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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
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
http://www.phantome.org
6 August 2017
Web portal
Phage Genomics - Evergreen 2017
PhAnToMe ToolBox
http://www.phantome.org
6 August 2017 Phage Genomics - Evergreen 2017
Very useful
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
http://rast.nmpdr.org
6 August 2017 Phage Genomics - Evergreen 2017
http://rast.nmpdr.org
6 August 2017 Phage Genomics - Evergreen 2017
“PhAST”: phage-optimized RAST
6 August 2017 Phage Genomics - Evergreen 2017
http://www.phantome.org/PhageSeed/Phage.cgi?page=phast
“PhAST”: phage-optimized RAST
6 August 2017 Phage Genomics - Evergreen 2017
http://www.phantome.org/PhageSeed/Phage.cgi?page=phast
RASTtk (RAST toolkit)
6 August 2017 Phage Genomics - Evergreen 2017
RASTtk command-line
6 August 2017 Phage Genomics - Evergreen 2017
RAST Video demos available
• Find & watch:
– http://tutorial.theseed.org
6 August 2017 Phage Genomics - Evergreen 2017
PATRIC
6 August 2017 Phage Genomics - Evergreen 2017
PATRIC
6 August 2017 Phage Genomics - Evergreen 2017
PATRIC
6 August 2017 Phage Genomics - Evergreen 2017
DEMO
How to?
Phage Genomics - Evergreen 2017
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
1. Optimize RASTtk for phage
6 August 2017 Phage Genomics - Evergreen 2017
1. Optimize RASTtk for phage
6 August 2017 Phage Genomics - Evergreen 2017
1. Optimize RASTtk for phage
6 August 2017 Phage Genomics - Evergreen 2017
✔
1. Optimize RASTtk for phage
6 August 2017 Phage Genomics - Evergreen 2017
✔
1. Optimize RASTtk for phage
6 August 2017 Phage Genomics - Evergreen 2017
1. Optimize RASTtk for phage
6 August 2017 Phage Genomics - Evergreen 2017
?
1. Optimize RASTtk for phage
6 August 2017 Phage Genomics - Evergreen 2017
?
1. Optimize RASTtk for phage
6 August 2017 Phage Genomics - Evergreen 2017
1. Optimize RASTtk for phage
6 August 2017 Phage Genomics - Evergreen 2017
✔
x
1. Optimize RASTtk for phage
6 August 2017 Phage Genomics - Evergreen 2017
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
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
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
3. Browse your favorite genome
6 August 2017 Phage Genomics - Evergreen 2017
3. Browse your favorite genome
6 August 2017 Phage Genomics - Evergreen 2017
4. Explore the protein page
• Annotation history
• Compare region
• Evidence
– similarities
– literature
6 August 2017 Phage Genomics - Evergreen 2017
4. Explore the protein page
6 August 2017 Phage Genomics - Evergreen 2017
• Find your favorite protein
4. Explore the protein page
6 August 2017 Phage Genomics - Evergreen 2017
• Find your favorite protein
5. Align proteins (in context)
• Evidence> Similarities> Align
• Compare region, advanced settings
• Phylogenetic trees
6 August 2017 Phage Genomics - Evergreen 2017
5. Align proteins (in context)
6 August 2017 Phage Genomics - Evergreen 2017
THE COMMUNITY
The Opera of PhAnToMe 2.0
6 August 2017 Phage Genomics - Evergreen 2017
SEED allows community annotation
6 August 2017 Phage Genomics - Evergreen 2017
Annotations will improve
only if YOU help
6 August 2017 Phage Genomics - Evergreen 2017
CREDITS
The Opera of PhAnToMe 2.0
6 August 2017 Phage Genomics - Evergreen 2017
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
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
Questions?

