Provided by: vienna-rna_2.7.0+dfsg-1_amd64 
NAME
RNAmultifold - manual page for RNAmultifold 2.7.0
SYNOPSIS
RNAmultifold [OPTION]... [FILE]...
DESCRIPTION
RNAmultifold 2.7.0
Compute secondary structures of multiple interacting RNAs
The program works much like RNAfold, but allows one to specify multiple RNA sequences which are then
allowed to form conncected components. RNA sequences are read from stdin in the usual format, i.e. each
line of input corresponds to one sequence, except for lines starting with ">" which contain the name of
the next sequence(s). Multiple strands must be concatenated using the \'&\' character as separator.
RNAmultifold can compute MFE, partition function, corresponding ensemble free energy and base pairing
probabilities. These properties are either computed for a particular arrangement (concatenation) of
sequences, for the full ensemble of the complex of input RNAs, or all complexes formed by the input
sequences up to a specified number of interacting sequences. Output consists of a PostScript "dot plot"
file containing the pair probabilities, see the RNAfold man page for details. The program will continue
to read new sequences until a line consisting of the single character '@' or an end of file condition is
encountered.
-h, --help
Print help and exit
--detailed-help
Print help, including all details and hidden options, and exit
--full-help
Print help, including hidden options, and exit
-V, --version
Print version and exit
-v, --verbose
Be verbose. (default=off)
Lower the log level setting such that even INFO messages are passed through.
I/O Options:
Command line options for input and output (pre-)processing
-j, --jobs[=number]
Split batch input into jobs and start processing in parallel using multiple threads. A value of 0
indicates to use as many parallel threads as computation cores are available.
(default=`0')
Default processing of input data is performed in a serial fashion, i.e. one sequence pair at a
time. Using this switch, a user can instead start the computation for many sequence pairs in the
input in parallel. RNAmultifold will create as many parallel computation slots as specified and
assigns input sequences of the input file(s) to the available slots. Note, that this increases
memory consumption since input alignments have to be kept in memory until an empty compute slot is
available and each running job requires its own dynamic programming matrices.
--unordered
Do not try to keep output in order with input while parallel processing is in place.
(default=off)
When parallel input processing (--jobs flag) is enabled, the order in which input is processed
depends on the host machines job scheduler. Therefore, any output to stdout or files generated by
this program will most likely not follow the order of the corresponding input data set. The
default of RNAmultifold is to use a specialized data structure to still keep the results output in
order with the input data. However, this comes with a trade-off in terms of memory consumption,
since all output must be kept in memory for as long as no chunks of consecutive, ordered output
are available. By setting this flag, RNAmultifold will not buffer individual results but print
them as soon as they have been computated.
--noconv
Do not automatically substitute nucleotide "T" with "U".
(default=off)
--auto-id
Automatically generate an ID for each sequence. (default=off)
The default mode of RNAmultifold is to automatically determine an ID from the input sequence data
if the input file format allows to do that. Sequence IDs are usually given in the FASTA header of
input sequences. If this flag is active, RNAmultifold ignores any IDs retrieved from the input and
automatically generates an ID for each sequence. This ID consists of a prefix and an increasing
number. This flag can also be used to add a FASTA header to the output even if the input has none.
--id-prefix=STRING
Prefix for automatically generated IDs (as used in output file names).
(default=`sequence')
If this parameter is set, each sequence will be prefixed with the provided string. Hence, the
output files will obey the following naming scheme: "prefix_xxxx_ss.ps" (secondary structure
plot), "prefix_xxxx_dp.ps" (dot-plot), "prefix_xxxx_dp2.ps" (stack probabilities), etc. where xxxx
is the sequence number. Note: Setting this parameter implies --auto-id.
--id-delim=CHAR
Change the delimiter between prefix and increasing number for automatically generated IDs (as used
in output file names).
(default=`_')
This parameter can be used to change the default delimiter "_" between the prefix string and the
increasing number for automatically generated ID.
--id-digits=INT
Specify the number of digits of the counter in automatically generated alignment IDs.
(default=`4')
When alignments IDs are automatically generated, they receive an increasing number, starting with
1. This number will always be left-padded by leading zeros, such that the number takes up a
certain width. Using this parameter, the width can be specified to the users need. We allow
numbers in the range [1:18]. This option implies --auto-id.
--id-start=LONG
Specify the first number in automatically generated IDs.
(default=`1')
When sequence IDs are automatically generated, they receive an increasing number, usually starting
with 1. Using this parameter, the first number can be specified to the users requirements. Note:
negative numbers are not allowed. Note: Setting this parameter implies to ignore any IDs
retrieved from the input data, i.e. it activates the --auto-id flag.
--filename-delim=CHAR
Change the delimiting character used in sanitized filenames.
