<div dir="ltr"><div style="font-family:arial,sans-serif;font-size:13px">Dear Ensembl team,</div><div style="font-family:arial,sans-serif;font-size:13px"><br></div><div style="font-family:arial,sans-serif;font-size:13px">I am currently working with a GFF3 annotation of Triticum aestivum (MIPS v2.1, Ensembl v22).</div>
<div style="font-family:arial,sans-serif;font-size:13px"> <span dir="ltr"><a href="ftp://ftp.ensemblgenomes.org/pub/plants/release-22/gff3/triticum_aestivum/Triticum_aestivum.IWGSP1.22.gff3.gz" target="_blank">ftp://ftp.ensemblgenomes.org/pub/plants/release-22/gff3/triticum_aestivum/Triticum_aestivum.IWGSP1.22.gff3.gz</a></span><br>
</div><div style="font-family:arial,sans-serif;font-size:13px"><br></div><span style="font-family:arial,sans-serif;font-size:13px">In our study, we are examining single nucleotide polymorphism variants in wheat and their effect on protein coding regions using SNPeff 3.5d (build 2014-03-05)</span><span style="font-family:arial,sans-serif;font-size:13px"><div>
<br></div><div>I see a warning message from SNPeff "WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS" in 17,834 genes. Below are 10 examples of genes that have this warning and 10 that do not.</div><div><br></div><div>Do you know what might be causing such warning? Or refer me to someone who can help?</div>
<div><br></div><div>Thank you in advance.</div><div><br></div><div>Best wishes,</div><div><br></div><div>Ksenia</div><div><br></div></span><span style="font-family:arial,sans-serif;font-size:13px"><div><span style="font-family:arial;font-size:small">-- </span><br style="font-family:arial;font-size:small">
<div dir="ltr" style="font-family:arial;font-size:small">Ksenia Krasileva, PhD<br><br>Post Doctoral Scholar<br>USDA NIFA Fellowship<br>The Dubcovsky Lab<br><br>Department of Plant Sciences<br>University of California, Davis<br>
124 Robbins Hall<br>Davis, CA 95616<br><a href="mailto:krasileva@ucdavis.edu" target="_blank">krasileva@ucdavis.edu</a></div></div><div dir="ltr" style="font-family:arial;font-size:small"><br></div><div style="font-family:arial;font-size:small">
SNPeff data:</div><div style="font-family:arial;font-size:small"><br></div>10 examples without warnings:</span><br style="font-family:arial,sans-serif;font-size:13px"><div style="font-family:arial,sans-serif;font-size:13px">
IWGSC_CSS_5BS_scaff_1487662     672     Kronos381       G       A       40.0    Pass    seed_avail=Kronos381;EFF=SYNONYMOUS_CODING(LOW|SILENT|ggC/ggT|G280||Traes_5BS_38DA1CBC61.E1|||transcript:Traes_5BS_38DA1CBC61.2|1|1)</div>
<div style="font-family:arial,sans-serif;font-size:13px">IWGSC_CSS_5BS_scaff_1537048     1688    Kronos244       C       T       40.0    Pass    seed_avail=Kronos244;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|gCt/gTt|A58V||CDS:Traes_5BS_B2CD001D1.2|||transcript:Traes_5BS_B2CD001D1.2|2|1)</div>
<div style="font-family:arial,sans-serif;font-size:13px">IWGSC_CSS_5BS_scaff_1537048     1718    Kronos439       C       T       40.0    Pass    seed_avail=Kronos439;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|gCt/gTt|A68V||CDS:Traes_5BS_B2CD001D1.2|||transcript:Traes_5BS_B2CD001D1.2|2|1)</div>
<div style="font-family:arial,sans-serif;font-size:13px">IWGSC_CSS_5BS_scaff_1646558     3136    Kronos684       G       A       40.0    Pass    seed_avail=Kronos684;EFF=SYNONYMOUS_CODING(LOW|SILENT|acC/acT|T115||Traes_5BS_E2116F66A.E1|||transcript:Traes_5BS_E2116F66A.1|1|1)</div>
<div style="font-family:arial,sans-serif;font-size:13px">IWGSC_CSS_5BS_scaff_168385      2625    Kronos2646      G       A       40.0    Pass    seed_avail=Kronos2646;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|Gag/Aag|E18K||Traes_5BS_91BD3E004.E1|||transcript:Traes_5BS_91BD3E004.1|1|1)</div>
<div style="font-family:arial,sans-serif;font-size:13px">IWGSC_CSS_5BS_scaff_168385      2666    Kronos390       C       T       40.0    Pass    seed_avail=Kronos390;EFF=SYNONYMOUS_CODING(LOW|SILENT|gcC/gcT|A31||Traes_5BS_91BD3E004.E1|||transcript:Traes_5BS_91BD3E004.1|1|1)</div>
<div style="font-family:arial,sans-serif;font-size:13px">IWGSC_CSS_5BS_scaff_168385      2714    Kronos166       G       A       40.0    Pass    seed_avail=Kronos166;EFF=SYNONYMOUS_CODING(LOW|SILENT|gaG/gaA|E47||Traes_5BS_91BD3E004.E1|||transcript:Traes_5BS_91BD3E004.1|1|1)</div>
<div style="font-family:arial,sans-serif;font-size:13px">IWGSC_CSS_5BS_scaff_1712494     1012    Kronos447       C       T       40.0    Pass    seed_avail=Kronos447;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|Ggg/Agg|G681R||Traes_5BS_6004CA039.E3|||transcript:Traes_5BS_6004CA039.1|1|1)</div>