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

  • 1. 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
  • 2. 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
  • 3. http://www.phantome.org 6 August 2017 Web portal Phage Genomics - Evergreen 2017
  • 4. PhAnToMe ToolBox http://www.phantome.org 6 August 2017 Phage Genomics - Evergreen 2017 Very useful
  • 5. 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
  • 6. http://rast.nmpdr.org 6 August 2017 Phage Genomics - Evergreen 2017
  • 7. http://rast.nmpdr.org 6 August 2017 Phage Genomics - Evergreen 2017
  • 8. “PhAST”: phage-optimized RAST 6 August 2017 Phage Genomics - Evergreen 2017 http://www.phantome.org/PhageSeed/Phage.cgi?page=phast
  • 9. “PhAST”: phage-optimized RAST 6 August 2017 Phage Genomics - Evergreen 2017 http://www.phantome.org/PhageSeed/Phage.cgi?page=phast
  • 10. RASTtk (RAST toolkit) 6 August 2017 Phage Genomics - Evergreen 2017
  • 11. RASTtk command-line 6 August 2017 Phage Genomics - Evergreen 2017
  • 12. RAST Video demos available • Find & watch: – http://tutorial.theseed.org 6 August 2017 Phage Genomics - Evergreen 2017
  • 13. PATRIC 6 August 2017 Phage Genomics - Evergreen 2017
  • 14. PATRIC 6 August 2017 Phage Genomics - Evergreen 2017
  • 15. PATRIC 6 August 2017 Phage Genomics - Evergreen 2017
  • 16. DEMO How to? Phage Genomics - Evergreen 2017
  • 17. 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
  • 18. 1. Optimize RASTtk for phage 6 August 2017 Phage Genomics - Evergreen 2017
  • 19. 1. Optimize RASTtk for phage 6 August 2017 Phage Genomics - Evergreen 2017
  • 20. 1. Optimize RASTtk for phage 6 August 2017 Phage Genomics - Evergreen 2017 ✔
  • 21. 1. Optimize RASTtk for phage 6 August 2017 Phage Genomics - Evergreen 2017 ✔
  • 22. 1. Optimize RASTtk for phage 6 August 2017 Phage Genomics - Evergreen 2017
  • 23. 1. Optimize RASTtk for phage 6 August 2017 Phage Genomics - Evergreen 2017 ?
  • 24. 1. Optimize RASTtk for phage 6 August 2017 Phage Genomics - Evergreen 2017 ?
  • 25. 1. Optimize RASTtk for phage 6 August 2017 Phage Genomics - Evergreen 2017
  • 26. 1. Optimize RASTtk for phage 6 August 2017 Phage Genomics - Evergreen 2017 ✔ x
  • 27. 1. Optimize RASTtk for phage 6 August 2017 Phage Genomics - Evergreen 2017
  • 28. 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
  • 29. 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
  • 30. 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
  • 31. 3. Browse your favorite genome 6 August 2017 Phage Genomics - Evergreen 2017
  • 32. 3. Browse your favorite genome 6 August 2017 Phage Genomics - Evergreen 2017
  • 33. 4. Explore the protein page • Annotation history • Compare region • Evidence – similarities – literature 6 August 2017 Phage Genomics - Evergreen 2017
  • 34. 4. Explore the protein page 6 August 2017 Phage Genomics - Evergreen 2017 • Find your favorite protein
  • 35. 4. Explore the protein page 6 August 2017 Phage Genomics - Evergreen 2017 • Find your favorite protein
  • 36. 5. Align proteins (in context) • Evidence> Similarities> Align • Compare region, advanced settings • Phylogenetic trees 6 August 2017 Phage Genomics - Evergreen 2017
  • 37. 5. Align proteins (in context) 6 August 2017 Phage Genomics - Evergreen 2017
  • 38. THE COMMUNITY The Opera of PhAnToMe 2.0 6 August 2017 Phage Genomics - Evergreen 2017
  • 39. SEED allows community annotation 6 August 2017 Phage Genomics - Evergreen 2017
  • 40. Annotations will improve only if YOU help 6 August 2017 Phage Genomics - Evergreen 2017
  • 41. CREDITS The Opera of PhAnToMe 2.0 6 August 2017 Phage Genomics - Evergreen 2017
  • 42. 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
  • 43. 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