the opera of phantome - 2017 (presented at the 22nd biennial evergreen phage meeting)

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The Opera of PhAnToMe 2017 Ramy K. Aziz (@azizrk) Aug 6, 2017 opus (LT) = work (Pl. opera) SEED-based phage database (2009-2013-…) Phage Genomics Workshop, Evergreen 2017 giantmicrobes.com

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Page 1: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

The Opera of PhAnToMe 2017

Ramy K. Aziz (@azizrk)

Aug 6, 2017

opus (LT) = work (Pl. opera)

SEED-based phage database (2009-2013-…)

Phage Genomics Workshop, Evergreen 2017

giantmicrobes.com

Page 2: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

6 August 2017

History

Phage Genomics - Evergreen 2017

NSF-funded, 3-year project (09-

12) to develop

Phage

Annotation

Tools and

Methods

Four Centers:

- SDSU, San Diego, CA

- VCU, Richmond, VA

- USF, St. Pete FL

- UA, Tucson, AZ

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http://www.phantome.org

6 August 2017

Web portal

Phage Genomics - Evergreen 2017

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PhAnToMe ToolBox

http://www.phantome.org

6 August 2017 Phage Genomics - Evergreen 2017

Very useful

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The ToolBox: The RAST family

• (At least) Five ways to annotate a genome via RAST:

– RAST (http://rast.nmpdr.org)

• annotates online, saves your genome on server

– myRAST (local)

• uses the server but you can edit offline)

– “PhAST” (http://www.phantome.org/PhageSeed/Phage.cgi?page=phast)

• phage-optimized gene-calling

– Use your favorite gene caller then upload gbk file to RAST

– RASTtk (second-generation RAST)

• modular

• batch upload

6 August 2017 Phage Genomics - Evergreen 2017

Page 6: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

http://rast.nmpdr.org

6 August 2017 Phage Genomics - Evergreen 2017

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http://rast.nmpdr.org

6 August 2017 Phage Genomics - Evergreen 2017

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“PhAST”: phage-optimized RAST

6 August 2017 Phage Genomics - Evergreen 2017

http://www.phantome.org/PhageSeed/Phage.cgi?page=phast

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“PhAST”: phage-optimized RAST

6 August 2017 Phage Genomics - Evergreen 2017

http://www.phantome.org/PhageSeed/Phage.cgi?page=phast

Page 10: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

RASTtk (RAST toolkit)

6 August 2017 Phage Genomics - Evergreen 2017

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RASTtk command-line

6 August 2017 Phage Genomics - Evergreen 2017

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RAST Video demos available

• Find & watch:

– http://tutorial.theseed.org

6 August 2017 Phage Genomics - Evergreen 2017

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PATRIC

6 August 2017 Phage Genomics - Evergreen 2017

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PATRIC

6 August 2017 Phage Genomics - Evergreen 2017

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PATRIC

6 August 2017 Phage Genomics - Evergreen 2017

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DEMO

How to?

Phage Genomics - Evergreen 2017

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What do you need

to annotate your genome?

• A sequenced genome

• Format: fasta or genbank (.gbk)

• A RAST username and password

• You can find some data to play with at

http://egybio.net/tutorial/

6 August 2017 Phage Genomics - Evergreen 2017

Page 18: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

1. Optimize RASTtk for phage

6 August 2017 Phage Genomics - Evergreen 2017

Page 19: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

1. Optimize RASTtk for phage

6 August 2017 Phage Genomics - Evergreen 2017

Page 20: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

1. Optimize RASTtk for phage

6 August 2017 Phage Genomics - Evergreen 2017

Page 21: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

1. Optimize RASTtk for phage

6 August 2017 Phage Genomics - Evergreen 2017

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1. Optimize RASTtk for phage

6 August 2017 Phage Genomics - Evergreen 2017

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1. Optimize RASTtk for phage

6 August 2017 Phage Genomics - Evergreen 2017

?

