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v-scan.pl example usage and command-line options


Quickstart v-scan.pl examples

v-scan.pl can be run in 3 modes:

  1. model library is unknown but all sequences are expected to match to the same library, run like: v-scan.pl <fasta file to annotate> <output directory to create>. For example:

    v-scan.pl $VADRSCRIPTSDIR/documentation/scan-files/n5.fa vs-n5
    

    After running the output files will be in the newly created directory vs-n5.

  2. model libraries are unknown and sequences may match to different libraries, same as above but add the -m option:

    v-scan.pl -m $VADRSCRIPTSDIR/documentation/scan-files/m5.fa vs-m5
    
  3. model library is known and all sequences are expected to match to it, same as 1 but use the --only option:

    v-scan.pl --only norovirus $VADRSCRIPTSDIR/documentation/scan-files/n5.fa vs-n5-only
    

v-scan.pl config file

v-scan.pl can be used to annotate sequences that match to one or more VADR model libraries. v-scan.pl determines which model library to use for the input sequences and then calls v-annotate.pl to annotate all sequences that match to that model library. It will supply v-annotate.pl with the command-line options specific for that model library read from the input config file.

v-scan.pl requires a 'config' file that lists information on the model libraries it will use. The config file that will be used will be stored in the $VADRCONFIGFILE environment variable after the installation procedure. If you want you can modify this file or modify a copy of it, and specify that a different file <s> be used by using the -c <s> option to v-scan.pl or by changing the value of $VADRCONFIGFILE to point to a different file.

Here is the config file that is included with VADR and is used by default in vadr/vadr.config with comment lines removed for brevity (all lines that begin with a # are comment lines):

dengue    $VADRINSTALLDIR/vadr-models-flavi    --split --cpu 1 --group Dengue --nomisc --noprotid --mkey flavi -r
hcv       $VADRINSTALLDIR/vadr-models-flavi    --split --cpu 1 -r --mkey flavi --group HCV
flavi     $VADRINSTALLDIR/vadr-models-flavi    --split --cpu 1 -r --nomisc
norovirus $VADRINSTALLDIR/vadr-models-calici   --split --cpu 1 --group Norovirus --nomisc --noprotid --mkey calici -r
calici    $VADRINSTALLDIR/vadr-models-calici   --split --cpu 1 -r --nomisc 
sarscov2  $VADRINSTALLDIR/vadr-models-sarscov2 --split --cpu 4 -s -r --nomisc --lowsim5seq 6 --lowsim3seq 6 --alt_fail lowscore,insertnn,deletinn --glsearch
corona    $VADRINSTALLDIR/vadr-models-corona   --split --cpu 1 -s -r --nomisc --lowsim5seq 6 --lowsim3seq 6 --alt_fail lowscore,insertnn,deletinn --glsearch 
flu       $VADRINSTALLDIR/vadr-models-flu      --split --cpu 2 -r --atgonly --alt_fail extrant5,extrant3 --xnocomp --nomisc --forcegene
rsv       $VADRINSTALLDIR/vadr-models-rsv      --split --cpu 1 -r --xnocomp --nomisc 
mpxv      $VADRINSTALLDIR/vadr-models-mpxv     --split --cpu 1 --glsearch --minimap2 -s -r --nomisc --r_lowsimok --r_lowsimxd 100 --r_lowsimxl 2000 --alt_pass discontn,dupregin --s_overhang 150

(Each line prefixed with # is a comment line and is ignored by v-scan.pl.) All other lines have 3 or more fields:

idx field description
1 <options key> name for this library, a unique key that will be used for naming v-scan.pl output files, cannot contain whitespace
2 <model directory> path to the model directory that includes all model files for this library, the same model directory can be used for multiple <options key> values, cannot contain whitespace
3 to end <options string> the v-annotate.pl options that should be used for sequences matching this <options key> library during the annotation stage of v-scan.pl; this must contain --mkey <s> if the model files in the <model directory> are named with a key other than the <options key>, an example is the use of --mkey calici for the norovirus <options key> in the example file above; may contain whitespace

Note that in the above example config file, both dengue and hcv <options key> values use the flavi model library. We can tell this because --mkey flavi exists in the <options string> for the lines beginning with dengue and hcv, and because the <model directory> is the same for all three of dengue, hcv and flavi. Similarly norovirus uses the calici library.

