n", name);
// The last two parmeters are step size and tolerance (with
// different meaning for different algorithms
gsl_multimin_fdfminimizer_set (minimizer, &minimizer_func, minimizer_x, initial_step_size, tolerance);
iteration = 1;
do {
status = gsl_multimin_fdfminimizer_iterate (minimizer);
D = minimizer->f;
norm = gsl_blas_dnrm2(minimizer->gradient);
print_stats(statsfile, string, cstruc, length,iteration, count_df_evaluations, D, prev_D, norm, iteration%sparsePS == 0);
prev_D = D;
if (status) {
fprintf(stderr, "An unexpected error has occured in the iteration (status:%i)\n", status);
break;
}
status = gsl_multimin_test_gradient (minimizer->gradient, precision);
if (status == GSL_SUCCESS) fprintf(stderr, "Minimum found stopping.\n");
iteration++;
} while (status == GSL_CONTINUE && iteration < max_iteration);
gsl_multimin_fdfminimizer_free (minimizer);
gsl_vector_free (minimizer_x);
/* Custom implementation of steepest descent */
} else {
if (method_id == 1) {
fprintf(stderr, "Starting custom implemented steepest descent search...\n\n");
} else {
fprintf(stderr, "Starting custom implemented steepest descent search with Barzilai Borwein step size...\n\n");
}
iteration = 0;
D = 0.0;
while (iteration++ < max_iteration) {
for (i=1; i <= length; i++) {
gsl_vector_set (minimizer_x, i, epsilon[i]);
}
D = calculate_f(minimizer_x, (void*)&minimizer_pars);
if (numerical) {
calculate_df_numerically(minimizer_x, (void*)&minimizer_pars, minimizer_g);
} else {
calculate_df(minimizer_x, (void*)&minimizer_pars, minimizer_g);
}
for (i=1; i <= length; i++) {
gradient[i] = gsl_vector_get (minimizer_g, i);
}
// Do line search
fprintf(stderr, "\nLine search:\n");
// After the first iteration, use Barzilai-Borwain (1988) step size (currently turned off)
if (iteration>1 && method_id==2) {
double denominator=0.0;
double numerator=0.0;
for (i=1; i <= length; i++) {
numerator += (epsilon[i]-prev_epsilon[i]) * (gradient[i]-prev_gradient[i]);
denominator+=(gradient[i]-prev_gradient[i]) * (gradient[i]-prev_gradient[i]);
}
step_size = numerator / denominator;
norm =1.0;
} else {
// Use step sized given by the user (normalize it first)
step_size = initial_step_size / calculate_norm(gradient, length);
}
for (i=1; i <= length; i++) {
prev_epsilon[i] = epsilon[i];
prev_gradient[i] = gradient[i];
}
do {
for (mu=1; mu <= length; mu++) {
epsilon[mu] = prev_epsilon[mu] - step_size * gradient[mu];
}
for (i=1; i <= length; i++) {
gsl_vector_set (minimizer_x, i, epsilon[i]);
}
DD = calculate_f(minimizer_x, (void*)&minimizer_pars);
if (step_size > 0.0001) {
fprintf(stderr, "Old D: %.4f; New D: %.4f; Step size: %.4f\n", D, DD, step_size);
} else {
fprintf(stderr, "Old D: %.4f; New D: %.4f; Step size: %.4e\n", D, DD, step_size);
}
step_size /= 2;
} while (step_size > 1e-12 && DD > D);
norm = calculate_norm(gradient,length);
if (DD > D) {
fprintf(stderr, "Line search did not improve D in iteration %i. Stop.\n", iteration);
if (hybrid_conditionals) {
sample_conditionals=0;
} else {
break;
}
}
print_stats(statsfile, string, cstruc, length,iteration, count_df_evaluations, DD, prev_D, norm, iteration%sparsePS == 0);
if (norm<precision && iteration>1) {
fprintf(stderr, "Minimum found stopping.\n");
break;
}
prev_D = DD;
}
}
/* Force last dotplot to be printed */
print_stats(statsfile, string, cstruc, length,iteration, count_df_evaluations, DD, prev_D, norm, 1);
}
free(pf_struc);
if (cstruc!=NULL) free(cstruc);
(void) fflush(stdout);
free(string);
free(structure);
RNAfold_cmdline_parser_free (&args_info);
return 0;
}
/*--------------------------------------------------------------------------*/
int main(int argc, char *argv[]){
struct RNAplfold_args_info args_info;
unsigned int error = 0;
