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All posts created by kcornely

| posted 11 Sep, 2025 11:37
cdshaffer
just to document for anyone what wants to create an auto-annotation here is what I did:

1. open DNA Master and do the standard File -> Open Fasta and import the sequence
2. Go go https://discover.kbrinsgd.org/autoannotate/ and enter phage name and upload the same sequence file you used in step 1
3. Click "Process"
4. In the text box below the process button you will get the auto-annotation as "Documentation"; it will have a bunch of entries that look something like this:

CDS 15 - 272
  /gene="1"
  /product="gp1"
  /locus tag="Roots515_1"

5. Select and Copy the entire contents of that text box
6. Go back to DNA master, on the phage window click the "Documentation" tab at the top right
7. Delete any text in the large text box and paste in from the results from the PECAAN auto-annotation. (When I did this this step I got about 100 question marks at the end which I deleted so the last entry ended like this:

CDS join(155929. .156288;1. .1smile
  /gene="232"
  /product="gp232"
  /locus tag="Roots515_232"
    /note=Original Glimmer call @bp 155929 has strength 11.01 ** not called by GeneMark

8. click the parse button, then in the window that opens accept the defaults and click the new parse button. This will convert the "documentation" into DNA Master gene entries
9. Save the DNA Master file as usual

Greetings, all!
I posted a comment last night, but it appears that I didn't quote the correct post, so I am replying again. We were having trouble with the auto annotation. We got a "Glimmer error" message, but the result was that we got all of the Glimmer calls, but every feature said "not called by Glimmer". We used PECAAN to do the auto annotation, and then we used Chris's directions here to re-create the documentation in DNA Master, and it worked! We now have both calls for all features. I then BLASTed the entire genome overnight and everything look great. It's nice that these old posts are still on the forums and that they still work!
Thanks,
Kathleen
Posted in: DNA MasterGlimmer Failure on Auto Annotation
| posted 11 Sep, 2025 03:28
cdshaffer
I am curious, have you tried manually testing the NCBI glimmer page to see if it is working when you are getting the failures when trying to access with DNA Master?

https://www.ncbi.nlm.nih.gov/genomes/MICROBES/glimmer_3.cgi
If that page is down then it is guaranteed DNA Master will fail.

Also, have you tried increasing the time limit in the auto-annotation window? It defaults to 2 minutes, maybe if NCBI is slowing down responses setting DNA Master to wait longer would help.

Greetings, all!
I know that this is an old post, but we were having trouble with the auto annotation. During auto annotation, Glimmer calls were made, but every single feature listed "not called by GeneMark". We checked, double-checked and triple-checked our preferences, and that didn't work. I used the directions posted here and it was a great work-around. Thanks, Chris!
Posted in: DNA MasterGlimmer Failure on Auto Annotation
| posted 04 Feb, 2025 22:21
Thanks, Vic, this is very helpful!
Posted in: MycobacteriumSource of albumin for AD
| posted 04 Feb, 2025 15:55
Greetings, Mycobacterium smegmatis community! I normally purchase my albumin to make AD from Spectrum. I've never had a problem with them in the past, but I placed an order with them in November that is backordered until the end of February. The back order date keeps changing, so I finally asked them if they REALLY knew that the albumin would be shipped at the end of February, or if they were just being hopeful and they were making up these dates. Turns out they are making up the backorder dates and they have no idea when the albumin will arrive. I have enough to last me at least a month, but could someone recommend another supplier for me? Thanks!
Posted in: MycobacteriumSource of albumin for AD
| posted 30 Jul, 2023 22:55
Is it possible for a phage to have two WhiB family transcription factors? We think we've identified two in phage Maravista.
Posted in: Functional AnnotationWhiB family transcription factors
| posted 30 Jul, 2023 22:09
Greetings, all! We are getting ready to submit our annotation for Maravista and we haven't received a reply to this query, so I hope someone can help us out. To clarify, it's a reverse gene and it's gp 41 on phagesdb and we are going with the GeneMark start at 32125. We're fairly certain it's a transposase but want to be sure. Thanks!
Posted in: Functional AnnotationIdentify transposons
| posted 23 Jun, 2023 18:47
Greetings, all,
Is anyone else having trouble updating DNA Master? We get an error message when we attempt to update. I've attached a screenshot. We've tried computers on campus and off campus, laptops and desk tops, and nothing is working. The update reaches 33%, and then DNA Master either freezes, or we get an FTP error. It appears that DNA Master is having difficulty connecting to the server. Any advice to solve this problem would be appreciated and thanks in advance!
Posted in: Bioinformatic Tools and AnalysesDNA Master not updating
| posted 22 Jun, 2023 16:47
We are annotating a cluster F1 phage (Maravista)and have identified a transposase (gp 41) and we need to find the corresponding transposon. We haven't done this before. What tools would be effective to assist us in identifying a transposon?
Thanks in advance for the help!
Posted in: Functional AnnotationIdentify transposons
| posted 09 Jun, 2023 18:15
Talking to Debbie in person! We clarified that the start site is the second methionine in the sequence, which means that the translated protein has a single initial methionine and not two Mets, irrespective of the direction of translation.
Edited 09 Jun, 2023 19:25
Posted in: Cluster P Annotation TipsTwo methionines at the N-terminus?
| posted 02 May, 2023 02:26
Greetings, all! We are annotating a cluster A3 mycobacteriophage and we see that the lysin A and lysin B proteins do not follow the expected synteny in which lysin A, then holin, and then lysin B follow the minor tail proteins expressed downstream of the tape measure protein. Instead we see lysin A and lysin B near the 5' end of the genome, expressed upstream of terminase. And we aren't able to find a protein we can assign as a holin. Is synteny different in cluster A3 phages? Thanks in advance for your help!
Posted in: Cluster A Annotation TipsLysins A and B not following synteny