Steps for using VADR for HAV annotation:
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Download and install the latest version of VADR, following the instructions on this page. Alternatively, you can use the StaPH-B VADR 1.6.4 docker image created by Curtis Kapsak (docker image names:
staphb/vadr:1.6.4andstaphb/vadr:latest), available on dockerhub and quay. A brief README for the docker image is here. -
Clone the latest HAV VADR model library from this repository.
git clone git@github.com:greninger-lab/vadr-models-hav.gitNote the path to the directory name created plus the "hav" subdirectory (e.g. /path/to/vadr-models-hav/hav) as
<hav-models-dir-path>for step 4. -
Remove terminal ambiguous nucleotides from your input fasta sequence file using the
fasta-trim-terminal-ambigs.plscript in$VADRSCRIPTSDIR/miniscripts/.To remove terminal ambiguous nucleotides from your sequence file
<input-fasta-file>and to remove short and long sequences to create a new trimmed file<trimmed-fasta-file>, execute:
$VADRSCRIPTSDIR/miniscripts/fasta-trim-terminal-ambigs.pl --minlen 50 --maxlen 8000 <input-fasta-file> > <trimmed-fasta-file>
- Run the
v-annotate.plprogram on an input trimmed fasta file with HAV sequences using the recommended command below.
v-annotate.pl -r --mkey hav --mdir <hav-models-dir-path> <fasta-file-to-annotate> <output-directory-to-create>
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After running the
v-annotate.plcommand in step 4, there will be a number of files generated in the<output-directory-to-create>. Among these files, there are 5-column tab-delimited feature table files that end with the suffix.tbl. There is a separate file for passing (XXXXX.vadr.pass.tbl) and failing (XXXXX.vadr.fail.tbl) sequences. The format of the.tblfiles is described here: https://www.ncbi.nlm.nih.gov/genbank/feature_table/More information about understanding failures and error alerts can be found in the VADR documentation here: https://github.com/ncbi/vadr/blob/master/documentation/annotate.md
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The VADR model library for HAV annotation was developed using representative sequences from the 6 genotypes currently available in the complete HAV genomes published in GenBank: OK625565(IA), KP879217(IB), AY644676(IIA), AY644670(IIB), DQ991029(IIIA) and AB258387(IIIB).
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Some of the model genomes have been modified slightly on the 3' end to have polyA tails of consistent length and facilitate consistent behavior across the models. These include:
- OK625565 (IA) removed 7 As from 3' end
- KP879217 (IB) added 18 As to 3' end
- DQ991029 (IIIA) removed 10 As from 3' end
- VADR README
- VADR installation instructions
v-build.plexample usage and command-line optionsv-annotate.plexample usage, command-line options and alert information- Explanations and examples of
v-annotate.pldetailed alert and error messages- Output fields with detailed alert and error messages
- Explanation of sequence and model coordinate fields in
.altfiles toy50toy model used in examples of alert messages- Examples of different alert types and corresponding
.altoutput - Posterior probability annotation in VADR output Stockholm alignments
- VADR output file formats
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The recommended citation for using VADR is: Alejandro A Schäffer, Eneida L Hatcher, Linda Yankie, Lara Shonkwiler, J Rodney Brister, Ilene Karsch-Mizrachi, Eric P Nawrocki; VADR: validation and annotation of virus sequence submissions to GenBank. BMC Bioinformatics 21, 211 (2020). https://doi.org/10.1186/s12859-020-3537-3
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This page was adapted for HAV from Mpox virus annotation