char fname[FILENAME_MAX_LENGTH], ffname[FILENAME_MAX_LENGTH], *c, *structure, *ParamFile, *ns_bases, *rec_sequence, *rec_id, **rec_rest, *orig_sequence;
unsigned int input_type;
int i, length, l, sym, r, istty, winsize, pairdist;
float cutoff;
int tempwin, temppair, tempunpaired;
FILE *pUfp = NULL, *spup = NULL;
double **pup = NULL; /*prob of being unpaired, lengthwise*/
int noconv, plexoutput, simply_putout, openenergies, binaries;
plist *pl, *dpp = NULL;
unsigned int rec_type, read_opt;
double betaScale;
pf_paramT *pf_parameters;
model_detailsT md;
dangles = 2;
cutoff = 0.01;
winsize = 70;
pairdist = 0;
unpaired = 0;
betaScale = 1.;
simply_putout = plexoutput = openenergies = noconv = 0;binaries=0;
tempwin = temppair = tempunpaired = 0;
structure = ParamFile = ns_bases = NULL;
rec_type = read_opt = 0;
rec_id = rec_sequence = orig_sequence = NULL;
rec_rest = NULL;
pf_parameters = NULL;
set_model_details(&md);
/*
#############################################
# check the command line parameters
#############################################
*/
if(RNAplfold_cmdline_parser (argc, argv, &args_info) != 0) exit(1);
/* temperature */
if(args_info.temp_given) temperature = args_info.temp_arg;
/* do not take special tetra loop energies into account */
if(args_info.noTetra_given) md.special_hp = tetra_loop=0;
/* set dangle model */
if(args_info.dangles_given){
if((args_info.dangles_arg != 0) && (args_info.dangles_arg != 2))
warn_user("required dangle model not implemented, falling back to default dangles=2");
else
md.dangles = dangles = args_info.dangles_arg;
}
/* do not allow weak pairs */
if(args_info.noLP_given) md.noLP = noLonelyPairs = 1;
/* do not allow wobble pairs (GU) */
if(args_info.noGU_given) md.noGU = noGU = 1;
/* do not allow weak closing pairs (AU,GU) */
if(args_info.noClosingGU_given) md.noGUclosure = no_closingGU = 1;
/* do not convert DNA nucleotide "T" to appropriate RNA "U" */
if(args_info.noconv_given) noconv = 1;
/* set energy model */
if(args_info.energyModel_given) energy_set = args_info.energyModel_arg;
/* take another energy parameter set */
if(args_info.paramFile_given) ParamFile = strdup(args_info.paramFile_arg);
/* Allow other pairs in addition to the usual AU,GC,and GU pairs */
if(args_info.nsp_given) ns_bases = strdup(args_info.nsp_arg);
/* set the maximum base pair span */
if(args_info.span_given) pairdist = args_info.span_arg;
/* set the pair probability cutoff */
if(args_info.cutoff_given) cutoff = args_info.cutoff_arg;
/* set the windowsize */
if(args_info.winsize_given) winsize = args_info.winsize_arg;
/* set the length of unstructured region */
if(args_info.ulength_given) unpaired = args_info.ulength_arg;
/* compute opening energies */
if(args_info.opening_energies_given) openenergies = 1;
/* print output on the fly */
if(args_info.print_onthefly_given) simply_putout = 1;
/* turn on RNAplex output */
if(args_info.plex_output_given) plexoutput = 1;
/* turn on binary output*/
if(args_info.binaries_given) binaries = 1;
if(args_info.betaScale_given) betaScale = args_info.betaScale_arg;
/* check for errorneous parameter options */
if((pairdist < 0) || (cutoff < 0.) || (unpaired < 0) || (winsize < 0)){
RNAplfold_cmdline_parser_print_help();
exit(EXIT_FAILURE);
}
/* free allocated memory of command line data structure */
RNAplfold_cmdline_parser_free(&args_info);
/*
#############################################
# begin initializing
#############################################
*/
if (ParamFile != NULL)
read_parameter_file(ParamFile);
if (ns_bases != NULL) {
nonstandards = space(33);
c=ns_bases;
i=sym=0;
if (*c=='-') {
sym=1; c++;
}
while (*c!='\0') {
if (*c!=',') {
nonstandards[i++]=*c++;
nonstandards[i++]=*c;
if ((sym)&&(*c!=*(c-1))) {
nonstandards[i++]=*c;
nonstandards[i++]=*(c-1);
}
}
c++;
}
}
/* check parameter options again and reset to reasonable values if needed */
