[maker-devel] substr outside of string in PhatHits_utils.pm

[Ole Kristian Tørresen]

Thank you Carson.

After a bit of struggling, I can confirm that the same error occurs in MAKER 3.01.2 (I guess you meant that version, couldn’t find 3.02.02).

I am providing a GFF to est_gff, with match and match_part entries. For at least one of the scaffolds, the last coordinate (column 5) is the same number as the length of the scaffold. That should be allowed by the GFF3 standard, right?

How can I troubleshoot this? The error message is not so informative. It seems that PhatHit_utils.pm tries to find a stop codon. Snipped from that file, lines 849-850:
#fix stop codon by walking downstream
my $has_stop = $tM->is_ter_codon(substr($transcript_seq, $end-1-3, 3));

The GFF I am using was the output of Mikado (https://www.biorxiv.org/content/early/2017/11/09/216994), which is GFF3, and then processed a bit to make it suitable for MAKER. First converted to GTF by 'mikado util convert mikado.loci.gff3 mikado.loci.gtf'

Then I selected only mRNA and exon entries, and changed mRNA to transcript to make it look like cufflinks output (and set a dummy score):
grep -P "\tmRNA\t|\texon\t" mikado.loci.gtf |sed "s/mRNA/transcript/g" |awk -F "\t" '{$9=$9"cov \"10.0\";"; OFS="\t"; print $1, $2, $3, $4, $5, $6, $7, $8, $9}' > mikado.loci.score.gtf

Before converting with cufflinks2gff3:
cufflinks2gff3 mikado.loci.score.gtf > ests.score.gff3

Thank you.

Ole


> On 09 Nov 2017, at 17:28, Carson Holt <carsonhh at gmail.com> wrote:
> 
> My first guess is that if you are using gff3 files as input to anything, then there may be an  issue with your GFF3 file. My second suggestion is to try MAKER 3.02.02 to see if it has the same issue.
> 
> —Carson
> 
> 
>> On Nov 9, 2017, at 2:44 AM, Ole Kristian Tørresen <o.k.torresen at ibv.uio.no> wrote:
>> 
>> Dear all,
>> I'm having an issue with MAKER which I'm unable to wrap my head around. Hopefully the issue is easily identifiable and resolvable for someone with more insight than me. Please find the log output attached below. I cannot find any more information than this in any logs. Many scaffolds do complete fine, but some of the longest ones have issues.
>> 
>> Thank you.
>> 
>> Sincerely,
>> Ole K. Tørresen
>> 
>> Error message:
>> 
>> #--------- command -------------#
>> Widget::augustus:
>> /projects/cees/bin/augustus/augustus-3.2.3/bin/augustus  --strand=backward --species=gadMor2_code_braker2 --UTR=off --hintsfile=/tmp/18899594.d/maker_m8xwVp/0/34_2.22239-28465.gadMor2_code_brak
>> er2.auto_annotator.xdef.augustus --extrinsicCfgFile=/projects/cees/bin/augustus/augustus-3.2.3/config/extrinsic/extrinsic.MPE.cfg --AUGUSTUS_CONFIG_PATH=/projects/cees/bin/augustus/augustus-3.2
>> .3/config /tmp/18899594.d/maker_m8xwVp/0/34_2.22239-28465.gadMor2_code_braker2.auto_annotator.augustus.fasta > /tmp/18899594.d/maker_m8xwVp/0/34_2.22239-28465.gadMor2_code_braker2.auto_annotato
>> r.augustus
>> #-------------------------------#
>> deleted:0 genes
>> begin called get_best_alt_splices1
>> ...processing 0 of 2
>> ...processing 1 of 2
>> end called get_best_alt_splices1
>> ...processing 0 of 20
>> ...processing 1 of 20
>> ...processing 2 of 20
>> ...processing 3 of 20
>> ...processing 4 of 20
>> ...processing 5 of 20
>> ...processing 6 of 20
>> ...processing 7 of 20
>> ...processing 8 of 20
>> ...processing 9 of 20
>> ...processing 10 of 20
>> ...processing 11 of 20
>> ...processing 12 of 20
>> ...processing 13 of 20
>> ...processing 14 of 20
>> ...processing 15 of 20
>> ...processing 16 of 20
>> ...processing 17 of 20
>> ...processing 18 of 20
>> ...processing 19 of 20
>> substr outside of string at /projects/cees/bin/maker/maker-3.1.1/bin/../lib/PhatHit_utils.pm line 850.
>> --> rank=NA, hostname=compute-31-18.local
>> ERROR: Failed while annotating transcripts
>> ERROR: Chunk failed at level:1, tier_type:4
>> FAILED CONTIG:GmG20150304_scaffold_8692
>> 
>> ERROR: Chunk failed at level:6, tier_type:0
>> FAILED CONTIG:GmG20150304_scaffold_8692
>> 
>> examining contents of the fasta file and run log
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>