<div style="font-family:arial,sans-serif;font-size:13px">IWGSC_CSS_5BS_scaff_1712494     2037    Kronos2214      G       A       40.0    Pass    seed_avail=Kronos2214;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|tCc/tTc|S339F||Traes_5BS_6004CA039.E3|||transcript:Traes_5BS_6004CA039.1|1|1)</div>
<div style="font-family:arial,sans-serif;font-size:13px">IWGSC_CSS_5BS_scaff_1712494     2086    Kronos199       G       A       40.0    Pass    seed_avail=Kronos199;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|Cct/Tct|P323S||Traes_5BS_6004CA039.E3|||transcript:Traes_5BS_6004CA039.1|1|1)</div>
<div style="font-family:arial,sans-serif;font-size:13px"><br></div><div style="font-family:arial,sans-serif;font-size:13px"><br></div><div style="font-family:arial,sans-serif;font-size:13px">10 Examples with warnings:<br>
<div><div>IWGSC_CSS_1BS_scaff_1134305     892     Kronos2092      G       A       40.0    Pass    seed_avail=Kronos2092;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|Gtt/Att|V123I||Traes_1BS_9AD3B3BE2.E22|||transcript:Traes_1BS_9AD3B3BE2.1|4|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS),SPLICE_SITE_REGION(LOW|||||Traes_1BS_9AD3B3BE2.E22|||transcript:Traes_1BS_9AD3B3BE2.1|4|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS)</div>
<div>IWGSC_CSS_1BS_scaff_1134305     5997    Kronos2711      C       T       40.0    Pass    seed_avail=Kronos2711;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|gCt/gTt|A806V||Traes_1BS_9AD3B3BE2.E22|||transcript:Traes_1BS_9AD3B3BE2.1|18|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS)</div>
<div>IWGSC_CSS_1BS_scaff_1260855     1972    Kronos375       G       A       40.0    Pass    seed_avail=Kronos375;EFF=SYNONYMOUS_CODING(LOW|SILENT|cgG/cgA|R266||Traes_1BS_91DEF138E.E1|||transcript:Traes_1BS_91DEF138E.1|6|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS)</div>
<div>IWGSC_CSS_1BS_scaff_1280056     767     Kronos166       C       T       40.0    Pass    seed_avail=Kronos166;EFF=SYNONYMOUS_CODING(LOW|SILENT|aaC/aaT|N93||Traes_1BS_984B78A41.E3|||transcript:Traes_1BS_984B78A41.1|3|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS)</div>
<div>IWGSC_CSS_1BS_scaff_1420918     702     Kronos209       G       A       40.0    Pass    seed_avail=Kronos209;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|gGg/gAg|G138E||Traes_1BS_4073CE3DB.E34|||transcript:Traes_1BS_4073CE3DB.1|1|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS)</div>
<div>IWGSC_CSS_1BS_scaff_1420918     871     Kronos166       G       A       40.0    Pass    seed_avail=Kronos166;EFF=SYNONYMOUS_CODING(LOW|SILENT|agG/agA|R194||Traes_1BS_4073CE3DB.E34|||transcript:Traes_1BS_4073CE3DB.1|1|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS)</div>
<div>IWGSC_CSS_1BS_scaff_1420918     912     Kronos209       G       A       40.0    Pass    seed_avail=Kronos209;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|gGa/gAa|G208E||Traes_1BS_4073CE3DB.E34|||transcript:Traes_1BS_4073CE3DB.1|1|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS)</div>
<div>IWGSC_CSS_1BS_scaff_1420918     5050    Kronos2553      G       A       40.0    Pass    seed_avail=Kronos2553;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|Gta/Ata|V692I||Traes_1BS_4073CE3DB.E34|||transcript:Traes_1BS_4073CE3DB.1|11|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS),UPSTREAM(MODIFIER||1659|||Traes_1BS_587F929ED.E1|||transcript:Traes_1BS_587F929ED.1||1)</div>
<div>IWGSC_CSS_1BS_scaff_1912213     607     Kronos910       G       A       40.0    Pass    seed_avail=Kronos910;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|aGt/aAt|S59N||Traes_1BS_0039045F7.E21|||transcript:Traes_1BS_0039045F7.2|1|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS)</div>
<div>IWGSC_CSS_1BS_scaff_1975885     313     Kronos684       G       A       40.0    Pass    seed_avail=Kronos684;EFF=NON_SYNONYMOUS_CODING(MODERATE|MISSENSE|Gga/Aga|G25R||Traes_1BS_CC1B13C13.E32|||transcript:Traes_1BS_CC1B13C13.1|1|1|WARNING_TRANSCRIPT_MULTIPLE_STOP_CODONS)</div>
</div></div><div><br></div>-- <br><div dir="ltr">Ksenia Krasileva, PhD<br><br>Post Doctoral Scholar<br>USDA NIFA Fellowship<br>The Dubcovsky Lab<br><br>Department of Plant Sciences<br>University of California, Davis<br>124 Robbins Hall<br>
Davis, CA 95616<br><a href="mailto:krasileva@ucdavis.edu" target="_blank">krasileva@ucdavis.edu</a><br></div>
</div>