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1. Optimize RASTtk for phage

6 August 2017 Phage Genomics - Evergreen 2017

?

Page 25: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

1. Optimize RASTtk for phage

6 August 2017 Phage Genomics - Evergreen 2017

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1. Optimize RASTtk for phage

6 August 2017 Phage Genomics - Evergreen 2017

x

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1. Optimize RASTtk for phage

6 August 2017 Phage Genomics - Evergreen 2017

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2. Command-line RASTtk (Batch option)

• Where?

– On IRIS

(http://tutorial.theseed.org/services/docs/invoc

ation/Iris/iris.html)

– On your desktop (Download RASTtk)

6 August 2017 Phage Genomics - Evergreen 2017

Page 29: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

2. Command-line RASTtk (Batch option)

• How?

– The key is to convert each contigs file (fasta)

to so-called “Genome-typed object” GTO

– Once you have GTOs for all your genomes,

you can run a couple of commands to

annotate each of them OR put them in a

folder and annotate in batch

6 August 2017 Phage Genomics - Evergreen 2017

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2. Command-line RASTtk (Batch option)

• Commands:

o rast-create-genome

rast-create-genome --scientific-name ”Enterophage

Lambda" --genetic-code 11 --domain Virus --contigs

lambda.fasta > lambda.gto

o rast-process-genome

o rast-process-genome-batch

o rast-export-genome

Details on http://tutorial.theseed.org or follow

link from: http://egybio.net/tutorial

6 August 2017 Phage Genomics - Evergreen 2017

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3. Browse your favorite genome

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3. Browse your favorite genome

6 August 2017 Phage Genomics - Evergreen 2017

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4. Explore the protein page

• Annotation history

• Compare region

• Evidence

– similarities

– literature

6 August 2017 Phage Genomics - Evergreen 2017

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4. Explore the protein page

6 August 2017 Phage Genomics - Evergreen 2017

• Find your favorite protein

Page 35: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

4. Explore the protein page

6 August 2017 Phage Genomics - Evergreen 2017

• Find your favorite protein

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5. Align proteins (in context)

• Evidence> Similarities> Align

• Compare region, advanced settings

• Phylogenetic trees

6 August 2017 Phage Genomics - Evergreen 2017

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5. Align proteins (in context)

6 August 2017 Phage Genomics - Evergreen 2017

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THE COMMUNITY

The Opera of PhAnToMe 2.0

6 August 2017 Phage Genomics - Evergreen 2017

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SEED allows community annotation

6 August 2017 Phage Genomics - Evergreen 2017

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Annotations will improve

only if YOU help

6 August 2017 Phage Genomics - Evergreen 2017

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CREDITS

The Opera of PhAnToMe 2.0

6 August 2017 Phage Genomics - Evergreen 2017

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Acknowledgments

Robert A. Edwards, PhD

• RASTtk and PhiRAST development:

Ross Overbeek, Robert Olson, Jim Davis, Gordon

Pusch, Terry Disz, Bruce Parrello

• Phage annotators (Phantomers):

Bhakti Dwivedi, Mya Breitbart, et al.

• FIG and all SEED annotators:

VeronikaV, SvetaG, OlgaV/Z, et al.

6 August 2017

$$

Phage Genomics - Evergreen 2017

&

NSF

Katelyn McNair

Page 43: The Opera of Phantome - 2017 (presented at the 22nd Biennial Evergreen Phage Meeting)

If you use, please cite

• SEED, RAST, myRAST, phiRAST, PHAST:

– RAST: Aziz et al., BMC Genomics 2008

– SEED servers: Aziz RK,, et al. (2012) PLoS ONE 7(10): e48053.

– Nucleic Acids Res. 2014 Jan;42(Database issue):D206-14

– PATRIC: Antonopolus et al., Brief. Bioinf. 2017 Jul 31; Online

early

6 August 2017 Phage Genomics - Evergreen 2017

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Questions?