if(openenergies && !unpaired) unpaired = 31;
if(pairdist == 0) pairdist = winsize;
if(pairdist > winsize){
fprintf(stderr, "pairdist (-L %d) should be <= winsize (-W %d);"
"Setting pairdist=winsize\n",pairdist, winsize);
pairdist = winsize;
}
if(dangles % 2){
warn_user("using default dangles = 2");
dangles = 2;
}
istty = isatty(fileno(stdout))&&isatty(fileno(stdin));
read_opt |= VRNA_INPUT_NO_REST;
if(istty){
print_tty_input_seq();
read_opt |= VRNA_INPUT_NOSKIP_BLANK_LINES;
}
/*
#############################################
# main loop: continue until end of file
#############################################
*/
while(
!((rec_type = read_record(&rec_id, &rec_sequence, &rec_rest, read_opt))
& (VRNA_INPUT_ERROR | VRNA_INPUT_QUIT))){
/*
########################################################
# init everything according to the data we've read
########################################################
*/
if(rec_id){
if(!istty) printf("%s\n", rec_id);
(void) sscanf(rec_id, ">%" XSTR(FILENAME_ID_LENGTH) "s", fname);
}
else fname[0] = '\0';
length = (int)strlen(rec_sequence);
structure = (char *) space((unsigned) length+1);
/* convert DNA alphabet to RNA if not explicitely switched off */
if(!noconv) str_DNA2RNA(rec_sequence);
/* store case-unmodified sequence */
orig_sequence = strdup(rec_sequence);
/* convert sequence to uppercase letters only */
str_uppercase(rec_sequence);
if(istty) printf("length = %d\n", length);
/*
########################################################
# done with 'stdin' handling
########################################################
*/
if(length > 1000000){
if(!simply_putout && !unpaired){
printf("Switched to simple output mode!!!\n");
simply_putout = 1;
}
}
if(unpaired && simply_putout){
printf("Output simplification not possible if unpaired is switched on\n");
simply_putout = 0;
}
/* restore winsize if altered before */
if(tempwin != 0){
winsize = tempwin;
tempwin = 0;
}
/* restore pairdist if altered before */
if(temppair != 0){
pairdist = temppair;
temppair = 0;
}
/* restore ulength if altered before */
if(tempunpaired != 0){
unpaired = tempunpaired;
tempunpaired = 0;
}
/* adjust winsize, pairdist and ulength if necessary */
if(length < winsize){
fprintf(stderr, "WARN: window size %d larger than sequence length %d\n", winsize, length);
tempwin = winsize;
winsize = length;
if (pairdist>winsize) {
temppair=pairdist;
pairdist=winsize;
}
if (unpaired>winsize) {
tempunpaired=unpaired;
unpaired=winsize;
}
}
/*
########################################################
# begin actual computations
########################################################
*/
if (length >= 5){
/* construct output file names */
char fname1[FILENAME_MAX_LENGTH], fname2[FILENAME_MAX_LENGTH], fname3[FILENAME_MAX_LENGTH], fname4[FILENAME_MAX_LENGTH], fname_t[FILENAME_MAX_LENGTH];
strcpy(fname_t, (fname[0] != '\0') ? fname : "plfold");
strcpy(fname1, fname_t);
strcpy(fname2, fname_t);
strcpy(fname3, fname_t);
strcpy(fname4, fname_t);
strcpy(ffname, fname_t);
strcat(fname1, "_lunp");
strcat(fname2, "_basepairs");
strcat(fname3, "_uplex");
if(binaries){
strcat(fname4, "_openen_bin");
}
else{
strcat(fname4, "_openen");
}
strcat(ffname, "_dp.ps");
pf_parameters = get_boltzmann_factors(temperature, betaScale, md, -1);
if(unpaired > 0){
pup =(double **) space((length+1)*sizeof(double *));
pup[0] =(double *) space(sizeof(double)); /*I only need entry 0*/
pup[0][0] = unpaired;
}
pUfp = spup = NULL;
if(simply_putout){
spup = fopen(fname2, "w");
pUfp = (unpaired > 0) ? fopen(fname1, "w") : NULL;
pl = pfl_fold_par(rec_sequence, winsize, pairdist, cutoff, pup, &dpp, pUfp, spup, pf_parameters);
if(pUfp != NULL) fclose(pUfp);
if(spup != NULL) fclose(spup);
}
else{