Multiple <options string> values can use the same model libraries because sometimes we may want to use different v-annotate.pl options for different models within those libraries. In the example of dengue, hcv, and flavi, you can see that dengue uses the --noprotid and --group Dengue options, where as hcv and flavi use different options. All sequences that match to dengue will be annotated by v-annotate.pl using the dengue options, all sequences that match to hcv will be annotated using the hcv options, and all sequences that match to flavi will be annotated using the flavi options.

The way that sequences are matched to an <options key> is as follows: in the v-scan.pl classification stage, each sequence is scanned against each unique model library from the config file. In the example config file, this is only two model libraries:

  1. the flavi model library, with files named with the key flavi (e.g. flavi.minfo) in the directory $VADRINSTALLDIR/vadr-models-flavi.

  2. the calici model library, with files named with the key calici (e.g. calici.minfo) in the directory $VADRINSTALLDIR/vadr-models-calici.

The dengue and hcv <options key> values use the flavi library due to the --mkey flavi in their <options string>, and norovirus uses the calici library due to --mkey calici in its <options string>.

Then, when parsing the output for the the scan against the flavi library, sequences are matched to either dengue, hcv or flavi by checking if the best matching model for each sequence matches to dengue or hcv or flavi. A model matches to an <options key> if its model name, group or subgroup is identical to that <options key> after lowercasing and removing all special characters from the name, group or subgroup. For example, if a sequence's best matching model is NC_001477 which has group defined as Dengue in the flavi.minfo file (relevant line below)

MODEL NC_001477 blastdb:"NC_001477.vadr.protein.fa" group:"Dengue" length:"10735" subgroup:"1"

then that sequence will match to dengue and that sequence will then be annotated with v-annotate.pl using the dengue <options string>. Or, if a sequence matched to a model named HCV! then it would match to hcv because HCV! becomes hcv after making it lowercase and removing all special (non-alphanumeric) characters. Any sequence that matches best to a model in the flavi library that does not match either dengue or hcv in this way will match to flavi and v-annotate.pl will be used with the flavi options string to annotate it.

Similarly, when parsing the output for the scan against the calici library, sequences are matched to either norovirus or calici in the same way.

Model libraries do not have to be nested in this way. Each <options key> in the config file can pertain to its own unique model library.

Modifying the --split and --cpu <n> options

Note that in the default config file shown above, each <options string> includes the --split and --cpu <n> options with <n> varying between 1, 2 and 4. These options are used to parallelize processing in v-annotate.pl across <n> threads. If you prefer to run non-threaded, you can remove these two options from all the lines of the config file. You should also potentially change the number of threads that are used for each <options key> by modifying <n> after considering how much RAM you have on your computer The suggested amount of RAM per thread for each virus is:

options key recommended RAM per thread
sarscov2 2Gb
flu 4Gb
dengue, hcv, flavi, norovirus, calici, corona, mpxv 8Gb
rsv 16Gb

Adding libraries to the config file

You can download or build additional vadr model libraries and add them to the config file or make your own config file. To use a different config file <s> use the -c <s> option. The list of available VADR models and an example config file that uses them all is here.

Printing information from the config file

Several command-line options exist for outputting information from the config file, and on the models that are in the libraries listed in the config file: --l_all, --l_lib <s>, --l_dir, --l_opt and --l_mdl. Examples of these can be found below.


Walk-throughs of v-scan.pl examples

This section includes more detailed information on how to use v-scan.pl and understand its output.

To determine the command-line usage of v-scan.pl (or any VADR script), use the -h option, like this:

v-scan.pl -h 

You'll see something like the following output:

# v-scan.pl :: scan and annotate sequences against VADR model libraries 
# VADR 1.7 (Sep 2025)
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# date:    Tue Sep  2 10:39:19 2025
#
Usage: v-scan.pl [-options] <fasta file to annotate> <output directory to create>

The first few lines are the banner which show the name of the VADR script being run along with the version and release date. This is followed by the time and date the command was executed. The Usage: line details the expected command line arguments. v-scan.pl takes as input two command line arguments, a fasta file with sequences to analyze and annotate (<fasta file to annotate>) and the name of the output directory you want it to create (<output directory to create>) and populate with output files.

