# Formatting annotated genomes for submission to NCBI

**URL:** <https://help.galaxyproject.org/t/formatting-annotated-genomes-for-submission-to-ncbi/17716>\
**Category:** usegalaxy.eu support\
**Tags:** troubleshooting, ncbi-submission\
**Created:** [May 9, 2026, 5:20am UTC](https://help.galaxyproject.org/t/formatting-annotated-genomes-for-submission-to-ncbi/17716 "2026-05-09T05:20:52Z")\
**Posts on this page:** 7\
**Page:** 1

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**Author:** ![jaredbernard](https://sea2.discourse-cdn.com/flex020/user_avatar/help.galaxyproject.org/jaredbernard/32/2595_2.png) [@jaredbernard](https://help.galaxyproject.org/u/jaredbernard)\
**Post date:** [May 9, 2026, 5:20am UTC](https://help.galaxyproject.org/t/formatting-annotated-genomes-for-submission-to-ncbi/17716/1 "2026-05-09T05:20:52Z")

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This topic may be a little outside of the ones normally on Galaxy Help – sorry, but I have been scouring the web and searched BioStars with no luck. I’ve just spent a number of years assembling, annotating, and analyzing genomes. Now I’m battling table2asn to produce sqn files fit for submission to NCBI. Of course, there are errors, but no clear way of dealing with the errors, so any ideas will be more than welcomed:

- Does Galaxy Europe have a converter tool for producing sqn files, either table2asn (converting gff3 + fasta to sequin) or GB2sequin (converting genbank to sequin)? I’ve tried looking around without any luck.
- My gff3 files have “Ontology\_term” and the sqn files want “product”, so all my carefully added ontology goes out the window and all the thousands of genes are called “hypothetical protein”. I thought maybe funannotate could help fix a gff3 that’s already annotated but I don’t yet see if this is possible. Also, my TE annotations have “classification”.
- There are errors such as SEQ\_FEAT.Range, etc. You can find a list of what they mean [here](https://www.ncbi.nlm.nih.gov/genbank/genome_validation/), but the point is that there should be a tool that fixes them. The -c f tag in table2sqn does not seem to, as far as I can tell. Is there a tool to help with this?

Thanks for any ideas!

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**Author:** ![jennaj](https://sea2.discourse-cdn.com/flex020/user_avatar/help.galaxyproject.org/jennaj/32/27_2.png) [@jennaj](https://help.galaxyproject.org/u/jennaj)\
**Post date:** [May 11, 2026, 8:07pm UTC](https://help.galaxyproject.org/t/formatting-annotated-genomes-for-submission-to-ncbi/17716/2 "2026-05-11T20:07:13Z")

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HI @jaredbernard

Yes, the default submission is created by the standardized processes. This is different from curation activities (which it sounds like you have been working on!).

Galaxy doesn’t have a replacement for the recommendations from NCBI here → [Prokaryotic and Eukaryotic Genomes Submission Guide](https://www.ncbi.nlm.nih.gov/genbank/genomesubmit/). Or, at least not that I know of! I shared your question with some others to see what they think, so you may get another reply!

If you want to share back whether you are working on prokaryotes or eukaryotes that might be helpful. And, I think you can get help with prokaryote annotation from NCBI but please double check me at the guide above!

You could also explore a tool like this one. It is **experimental only** (so purely for exploratory reasons!), but perhaps comparing the content between these output files (run on a small sample you have) helps to understand the expected file content for a submission?

- **NCBI EGAPx** annotates eukaryotic genomes

Let’s start there, thanks! 🙂

* * *

**Xref**

- 🎓 GTN [Genome Annotation / Tutorial List](https://galaxyproject.github.io/training-material/topics/genome-annotation/)
- ⚙ IWC [Intergalactic Workflow Commission](https://iwc.galaxyproject.org/?filter=Genome+Annotation) (filter=Genome+Annotation)

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<div class="post-metadata">

**Author:** ![jaredbernard](https://sea2.discourse-cdn.com/flex020/user_avatar/help.galaxyproject.org/jaredbernard/32/2595_2.png) [@jaredbernard](https://help.galaxyproject.org/u/jaredbernard)\
**Post date:** [May 11, 2026, 10:15pm UTC](https://help.galaxyproject.org/t/formatting-annotated-genomes-for-submission-to-ncbi/17716/3 "2026-05-11T22:15:30Z")

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My genomes are for eukaryotes, but they are already annotated. I don’t want to start over on annotation and I don’t want to submit my genomes without annotation after all that work. The pipeline I used was standard: Maker, EDTA, Blast2GO. Yet it seems that my gff3s are not formatted in a way that works with table2asn. As I said, my gff3s have Ontology\_terms whereas the NCBI sqn files require “products”. So I need tools to help with formatting, but haven’t found anything yet. Thanks for giving it some thought.

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**Author:** ![jennaj](https://sea2.discourse-cdn.com/flex020/user_avatar/help.galaxyproject.org/jennaj/32/27_2.png) [@jennaj](https://help.galaxyproject.org/u/jennaj)\
**Post date:** [May 13, 2026, 9:42pm UTC](https://help.galaxyproject.org/t/formatting-annotated-genomes-for-submission-to-ncbi/17716/4 "2026-05-13T21:42:49Z")

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Hi @jaredbernard

Thanks for explaining more! From here, I would suggest writing into NCBI to get clarification? To learn how they would prefer the curated ontology annotation to be handled?