(default=`ID-delimiter')
This parameter can be used to change the delimiting character used while sanitizing filenames,
i.e. replacing invalid characters. Note, that the default delimiter ALWAYS is the first character
of the "ID delimiter" as supplied through the --id-delim option. If the delimiter is a whitespace
character or empty, invalid characters will be simply removed rather than substituted. Currently,
we regard the following characters as illegal for use in filenames: backslash '\', slash '/',
question mark '?', percent sign '%', asterisk '*', colon ':', pipe symbol '|', double quote '"',
triangular brackets '<' and '>'.
--filename-full
Use full FASTA header to create filenames. (default=off)
This parameter can be used to deactivate the default behavior of limiting output filenames to the
first word of the sequence ID. Consider the following example: An input with FASTA header
'>NM_0001 Homo Sapiens some gene' usually produces output files with the prefix "NM_0001" without
the additional data available in the FASTA header, e.g. "NM_0001_ss.ps" for secondary structure
plots. With this flag set, no truncation of the output filenames is done, i.e. output filenames
receive the full FASTA header data as prefixes. Note, however, that invalid characters (such as
whitespace) will be substituted by a delimiting character or simply removed, (see also the
parameter option --filename-delim).
--log-level=level
Set log level threshold. (default=`2')
By default, any log messages are filtered such that only warnings (level 2) or errors (level 3)
are printed. This setting allows for specifying the log level threshold, where higher values
result in fewer information. Log-level 5 turns off all messages, even errors and other critical
information.
--log-file[=filename]
Print log messages to a file instead of stderr. (default=`RNAmultifold.log')
--log-time
Include time stamp in log messages.
(default=off)
--log-call
Include file and line of log calling function.
(default=off)
Algorithms:
Select additional algorithms which should be included in the calculations. The Minimum free
energy (MFE) and a structure representative are calculated in any case.
-p, --partfunc[=INT]
Calculate the partition function and base pairing probability matrix in addition to the MFE
structure. Default is calculation of mfe structure only.
(default=`1')
In addition to the MFE structure we print a coarse representation of the pair probabilities in
form of a pseudo bracket notation, followed by the ensemble free energy. Note that unless you
also specify -d2 or -d0, the partition function and mfe calculations will use a slightly different
energy model. See the discussion of dangling end options below.
An additionally passed value to this option changes the behavior of partition function
calculation:
In order to calculate the partition function but not the pair probabilities
use the -p0 option and save about
50% in runtime. This prints the ensemble free energy 'dG=-kT ln(Z)'.
-a, --all_pf[=INT]
Compute the partition function and free energies not only for the complex formed by the input
sequences (the "ABC... mutimer"), but also of all complexes formed by the input sequences up to
the number of input sequences, e.g. AAA, AAB, ABB, BBB, etc.
(default=`1')
The output will contain the free energies for each of these species. Using -a automatically
switches on the -p option.
-c, --concentrations
In addition to everything listed under the -a option, read in initial monomer concentrations and
compute the expected equilibrium concentrations of all possible species (A, B, AA, BB, AB, etc).
(default=off)
Start concentrations are read from stdin (unless the -f option is used) in [mol/l], equilibrium
concentrations are given realtive to the sum of the inputs. An arbitrary number of initial
concentrations can be specified (one tuple of concentrations per line).
-f, --concfile=filename
Specify a file with initial concentrations for the input sequences.
The table consits of arbitrary many lines with multiple numbers separated by whitespace (the
concentration of the input sequences A, B, C, etc.). This option will automatically toggle the -c
(and thus -a and -p) options (see above).
--absolute-concentrations Report absolute instead of relative
concentrations
(default=off)
--betaScale=DOUBLE
Set the scaling of the Boltzmann factors. (default=`1.')
The argument provided with this option is used to scale the thermodynamic temperature in the
Boltzmann factors independently from the temperature of the individual loop energy contributions.
The Boltzmann factors then become 'exp(- dG/(kT*betaScale))' where 'k' is the Boltzmann constant,
'dG' the free energy contribution of the state and 'T' the absolute temperature.
-S, --pfScale=DOUBLE
In the calculation of the pf use scale*mfe as an estimate for the ensemble free energy (used to
avoid overflows).
(default=`1.07')
The default is 1.07, useful values are 1.0 to 1.2. Occasionally needed for long sequences.
--bppmThreshold=cutoff
Set the threshold/cutoff for base pair probabilities included in the postscript output.
(default=`1e-5')
By setting the threshold the base pair probabilities that are included in the output can be
varied. By default only those exceeding '1e-5' in probability will be shown as squares in the dot
plot. Changing the threshold to any other value allows for increase or decrease of data.
-g, --gquad
Incoorporate G-Quadruplex formation into the structure prediction algorithm.
(default=off)
Note, only intramolecular G-quadruplexes are considered.