pl = pfl_fold_par(rec_sequence, winsize, pairdist, cutoff, pup, &dpp, pUfp, spup, pf_parameters);
PS_dot_plot_turn(orig_sequence, pl, ffname, pairdist);
if (unpaired > 0){
if(plexoutput){
pUfp = fopen(fname3, "w");
putoutphakim_u(pup,length, unpaired, pUfp);
fclose(pUfp);
}
pUfp = fopen(openenergies ? fname4 : fname1, "w");
if(binaries){
putoutpU_prob_bin_par(pup, length, unpaired, pUfp, openenergies, pf_parameters);
}
else{
putoutpU_prob_par(pup, length, unpaired, pUfp, openenergies, pf_parameters);
}
fclose(pUfp);
}
}
if(pl) free(pl);
if(unpaired > 0){
free(pup[0]);
free(pup);
}
free(pf_parameters);
}
(void) fflush(stdout);
/* clean up */
if(rec_id) free(rec_id);
free(rec_sequence);
free(orig_sequence);
free(structure);
rec_id = rec_sequence = orig_sequence = NULL;
rec_rest = NULL;
/* print user help for the next round if we get input from tty */
if(istty) print_tty_input_seq();
}
return EXIT_SUCCESS;
}
int main(int argc, char *argv[]){
struct RNAsubopt_args_info args_info;
unsigned int input_type;
char fname[80], *cstruc, *sequence, *c, *input_string;
char *structure = NULL, *ParamFile = NULL, *ns_bases = NULL;
int i, length, l, sym, istty;
double deltaf, deltap;
int delta, n_back, noconv, circular, dos, zuker;
do_backtrack = 1;
dangles = 2;
delta = 100;
deltap = n_back = noconv = circular = dos = zuker = 0;
/*
#############################################
# check the command line parameters
#############################################
*/
if(RNAsubopt_cmdline_parser (argc, argv, &args_info) != 0) exit(1);
/* temperature */
if(args_info.temp_given) temperature = args_info.temp_arg;
/* structure constraint */
if(args_info.constraint_given) fold_constrained=1;
/* do not take special tetra loop energies into account */
if(args_info.noTetra_given) tetra_loop=0;
/* set dangle model */
if(args_info.dangles_given) dangles = args_info.dangles_arg;
/* do not allow weak pairs */
if(args_info.noLP_given) noLonelyPairs = 1;
/* do not allow wobble pairs (GU) */
if(args_info.noGU_given) noGU = 1;
/* do not allow weak closing pairs (AU,GU) */
if(args_info.noClosingGU_given) no_closingGU = 1;
/* do not convert DNA nucleotide "T" to appropriate RNA "U" */
if(args_info.noconv_given) noconv = 1;
/* take another energy parameter set */
if(args_info.paramFile_given) ParamFile = strdup(args_info.paramFile_arg);
/* Allow other pairs in addition to the usual AU,GC,and GU pairs */
if(args_info.nsp_given) ns_bases = strdup(args_info.nsp_arg);
/* energy range */
if(args_info.deltaEnergy_given) delta = (int) (0.1+args_info.deltaEnergy_arg*100);
/* energy range after post evaluation */
if(args_info.deltaEnergyPost_given) deltap = args_info.deltaEnergyPost_arg;
/* sorted output */
if(args_info.sorted_given) subopt_sorted = 1;
/* assume RNA sequence to be circular */
if(args_info.circ_given) circular=1;
/* stochastic backtracking */
if(args_info.stochBT_given){
n_back = args_info.stochBT_arg;
init_rand();
}
/* density of states */
if(args_info.dos_given){
dos = 1;
print_energy = -999999;
}
/* logarithmic multiloop energies */
if(args_info.logML_given) logML = 1;
/* zuker subopts */
if(args_info.zuker_given) zuker = 1;
if(zuker){
if(circular){
warn_user("Sorry, zuker subopts not yet implemented for circfold");
RNAsubopt_cmdline_parser_print_help();
exit(1);
}
else if(n_back>0){
warn_user("Can't do zuker subopts and stochastic subopts at the same time");
RNAsubopt_cmdline_parser_print_help();
exit(1);
}
}
/* free allocated memory of command line data structure */
RNAsubopt_cmdline_parser_free(&args_info);
/*
#############################################
# begin initializing
#############################################
*/
if (ParamFile != NULL) read_parameter_file(ParamFile);
if (ns_bases != NULL) {
nonstandards = space(33);
c=ns_bases;
i=sym=0;
if (*c=='-') {
sym=1; c++;
}