After that comes a list of all available command-line options. These are explained in more detail below.


Running v-scan.pl (mode 1)

Below is an example v-scan.pl command run in mode 1, where all sequences are expected to match to a single model library (using our default config file this would mean either 'flavi' or 'calici') but that library is unknown. For this example we will use the sequence file vadr/documentation/scan-files/n5.fa that includes 5 norovirus sequences, and we will create the directly vs-n5:

v-scan.pl $VADRSCRIPTSDIR/documentation/scan-files/n5.fa vs-n5

The standard output of v-scan.pl that is printed to the screen (which is also output to the .log output file) begins with the banner and date again followed by a list of relevant environment variables, the command line arguments used and any command line options used:

# date:             Tue Sep  2 14:16:40 2025
# $VADRCONFIGFILE:  /home/nawrocki/vadr-install-dir/vadr.config
# $VADRSCRIPTSDIR:  /home/nawrocki/vadr-install-dir
#
# sequence file:     /home/nawrocki/vadr-install-dir/documentation/scan-files/n5.fa
# output directory:  vs-n5

No command line options were used in our example output, but if they were information on them would have appeared after the output directory line.

Next, information is output about each step the script is proceeding through. When each step is completed, the elapsed time for that step is output.

v-scan.pl will use the default VADR config file ($VADRMODELDIR/default.vadr.config) to analyze the sequences in n5.fa, and will create an output directory named vs-5 and populate it with many output files.

The output of v-scan.pl lists the steps it takes:

# Validating input                                             ... done. [    0.0 seconds]
# Sampling 3 sequences to use for classification               ... done. [    0.0 seconds]
# Scanning 3 sequences against flavi  library                  ... done. [    3.3 seconds]
# Scanning 3 sequences against calici library                  ... done. [    3.8 seconds]

After validating the input, v-scan.pl samples 3 sequences to use for the classification stage. Only 3 sequences are used to make this stage more efficient based on the assumption is that all the sequences will be for the same model library. If v-scan.pl determines that more than one model library is matched by the sampled sequences, then it will fail with an error message indicating which libraries were matched. The --s_nseq <n> option will specify that <n> sequences be sampled instead of 3, and the --all option will turn off sampling and all sequences be used for classification. An example of allowing multiple libraries to match with the -m option is below.

After the classification stage, all of the sequences will be annotated using the matching library, which in this case is the norovirus library:

# Annotating 5 sequences with norovirus model library          ... done. [   31.7 seconds]
# Generating tabular output                                    ... done. [    0.0 seconds]
# Summary of sequences matching norovirus:
#
#                                      num   num   num
#idx  model      group      subgroup  seqs  pass  fail
#---  ---------  ---------  --------  ----  ----  ----
1     NC_039475  Norovirus  GII          2     2     0
2     NC_039477  Norovirus  GII          2     2     0
3     NC_039476  Norovirus  GII          1     1     0
#---  ---------  ---------  --------  ----  ----  ----
-     *all*      -          -            5     5     0
-     *none*     -          -            0     0     0
#---  ---------  ---------  --------  ----  ----  ----
#
# Zero alerts reported for seqs matching norovirus.

And finally the output files are listed:

# norovirus library output printed to screen saved in:                              vs-n5.norovirus.vadr.log
# norovirus library list of executed commands saved in:                             vs-n5.norovirus.vadr.cmd
# norovirus library list and description of all output files saved in:              vs-n5.norovirus.vadr.filelist
# norovirus library esl-seqstat -a output for input fasta file saved in:            vs-n5.norovirus.vadr.seqstat
# norovirus library 5 column feature table output for passing sequences saved in:   vs-n5.norovirus.vadr.pass.tbl
# norovirus library 5 column feature table output for failing sequences saved in:   vs-n5.norovirus.vadr.fail.tbl
# norovirus library list of passing sequences saved in:                             vs-n5.norovirus.vadr.pass.list
# norovirus library list of failing sequences saved in:                             vs-n5.norovirus.vadr.fail.list
# norovirus library list of alerts in the feature tables saved in:                  vs-n5.norovirus.vadr.alt.list
# norovirus library fasta file with passing sequences saved in:                     vs-n5.norovirus.vadr.pass.fa
# norovirus library fasta file with failing sequences saved in:                     vs-n5.norovirus.vadr.fail.fa
# norovirus library per-sequence tabular classification summary file saved in:      vs-n5.norovirus.vadr.sqc
# norovirus library per-model tabular summary file saved in:                        vs-n5.norovirus.vadr.mdl
# norovirus library per-sequence tabular annotation summary file saved in:          vs-n5.norovirus.vadr.sqa
# norovirus library per-feature tabular summary file saved in:                      vs-n5.norovirus.vadr.ftr
# norovirus library per-model-segment tabular summary file saved in:                vs-n5.norovirus.vadr.sgm
# norovirus library per-alert tabular summary file saved in:                        vs-n5.norovirus.vadr.alt
# norovirus library alert count tabular summary file saved in:                      vs-n5.norovirus.vadr.alc
# norovirus library alignment doctoring tabular summary file saved in:              vs-n5.norovirus.vadr.dcr
# norovirus library replaced stretches of Ns summary file (-r) saved in:            vs-n5.norovirus.vadr.rpn
#
# Output printed to screen saved in:                   vs-n5.vadr.log
# List of executed commands saved in:                  vs-n5.vadr.cmd
# List and description of all output files saved in:   vs-n5.vadr.filelist
# per-model library tabular summary file saved in:     vs-n5.vadr.lib
#
# All output files created in directory ./vs-n5/
#
# Elapsed time:  00:00:38.82
#                hh:mm:ss
# 
[ok]

All of these files were created in the newly created directory vs-n5. The files that include norovirus in their name were created by v-annotate.pl and more information on those files and their formats can be found here. The v-annotate.pl documentation also includes a description of several of these files.

The final four files are the .log file, which is the same as the standard output printed to the screen currently being discussed, the .cmd file, and the .filelist file which lists the output files created by v-scan.pl, and finally the .lib file which explains how many sequences matched to each library in the classification stage.


Running v-scan.pl (mode 2)

If there may be sequences that match to multiple libraries in the input fasta file, use the -m option, like this:

v-scan.pl -m $VADRSCRIPTSDIR/documentation/scan-files/m5.fa vs-m5

This will generate:

# Validating input                                             ... done. [    0.0 seconds]
# Scanning 5 sequences against flavi  library                  ... done. [    4.8 seconds]
# Scanning 5 sequences against calici library                  ... done. [    4.1 seconds]
# Annotating 1 dengue    sequences                             ... done. [   40.4 seconds]
# Annotating 1 flavi     sequences                             ... done. [   45.3 seconds]
# Annotating 2 norovirus sequences                             ... done. [   27.7 seconds]
# Annotating 1 calici    sequences                             ... done. [   17.3 seconds]
# Generating tabular output                                    ... done. [    0.0 seconds]
#
# Summary of sequences matching each library:
#
#     options    model    num
#idx  key        key     seqs
#---  ---------  ------  ----
1     dengue     flavi      1
2     hcv        flavi      0
3     flavi      flavi      1
4     norovirus  calici     2
5     calici     calici     1

You may notice a few differences between the output with -m. First, there is no sampling step, all sequences must be used for the classification step because with -m multiple libraries may be matched and we want to determine which library to use for annotating each sequence.

Also, following the summary of sequences matching each library, you will see per-library summary statistics for each of the four libraries matched, and a list of output sequences for each library.


Running v-scan.pl (mode 3)

If you know which library your sequences will match to, you can use the --only option, like this:

v-scan.pl --only norovirus $VADRSCRIPTSDIR/documentation/scan-files/n5.fa vs-n5-only
# Validating input                                             ... done. [    0.0 seconds]
# Annotating 5 sequences with norovirus model library          ... done. [   31.6 seconds]
#
# Summary of sequences matching norovirus:
#
#                                      num   num   num
#idx  model      group      subgroup  seqs  pass  fail
#---  ---------  ---------  --------  ----  ----  ----
1     NC_039475  Norovirus  GII          2     2     0
2     NC_039477  Norovirus  GII          2     2     0
3     NC_039476  Norovirus  GII          1     1     0
#---  ---------  ---------  --------  ----  ----  ----
-     *all*      -          -            5     5     0
-     *none*     -          -            0     0     0
#---  ---------  ---------  --------  ----  ----  ----

Note that the sampling step and classification step performed in mode 1 are skipped. This makes it slightly more efficient to use --only if you know the library.

