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All posts created by jawsWPI

| posted 26 Mar, 2019 15:15
Lokk gene 36 (26423 - 27289 ) | pham 23651
Rachaly_Draft gene 36 (26457 - 27356 ) | pham 23651

Both are reverse genes. Right now there are only 5 members in the pham; two subclusters (A2 and A14); Lokk and CRB1 are the only non-draft genomes.
Edited 26 Mar, 2019 15:24
Posted in: Cluster A Annotation TipsPham 23651 function assignment
| posted 26 Mar, 2019 13:42
What current official function should be assigned to this pham in Rachaly and other drafts? Lokk has called it a “DNA binding domain protein”, CRB1 calls it a putative Rep protein. The research I have done suggests a best fit to “helix-turn-helix DNA binding domain".

I know that Debbie (et.al.?) were going to start investigating this gene and the Par system genes in Lokk and Rachaly back in 2017. Do you have any results that would give us the answer to this question?
Posted in: Cluster A Annotation TipsPham 23651 function assignment
| posted 15 Mar, 2019 14:36
The blast and HHPred evidence suggests that Anakin and NarutoRun's gene 16 (also gene 16 in AnnaSerena and Krampus) is the scaffolding protein–see attached document. AnnaSerena and Krampus assign this as NKF. The synteny is almost correct: it precedes the major capsid, BUT there is an intervening NKF gene; all of these gene are in phams that only belong to EF.

The question is: is this intervening NKF gene enough to disqualify assigning scaffolding protein as the gene 16 function? (I have been told that the scaffolding protein usually immediately precede the major capsid.)
Posted in: Cluster EF Annotation TipsEF Scaffolding protein
| posted 13 Mar, 2019 15:09
After some additional digging there is no substantial evidence for two domains in the alpha chain. So, I'm going to continue the trend and leave these two gene calls with the same function.
Thanks!
Posted in: Cluster EF Annotation TipsTwo piece DnaE-like DNA polymerse III (alpha)
| posted 08 Mar, 2019 22:03
I'm working on two EF draft genomes. All 7 EF genomes have a DnaE-like DNA polymerse III (alpha) "gene" that has been split into two parts. These are genes 56 and 57 in Anakin and NarutoRun, the genes are coding from different frames (+1 and +2), gene 57's RBS Z score is 2.415 it has a so-so final score of -4.264, and there is a consistent 11 bp gap between the genes. The non-draft genomes AnnaSerena and Krampus give both genes the same DnaE-like DNA polymerse III (alpha) function. In an HHPred anaylsis Gene 56 aa1 - 315 has a 100% probability match to aa 19-324 of an Ecoli DnaE DNA polymerase III (alpha) crystal structure, and gene 57 aa15-594 matches (100% probability) the Ecoli structure from 315 - 1160. (If I run HHPred with the merged gene56+57 sequence I again get a 100% probability match but now to the full length Ecoli crystal structure.) I'm seeing the same split, two pieces with the same function whose summed size "equals" an intact DnaE-like DNA polymerse III (alpha), in non-draft (and by phamerator the draft) genomes of several other small clusters: R, AC, CB, DG, and 2-3 singletons. Not all of the pieces are the same size, and in some cases there is a small, intervening "NKF" gene.
The big question is: do I continue the trend and call both functions "DnaE-like DNA polymerse III (alpha)", and if not what is the alternative?
Posted in: Cluster EF Annotation TipsTwo piece DnaE-like DNA polymerse III (alpha)
| posted 27 Nov, 2018 01:06
Is the web site down? A student and I tried opening the site this afternoon–nothing happened. I tried just now (8 pm Monday) and keep getting "502 Bad Gateway".
Thanks. JoAnn
Posted in: Web PhameratorAre we there yet?
| posted 22 Feb, 2018 16:26
Found the issue and solution; still do not know what caused it to happen in the first place.

For some reason DNA master thought there were regions in genes when there were none. It wasn't obvious–when you clicked regions for the affected genes it looked exactly like unaffected genes: a single set of blank boxes for the start, stop and length. However you could click "delete" region in the affected genes and get the correct dialog: i.e. a warning box asking if you wanted to really delete this region, which required a response (yes in my case). I found and deleted all false regions and the validate warning is gone.

SEA experts–any idea why "hidden" single regions might have happened in the first place?
Posted in: DNA MasterValidation error messages
| posted 21 Feb, 2018 19:46
My students are seeing a very weird validation result. It wasn't present in the original merged file which showed fixable or expected messages, but appeared later after students started editing the merged files (attachment). I have tried everything I could think of to get rid of this: re-numbered, re-tagged, recreate documentation, saved and reopen (separately in sequence) and still have the weird
"join(0. .0)
Region lengths not correct
complement (join(0. .0))
Region lengths not correct" messages.

Has anyone else seen this? Is it an issue for final QC? Is there a solution out there?
Posted in: DNA MasterValidation error messages
| posted 21 Feb, 2018 19:21
Welkin Pope
Hi Joann,
Did you remember to change the number of regions in the description tab to "2" ?

Best,
Welkin

I just double checked–and the number of regions in the description tab is set to 2. Still see same validation error message(s). We are also seeing "15102 - 15473 and share a 5' (upstream) coordinate"–am assuming is a valid warning since the properly annotated longer (downstream) gene will share a start site with the upstream gene.

I've tried several other things to try and get rid of the following without success.
"join(0. .0;0. .0)
Incorrect region counts"
or "complement (join(0. .0))
Region lengths not correct"

Looks like this is a different issue. I'll create a new topic (validation errors) and will post more details there.
Posted in: Frameshifts and IntronsAnnotation advise: Frameshift in A4
| posted 20 Feb, 2018 21:52
This is what the annotation looks like.
Posted in: Frameshifts and IntronsAnnotation advise: Frameshift in A4