while (*c!='\0') {
if (*c!=',') {
nonstandards[i++]=*c++;
nonstandards[i++]=*c;
if ((sym)&&(*c!=*(c-1))) {
nonstandards[i++]=*c;
nonstandards[i++]=*(c-1);
}
}
c++;
}
}
istty = isatty(fileno(stdout))&&isatty(fileno(stdin));
if(fold_constrained && istty) print_tty_constraint(VRNA_CONSTRAINT_DOT | VRNA_CONSTRAINT_X);
/*
#############################################
# main loop: continue until end of file
#############################################
*/
do {
cut_point = -1;
/*
########################################################
# handle user input from 'stdin'
########################################################
*/
if(istty){
if (!zuker)
printf("Use '&' to connect 2 sequences that shall form a complex.\n");
print_tty_input_seq();
}
/* extract filename from fasta header if available */
fname[0] = '\0';
while((input_type = get_input_line(&input_string, 0)) == VRNA_INPUT_FASTA_HEADER){
printf(">%s\n", input_string);
(void) sscanf(input_string, "%42s", fname);
free(input_string);
}
/* break on any error, EOF or quit request */
if(input_type & (VRNA_INPUT_QUIT | VRNA_INPUT_ERROR)){ break;}
/* else assume a proper sequence of letters of a certain alphabet (RNA, DNA, etc.) */
else{
sequence = tokenize(input_string); /* frees input_string */
length = (int) strlen(sequence);
}
structure = (char *) space((unsigned) length+1);
if(noconv) str_RNA2RNA(sequence);
else str_DNA2RNA(sequence);
if(istty){
if (cut_point == -1)
printf("length = %d\n", length);
else
printf("length1 = %d\nlength2 = %d\n", cut_point-1, length-cut_point+1);
}
/* get structure constraint or break if necessary, entering an empty line results in a warning */
if (fold_constrained) {
input_type = get_input_line(&input_string, VRNA_INPUT_NOSKIP_COMMENTS);
if(input_type & VRNA_INPUT_QUIT){ break;}
else if((input_type & VRNA_INPUT_MISC) && (strlen(input_string) > 0)){
cstruc = tokenize(input_string);
strncpy(structure, cstruc, length);
for (i=0; i<length; i++)
if (structure[i]=='|')
nrerror("constraints of type '|' not allowed");
free(cstruc);
}
else warn_user("constraints missing");
}
/*
########################################################
# done with 'stdin' handling, now init everything properly
########################################################
*/
if((logML != 0 || dangles==1 || dangles==3) && dos == 0)
if(deltap<=0) deltap = delta/100. + 0.001;
if (deltap>0)
print_energy = deltap;
/* first lines of output (suitable for sort +1n) */
if (fname[0] != '\0')
printf("> %s [%d]\n", fname, delta);
/* stochastic backtracking */
if(n_back>0){
double mfe, kT;
char *ss;
st_back=1;
ss = (char *) space(strlen(sequence)+1);
strncpy(ss, structure, length);
mfe = fold(sequence, ss);
kT = (temperature+273.15)*1.98717/1000.; /* in Kcal */
pf_scale = exp(-(1.03*mfe)/kT/length);
strncpy(ss, structure, length);
/* ignore return value, we are not interested in the free energy */
(circular) ? (void) pf_circ_fold(sequence, ss) : (void) pf_fold(sequence, ss);
free(ss);
for (i=0; i<n_back; i++) {
char *s;
s =(circular) ? pbacktrack_circ(sequence) : pbacktrack(sequence);
printf("%s\n", s);
free(s);
}
free_pf_arrays();
}
/* normal subopt */
else if(!zuker){
(circular) ? subopt_circ(sequence, structure, delta, stdout) : subopt(sequence, structure, delta, stdout);
if (dos) {
int i;
for (i=0; i<= MAXDOS && i<=delta/10; i++) {
printf("%4d %6d\n", i, density_of_states[i]);
}
}
}
/* Zuker suboptimals */
else{
SOLUTION *zr;
int i;
if (cut_point!=-1) {
nrerror("Sorry, zuker subopts not yet implemented for cofold\n");
}
zr = zukersubopt(sequence);
putoutzuker(zr);
(void)fflush(stdout);
for (i=0; zr[i].structure; i++) {
free(zr[i].structure);
}
free(zr);
}
(void)fflush(stdout);
free(sequence);
free(structure);
} while (1);
return 0;
}
int main(int argc, char *argv[]){