If you want to restrict the libraries that can be matched, but not specify a single one, you can use --only with a list of comma separated library keys, like this: v-scan.pl --only norovirus,flavi n5.fa vs-n5-only2. Alternatively, you can list libraries that you want v-scan.pl to skip (ignore) using the --skip option like this: v-scan.pl --skip dengue,flavi n5.fa vs-n5-skip.


v-scan.pl command-line options

To get a list of command-line options, execute:

v-scan.pl -h

This will output the usage and available command-line options. Each option has a short description, but additional information on some of these options can be found below. For v-scan.pl the available options are split into four different categories, each explained in their own subsection below.

In the tables describing options below, <s> represents a string, <x> indicates a floating point number and <n> represents an integer.

v-scan.pl basic options

......option.... explanation
-f if <output directory> already exists, then using this option will cause it to be overwritten, otherwise the progam exits in error
-m multiple-library mode, allow matches to multiple model libraries, without this option matches to only one library are allowed and the program will exit if matches to multiple libraries are found
-c <s> use the config file <s> instead of the config file in $VADRCONFIGFILE
-v verbose mode: all commands will be output to standard output as they are run
--first specify that if a sequence matches to more than one library, use the first one; by default the higher scoring match is used
--cpu <n> parallelize classification stage across CPU workers, requires -m, only impacts the initial classification stage, parallelization of annotation stage can be controlled by adding --split --cpu <n> to options strings in the config file
--lone exit if at least one sequence matches to multiple libraries
--origfa do not copy the input fasta file into output directory prior to analysis, use the original
--keep keep additional v-annotate.pl output files that are normally removed

v-scan.pl options for specifying which model libraries to use

..........option.......... explanation
--only <s> only use the model library(ies) with option keys (e.g. flavi) listed in the comma separated string <s>, all option keys must exist in config file
--skip <s> do not use the model library(ies) with option keys (e.g. flavi) listed in the comma separated string <s>, all option keys must exist in config file

v-scan.pl options related to the random sampling of sequences for determining model library to use (sampling is turned off if -m is used or only one model library is being used)

............option............ explanation
--all do not sample, pick model library(ies) based on all sequences (automatically turned on if -m used)
--s_nseq <n> set the number of sequences to sample to <n>, default value is 3
--s_beg sample sequences from the beginning of the sequence file, not randomly
--s_seed <n> set the random number generator seed to <n>, default value is 181

v-scan.pl options for listing information from the config file or about models and exiting

............option............ explanation
--l_all list information about all models, model directories, and options strings in the config file and exit
--l_lib <s> list all information about the model library for options key <s> (e.g. flavi) in the config file and exit
--l_dir list all model directories in the config file and exit
--l_opt list v-annotate.pl options for each option key in the config file and exit
--l_mdl list information about all the models in all libraries in the config file and exit

Using the list options

The v-scan.pl options listed above beginning with -l can be useful for listing information about the config file and the model libraries listed in the config file. Here are some examples of using these options when the environment variable $VADRCONFIGFILE points to the default config file in $VADRINSTALLDIR/default.vadr.config, which it should be default after following the installation instructions:


List the model directories in the config file, for all libraries:

v-scan.pl --l_dir
############################################################
#
# VADR 1.7 (Sep 2025)
#
# config file: /home/nawrocki/vadr-install-dir/default.vadr.config
#
# Model library directory information:
#
#options key  model key  model dir
#-----------  ---------  ---------
dengue        flavi      /home/nawrocki/vadr-install-dir/vadr-models-flavi
hcv           flavi      /home/nawrocki/vadr-install-dir/vadr-models-flavi
flavi         "          /home/nawrocki/vadr-install-dir/vadr-models-flavi
norovirus     calici     /home/nawrocki/vadr-install-dir/vadr-models-calici
calici        "          /home/nawrocki/vadr-install-dir/vadr-models-calici
sarscov2      "          /home/nawrocki/vadr-install-dir/vadr-models-sarscov2
corona        "          /home/nawrocki/vadr-install-dir/vadr-models-corona
flu           "          /home/nawrocki/vadr-install-dir/vadr-models-flu
rsv           "          /home/nawrocki/vadr-install-dir/vadr-models-rsv
mpxv          "          /home/nawrocki/vadr-install-dir/vadr-models-mpxv
#