Structure Constraints:
Command line options to interact with the structure constraints feature of this program
--maxBPspan=INT
Set the maximum base pair span.
(default=`-1')
--commands=filename
Read additional commands from file
Commands include hard and soft constraints, but also structure motifs in hairpin and internal
loops that need to be treeted differently. Furthermore, commands can be set for unstructured and
structured domains.
Energy Parameters:
Energy parameter sets can be adapted or loaded from user-provided input files
-T, --temp=DOUBLE
Rescale energy parameters to a temperature of temp C. Default is 37C.
(default=`37.0')
-P, --paramFile=paramfile
Read energy parameters from paramfile, instead of using the default parameter set.
Different sets of energy parameters for RNA and DNA should accompany your distribution. See the
RNAlib documentation for details on the file format. The placeholder file name 'DNA' can be used
to load DNA parameters without the need to actually specify any input file.
-4, --noTetra
Do not include special tabulated stabilizing energies for tri-, tetra- and hexaloop hairpins.
(default=off)
Mostly for testing.
--salt=DOUBLE
Set salt concentration in molar (M). Default is 1.021M.
Model Details:
Tweak the energy model and pairing rules additionally using the following parameters
-d, --dangles=INT
How to treat "dangling end" energies for bases adjacent to helices in free ends and multi-loops.
(default=`2')
With -d1 only unpaired bases can participate in at most one dangling end. With -d2 this check is
ignored, dangling energies will be added for the bases adjacent to a helix on both sides in any
case; this is the default for mfe and partition function folding (-p). The option -d0 ignores
dangling ends altogether (mostly for debugging). With -d3 mfe folding will allow coaxial stacking
of adjacent helices in multi-loops. At the moment the implementation will not allow coaxial
stacking of the two enclosed pairs in a loop of degree 3 and works only for mfe folding.
Note that with -d1 and -d3 only the MFE computations will be using this setting while partition
function uses -d2 setting, i.e. dangling ends will be treated differently.
--noLP Produce structures without lonely pairs (helices of length 1).
(default=off)
For partition function folding this only disallows pairs that can only occur isolated. Other pairs
may still occasionally occur as helices of length 1.
--noGU Do not allow GU pairs.
(default=off)
--noClosingGU
Do not allow GU pairs at the end of helices.
(default=off)
--nsp=STRING
Allow other pairs in addition to the usual AU,GC,and GU pairs.
Its argument is a comma separated list of additionally allowed pairs. If the first character is a
"-" then AB will imply that AB and BA are allowed pairs, e.g. --nsp="-GA" will allow GA and AG
pairs. Nonstandard pairs are given 0 stacking energy.
--energyModel=INT
Set energy model.
Rarely used option to fold sequences from the artificial ABCD... alphabet, where A pairs B, C-D
etc. Use the energy parameters for GC (--energyModel 1) or AU (--energyModel 2) pairs.
--helical-rise=FLOAT
Set the helical rise of the helix in units of Angstrom.
(default=`2.8')
Use with caution! This value will be re-set automatically to 3.4 in case DNA parameters are loaded
via -P DNA and no further value is provided.
--backbone-length=FLOAT
Set the average backbone length for looped regions in units of Angstrom.
(default=`6.0')
Use with caution! This value will be re-set automatically to 6.76 in case DNA parameters are
loaded via -P DNA and no further value is provided.
REFERENCES
If you use this program in your work you might want to cite:
R. Lorenz, S.H. Bernhart, C. Hoener zu Siederdissen, H. Tafer, C. Flamm, P.F. Stadler and I.L. Hofacker
(2011), "ViennaRNA Package 2.0", Algorithms for Molecular Biology: 6:26
I.L. Hofacker, W. Fontana, P.F. Stadler, S. Bonhoeffer, M. Tacker, P. Schuster (1994), "Fast Folding and
Comparison of RNA Secondary Structures", Monatshefte f. Chemie: 125, pp 167-188
R. Lorenz, I.L. Hofacker, P.F. Stadler (2016), "RNA folding with hard and soft constraints", Algorithms
for Molecular Biology 11:1 pp 1-13
The energy parameters are taken from:
D.H. Mathews, M.D. Disney, D. Matthew, J.L. Childs, S.J. Schroeder, J. Susan, M. Zuker, D.H. Turner
(2004), "Incorporating chemical modification constraints into a dynamic programming algorithm for
prediction of RNA secondary structure", Proc. Natl. Acad. Sci. USA: 101, pp 7287-7292
D.H Turner, D.H. Mathews (2009), "NNDB: The nearest neighbor parameter database for predicting stability
of nucleic acid secondary structure", Nucleic Acids Research: 38, pp 280-282
REPORTING BUGS
If in doubt our program is right, nature is at fault. Comments should be sent to rna@tbi.univie.ac.at.
RNAmultifold 2.7.0 September 2025 RNAMULTIFOLD(1)