struct RNAheat_args_info args_info;
char *string, *input_string, *ns_bases, *c, *ParamFile, *rec_sequence, *rec_id, **rec_rest, *orig_sequence;
int i, length, l, sym;
float T_min, T_max, h;
int mpoints, istty, noconv = 0;
unsigned int input_type;
unsigned int rec_type, read_opt;
string = ParamFile = ns_bases = NULL;
T_min = 0.;
T_max = 100.;
h = 1;
mpoints = 2;
dangles = 2; /* dangles can be 0 (no dangles) or 2, default is 2 */
rec_type = read_opt = 0;
rec_id = rec_sequence = orig_sequence = NULL;
rec_rest = NULL;
/*
#############################################
# check the command line parameters
#############################################
*/
if(RNAheat_cmdline_parser(argc, argv, &args_info) != 0) exit(1);
/* do not take special tetra loop energies into account */
if(args_info.noTetra_given) tetra_loop=0;
/* set dangle model */
if(args_info.dangles_given){
if((args_info.dangles_arg != 0) && (args_info.dangles_arg != 2))
warn_user("required dangle model not implemented, falling back to default dangles=2");
else
dangles = args_info.dangles_arg;
}
/* do not allow weak pairs */
if(args_info.noLP_given) noLonelyPairs = 1;
/* do not allow wobble pairs (GU) */
if(args_info.noGU_given) noGU = 1;
/* do not allow weak closing pairs (AU,GU) */
if(args_info.noClosingGU_given) no_closingGU = 1;
/* do not convert DNA nucleotide "T" to appropriate RNA "U" */
if(args_info.noconv_given) noconv = 1;
/* set energy model */
if(args_info.energyModel_given) energy_set = args_info.energyModel_arg;
/* take another energy parameter set */
if(args_info.paramFile_given) ParamFile = strdup(args_info.paramFile_arg);
/* Allow other pairs in addition to the usual AU,GC,and GU pairs */
if(args_info.nsp_given) ns_bases = strdup(args_info.nsp_arg);
/* Tmin */
if(args_info.Tmin_given) T_min = args_info.Tmin_arg;
/* Tmax */
if(args_info.Tmax_given) T_max = args_info.Tmax_arg;
/* step size */
if(args_info.stepsize_given) h = args_info.stepsize_arg;
/* ipoints */
if(args_info.ipoints_given){
mpoints = args_info.ipoints_arg;
if (mpoints < 1) mpoints = 1;
if (mpoints > 100) mpoints = 100;
}
/* free allocated memory of command line data structure */
RNAheat_cmdline_parser_free (&args_info);
/*
#############################################
# begin initializing
#############################################
*/
if (ParamFile!=NULL) read_parameter_file(ParamFile);
if (ns_bases != NULL) {
nonstandards = space(33);
c=ns_bases;
i=sym=0;
if (*c=='-') {
sym=1; c++;
}
while (*c!='\0') {
if (*c!=',') {
nonstandards[i++]=*c++;
nonstandards[i++]=*c;
if ((sym)&&(*c!=*(c-1))) {
nonstandards[i++]=*c;
nonstandards[i++]=*(c-1);
}
}
c++;
}
}
istty = isatty(fileno(stdout))&&isatty(fileno(stdin));
read_opt |= VRNA_INPUT_NO_REST;
if(istty){
print_tty_input_seq();
read_opt |= VRNA_INPUT_NOSKIP_BLANK_LINES;
}
/*
#############################################
# main loop: continue until end of file
#############################################
*/
while(
!((rec_type = read_record(&rec_id, &rec_sequence, &rec_rest, read_opt))
& (VRNA_INPUT_ERROR | VRNA_INPUT_QUIT))){
/*
########################################################
# init everything according to the data we've read
########################################################
*/
if(rec_id && !istty) printf("%s\n", rec_id);
length = (int)strlen(rec_sequence);
/* convert DNA alphabet to RNA if not explicitely switched off */
if(!noconv) str_DNA2RNA(rec_sequence);
/* store case-unmodified sequence */
orig_sequence = strdup(rec_sequence);
/* convert sequence to uppercase letters only */
str_uppercase(rec_sequence);
if(istty) printf("length = %d\n", length);
/*
########################################################
# done with 'stdin' handling
########################################################
*/
heat_capacity(rec_sequence, T_min, T_max, h, mpoints);