#

List the v-annotate.pl options in the config file, for all libraries:

v-scan.pl --l_opt
# Options information:
#
#options key  v-annotate.pl options
#-----------  ---------------------
dengue        --split --cpu 1 --group Dengue --nomisc --noprotid --mkey flavi -r
hcv           --split --cpu 1 -r --mkey flavi --group HCV
flavi         --split --cpu 1 -r --nomisc
norovirus     --split --cpu 1 --group Norovirus --nomisc --noprotid --mkey calici -r
calici        --split --cpu 1 -r --nomisc
sarscov2      --split --cpu 4 -s -r --nomisc --lowsim5seq 6 --lowsim3seq 6 --alt_fail lowscore,insertnn,deletinn --glsearch
corona        --split --cpu 1 -s -r --nomisc --lowsim5seq 6 --lowsim3seq 6 --alt_fail lowscore,insertnn,deletinn --glsearch
flu           --split --cpu 2 -r --atgonly --alt_fail extrant5,extrant3 --xnocomp --nomisc --forcegene
rsv           --split --cpu 1 -r --xnocomp --nomisc
mpxv          --split --cpu 1 --glsearch --minimap2 -s -r --nomisc --r_lowsimok --r_lowsimxd 100 --r_lowsimxl 2000 --alt_pass discontn,dupregin --s_overhang 150
#

List all models in the model libraries in the config file:

v-scan.pl --l_mdl
# List of models in each library:
#
#idx   model key  options key  model name  length  group          subgroup
#----  ---------  -----------  ----------  ------  -------------  --------
1.1    flavi      dengue       NC_001477    10735  Dengue         1       
1.2    flavi      dengue       NC_001474    10723  Dengue         2       
1.3    flavi      dengue       NC_001475    10707  Dengue         3       
1.4    flavi      dengue       NC_002640    10649  Dengue         4       
1.5    flavi      hcv          NC_004102     9646  HCV            1       
1.6    flavi      hcv          NC_038882     9599  HCV            1       
1.7    flavi      hcv          NC_009823     9711  HCV            2       
1.8    flavi      hcv          NC_009824     9456  HCV            3       
1.9    flavi      hcv          NC_009825     9355  HCV            4       
1.10   flavi      hcv          NC_009826     9343  HCV            5       
1.11   flavi      hcv          NC_009827     9628  HCV            6       
1.12   flavi      hcv          NC_030791     9443  HCV            7       
1.13   flavi      "            NC_040815     8684  -              -       
1.14   flavi      "            NC_040788    10311  -              -       
1.15   flavi      "            NC_040776    10794  -              -       
..snip..
1.153  flavi      "            NC_001837     9550  -              -       
1.154  flavi      "            NC_001710     9392  -              -       
1.155  flavi      "            NC_001461    12573  -              -       
1.156  flavi      "            NC_031327    10588  -              -       
#----  ---------  -----------  ----------  ------  -------------  --------
2.1    calici     norovirus    NC_001959     7654  Norovirus      GI      
2.2    calici     norovirus    NC_008311     7382  Norovirus      GV      
2.3    calici     norovirus    NC_029645     7313  Norovirus      GIII    
2.4    calici     norovirus    NC_029646     7518  Norovirus      GII     
..snip..
5.67   flu        "            CY125947      1426  fluA-seg6      N11     
5.68   flu        "            CY125948      1027  fluA-seg7      -       
5.69   flu        "            CY125949       895  fluA-seg8      -       
5.70   flu        "            ON637239      1686  fluA-seg4      H19     
#----  ---------  -----------  ----------  ------  -------------  --------
6.1    rsv        "            KY654518     15277  RSV            A       
6.2    rsv        "            MZ516105     15276  RSV            B       
#----  ---------  -----------  ----------  ------  -------------  --------
7.1    mpxv       "            NC_063383   197209  Orthopoxvirus  Monkeypox_virus

List all information for a particular library/options key:

v-scan.pl --l_lib norovirus

This will print all the above information (model directory, options, model information), but only for norovirus.


List all information for all library/options keys:

v-scan.pl --l_all

This will print all the above information (model directory, options, model information), for all libraries.


Questions, comments or feature requests? Send a mail to eric.nawrocki@nih.gov.