Your question will be about how to supply additional GFF3 attributes and to confirm which base attributes are required, or dependencies between attributes, or what the minimum set is.

- You have `Ontology_term=NNNN`.
- But probably also need: `gene` and `CDS`, and possibly `mRNA`, `exon`.

The term “product” is defined here under FEATURE TABLE FORMAT

- [What is table2asn?](https://www.ncbi.nlm.nih.gov/genbank/table2asn/) → [https://ftp.ncbi.nlm.nih.gov/asn1-converters/by\_program/table2asn/DOCUMENTATION/table2asn\_readme.txt](https://ftp.ncbi.nlm.nih.gov/asn1-converters/by_program/table2asn/DOCUMENTATION/table2asn_readme.txt)

```auto
The feature table specifies the location and type of biological features. table2asn will process the feature intervals and translate coding region (CDS) features into proteins.

The first and second columns are the start and stop locations of the feature, respectively, the third column is the type of feature (the feature key, e.g., gene, mRNA, CDS), the fourth column is a qualifier name (e.g., "product", and the fifth is a qualifier value (e.g., the name of the protein or gene). A simple example is:

  >Feature sde3n
  240 4084 gene
                          gene SDE3
  240 1361 mRNA
  1450 1641
  1730 3184
  3275 4084
                          product RNA helicase SDE3
  579 1361 CDS
  1450 1641
  1730 3184
  3275 3880
                          product RNA helicase SDE3

```

```auto
A simple example of a gene that is on the minus strand and is partial at its 3' end is:

  >Feature eno3
  1018 >1 gene
                          gene ENO3
  1018 >1 CDS
                          product beta-enolase

```

Be sure to review how ontology [terms are used](https://www.insdc.org/submitting-standards/feature-table/). These are called out like this in the readme above. This is different from `Ontology_terms` and if that was used in the tabular input, the values are in the 4th column and might explain your error. Or, maybe you need to add in the `product` line first (name of the protein)?

```auto
Gene Ontology (GO) terms can be indicated with the following qualifiers:

                          go_component endoplasmic reticulum|0005783
                          go_process glycolysis and gluconeogenesis|57|89197757|ACT,TEM
                          go_function excision repair|93||IPD

```

I see your same question over at Biostars. I’m going to link it in case you do get a reply, and someone else searches here in the future about the same question. → [https://www.biostars.org/p/9619529/](https://www.biostars.org/p/9619529/)

It is hard to guess more but I hope this helps! 🙂

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<div class="post-metadata">

**Author:** ![jaredbernard](https://sea2.discourse-cdn.com/flex020/user_avatar/help.galaxyproject.org/jaredbernard/32/2595_2.png) [@jaredbernard](https://help.galaxyproject.org/u/jaredbernard)\
**Post date:** [May 16, 2026, 11:18pm UTC](https://help.galaxyproject.org/t/formatting-annotated-genomes-for-submission-to-ncbi/17716/5 "2026-05-16T23:18:53Z")

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Thank you for these helpful tips, @jennaj! I also got some potentially good advice on Biostars too. I will see if I can get these steps to work. I did try corresponding with NCBI as you suggested and didn’t get a reply, but I will let you know how this goes. In the meantime, if any other tools are useful, I’m still very interested.

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<div class="post-metadata">

**Author:** ![jennaj](https://sea2.discourse-cdn.com/flex020/user_avatar/help.galaxyproject.org/jennaj/32/27_2.png) [@jennaj](https://help.galaxyproject.org/u/jennaj)\
**Post date:** [May 17, 2026, 5:56am UTC](https://help.galaxyproject.org/t/formatting-annotated-genomes-for-submission-to-ncbi/17716/6 "2026-05-17T05:56:20Z")

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Great, it looks like they noticed similar items and offered some help to solve them! Hope this works out! 🙂

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<div class="post-metadata">

**Author:** ![jaredbernard](https://sea2.discourse-cdn.com/flex020/user_avatar/help.galaxyproject.org/jaredbernard/32/2595_2.png) [@jaredbernard](https://help.galaxyproject.org/u/jaredbernard)\
**Post date:** [June 16, 2026, 11:13pm UTC](https://help.galaxyproject.org/t/formatting-annotated-genomes-for-submission-to-ncbi/17716/7 "2026-06-16T23:13:04Z")

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If anyone is interested in the solution to this problem, I found it. The coordinate issue was caused by using NCBI’s FCS-GX, which broke the coordinates relative to the annotations. On Biostars I got a suggestion by Bharat Patel to use a pixi.toml to arrange a series of commands using AGAT and Funannotate to fix the set of problems I described, and the general notion was successful, although I ended up replacing both AGAT and Funannotate with a set of Python scripts. This started with redoing the cleaning step that FCS-GX did – although I was able to use FCS-GX’s action reports alongside the “contamination.txt” from NCBI to guide the cleaning. I could provide more details if anyone is interested.