(void) fflush(stdout);
/* clean up */
if(rec_id) free(rec_id);
free(rec_sequence);
free(orig_sequence);
rec_id = rec_sequence = orig_sequence = NULL;
rec_rest = NULL;
/* print user help for the next round if we get input from tty */
if(istty) print_tty_input_seq();
}
return EXIT_SUCCESS;
}
int main(int argc, char *argv[]){
struct RNAheat_args_info args_info;
char *string, *input_string, *ns_bases, *c, *ParamFile;
int i, length, l, sym;
float T_min, T_max, h;
int mpoints, istty, noconv = 0;
unsigned int input_type;
string = ParamFile = ns_bases = NULL;
T_min = 0.;
T_max = 100.;
h = 1;
mpoints = 2;
dangles = 2; /* dangles can be 0 (no dangles) or 2, default is 2 */
/*
#############################################
# check the command line parameters
#############################################
*/
if(RNAheat_cmdline_parser(argc, argv, &args_info) != 0) exit(1);
/* do not take special tetra loop energies into account */
if(args_info.noTetra_given) tetra_loop=0;
/* set dangle model */
if(args_info.dangles_given){
dangles = args_info.dangles_arg;
if(dangles % 2){
warn_user("using default dangles = 2");
dangles = 2;
}
}
/* do not allow weak pairs */
if(args_info.noLP_given) noLonelyPairs = 1;
/* do not allow wobble pairs (GU) */
if(args_info.noGU_given) noGU = 1;
/* do not allow weak closing pairs (AU,GU) */
if(args_info.noClosingGU_given) no_closingGU = 1;
/* do not convert DNA nucleotide "T" to appropriate RNA "U" */
if(args_info.noconv_given) noconv = 1;
/* set energy model */
if(args_info.energyModel_given) energy_set = args_info.energyModel_arg;
/* take another energy parameter set */
if(args_info.paramFile_given) ParamFile = strdup(args_info.paramFile_arg);
/* Allow other pairs in addition to the usual AU,GC,and GU pairs */
if(args_info.nsp_given) ns_bases = strdup(args_info.nsp_arg);
/* Tmin */
if(args_info.Tmin_given) T_min = args_info.Tmin_arg;
/* Tmax */
if(args_info.Tmax_given) T_max = args_info.Tmax_arg;
/* step size */
if(args_info.stepsize_given) h = args_info.stepsize_arg;
/* ipoints */
if(args_info.ipoints_given){
mpoints = args_info.ipoints_arg;
if (mpoints < 1) mpoints = 1;
if (mpoints > 100) mpoints = 100;
}
/* free allocated memory of command line data structure */
RNAheat_cmdline_parser_free (&args_info);
/*
#############################################
# begin initializing
#############################################
*/
if (ParamFile!=NULL) read_parameter_file(ParamFile);
if (ns_bases != NULL) {
nonstandards = space(33);
c=ns_bases;
i=sym=0;
if (*c=='-') {
sym=1; c++;
}
while (*c!='\0') {
if (*c!=',') {
nonstandards[i++]=*c++;
nonstandards[i++]=*c;
if ((sym)&&(*c!=*(c-1))) {
nonstandards[i++]=*c;
nonstandards[i++]=*(c-1);
}
}
c++;
}
}
istty = isatty(fileno(stdout))&&isatty(fileno(stdin));
/*
#############################################
# main loop: continue until end of file
#############################################
*/
do {
/*
########################################################
# handle user input from 'stdin'
########################################################
*/
if(istty) print_tty_input_seq();
/* skip fasta headers and comments */
while((input_type = get_input_line(&input_string, 0)) & VRNA_INPUT_FASTA_HEADER){
printf(">%s\n", input_string);
/* skip fasta headers and comments */
free(input_string);
}
/* break on any error, EOF or quit request */
if(input_type & (VRNA_INPUT_QUIT | VRNA_INPUT_ERROR)){ break;}
/* else assume a proper sequence of letters of a certain alphabet (RNA, DNA, etc.) */
else{
string = strdup(input_string);
length = (int) strlen(string);
free(input_string);
}
if(noconv) str_RNA2RNA(string);
else str_DNA2RNA(string);
if (istty) printf("length = %d\n", length);
heat_capacity(string, T_min, T_max, h, mpoints);
free(string);
(void) fflush(stdout);
} while (1);
return 0;
}
int main(int argc, char *argv[]){
struct RNALfold_args_info args_info;
char *input_string, *c, *string, *structure, *ParamFile, *ns_bases;
int i, length, l, sym, r, istty, noconv, maxdist;
double energy, min_en;
unsigned int input_type;
string = structure = ParamFile = ns_bases = NULL;
do_backtrack = 1;
noconv = 0;
maxdist = 150;
/*
#############################################
# check the command line parameters
#############################################
*/
if(RNALfold_cmdline_parser (argc, argv, &args_info) != 0) exit(1);
/* temperature */
if(args_info.temp_given) temperature = args_info.temp_arg;
/* do not take special tetra loop energies into account */
if(args_info.noTetra_given) tetra_loop=0;
/* set dangle model */
if(args_info.dangles_given) dangles = args_info.dangles_arg;
/* do not allow weak pairs */
if(args_info.noLP_given) noLonelyPairs = 1;
/* do not allow wobble pairs (GU) */
if(args_info.noGU_given) noGU = 1;
/* do not allow weak closing pairs (AU,GU) */
if(args_info.noClosingGU_given) no_closingGU = 1;
/* do not convert DNA nucleotide "T" to appropriate RNA "U" */
if(args_info.noconv_given) noconv = 1;
/* set energy model */
if(args_info.energyModel_given) energy_set = args_info.energyModel_arg;
/* take another energy parameter set */
if(args_info.paramFile_given) ParamFile = strdup(args_info.paramFile_arg);
/* Allow other pairs in addition to the usual AU,GC,and GU pairs */
if(args_info.nsp_given) ns_bases = strdup(args_info.nsp_arg);
/* set the maximum base pair span */
if(args_info.span_given) maxdist = args_info.span_arg;
/* check for errorneous parameter options */
if(maxdist < 0){
RNALfold_cmdline_parser_print_help();
exit(EXIT_FAILURE);
}
/* free allocated memory of command line data structure */
RNALfold_cmdline_parser_free (&args_info);
/*
#############################################
# begin initializing
#############################################
*/
if (ParamFile != NULL)
read_parameter_file(ParamFile);
if (ns_bases != NULL) {
nonstandards = space(33);
c=ns_bases;
i=sym=0;
if (*c=='-') {
sym=1; c++;
}
while (*c!='\0') {
if (*c!=',') {
nonstandards[i++]=*c++;
nonstandards[i++]=*c;
if ((sym)&&(*c!=*(c-1))) {
nonstandards[i++]=*c;
nonstandards[i++]=*(c-1);
}
}
c++;
}
}
istty = isatty(fileno(stdout))&&isatty(fileno(stdin));
/*
#############################################
# main loop: continue until end of file
#############################################
*/
do{
/*
########################################################
# handle user input from 'stdin'
########################################################
*/
if(istty) print_tty_input_seq();
/* skip fasta header and comment lines */
while((input_type = get_input_line(&input_string, 0)) & VRNA_INPUT_FASTA_HEADER){
printf(">%s\n", input_string);
free(input_string);
}
/* break on any error, EOF or quit request */
if(input_type & (VRNA_INPUT_QUIT | VRNA_INPUT_ERROR)){ break;}
/* else assume a proper sequence of letters of a certain alphabet (RNA, DNA, etc.) */
else{
length = (int) strlen(input_string);
string = strdup(input_string);
free(input_string);
}
printf("%s\n", string);
structure = (char *) space((unsigned) length+1);
if(noconv) str_RNA2RNA(string);
else str_DNA2RNA(string);
if(istty) printf("length = %d\n", length);
/*
########################################################
# done with 'stdin' handling
########################################################
*/
/* initialize_fold(length); */
update_fold_params();
min_en = Lfold((const char *)string, structure, maxdist);
printf("%s", structure);
if (istty)
printf("\n minimum free energy = %6.2f kcal/mol\n", min_en);
else
printf(" (%6.2f)\n", min_en);
(void) fflush(stdout);
free(string);
free(structure);
} while (1);
return 0;
}