IKMC Project Report - EUCOMM (ID: 28317)

2310067B10Rik MGI:1919197 ENSMUSG00000020747 OTTMUSG00000003538
Pre-pipeline Designs Vectors ES Cells Mice
Mice - Genotype confirmed

Mice

Microinjection Status Allele Allele Type ES Cell Clone Genetic Background QC Data MI/Distribution Centre
contact distributor Genotype confirmed 2310067B10Riktm1a(EUCOMM)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) EPD0215_1_D09 C57BL/6NTac;C57BL/6NTac;C57BL/6N view
( about ) 		ICS
Southern Blot na TV Backbone Assay na 5' LR-PCR na
Loss of WT Allele (LOA) qPCR na Homozygous Loss of WT Allele (LOA) SR-PCR na Neo Count (qPCR) na
LacZ SR-PCR pass 5' Cassette Integrity Neo SR-PCR na
Mutant Specific SR-PCR pass LoxP Confirmation pass 3' LR-PCR
Genotyping Comment -

Mutagenesis Predictions

This gene has 7 wild type transcripts, of which 6 are protein-coding. Following removal of the floxed region, 6 transcripts are predicted to produce a truncated protein product of which 2 may be subject to non-sense mediated decay (NMD). The original allele for this mutation is of type 'Knockout First, Reporter-tagged insertion with conditional potential'. The table below shows the predicted structure of the gene transcripts after application of Flp and Cre (forming a 'Knockout-First, Post-Flp and Cre - Deletion, No Reporter' allele - more information on IKMC alleles can be found here). Click the 'view' button for each transcript to see the full prediction for that transcript.

Ensembl transcript id Ensembl Biotype Floxed transcript description Details
ENSMUST00000093912 protein_coding No protein product (NMD) view

Predicted translation: 

MDLREKHLGEPPLALGLSTRKALSVLKEQLEAVLEKHLKERKKSLTWKEAWRSSFLHLSNRCSCFHWPGASLMLLAVLLL
LCCCGGQPAGSQGVELVNASALFLLLLLNLVLIGRQDRLKRREVERRLRGIIDQIQGR*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000661085 UTR UTR
ENSMUSE00000585515 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00000585514 CDS CDS
ENSMUSE00001029606 CDS CDS
ENSMUSE00000585512 CDS CDS
ENSMUSE00000585511 CDS Deleted
ENSMUSE00000585510 CDS CDS + stop codon + UTR
ENSMUSE00000585509 CDS
ENSMUSE00000661083 CDS
ENSMUSE00000585508 CDS
ENSMUSE00000585507 CDS
ENSMUSE00000585506 CDS
ENSMUSE00000585505 CDS
ENSMUSE00000585504 CDS
ENSMUSE00000585503 CDS
ENSMUSE00000585502 CDS
ENSMUSE00000585501 CDS
ENSMUSE00000585500 CDS
ENSMUSE00000585499 CDS
ENSMUSE00000585498 CDS
ENSMUSE00000585497 CDS
ENSMUSE00000585496 CDS
ENSMUSE00000585495 CDS
ENSMUSE00000585494 CDS
ENSMUSE00000585493 CDS
ENSMUSE00000585492 ATPase_P-typ_cation-transptr_C (19/213 aa) CDS
ENSMUSE00000585491 ATPase_P-typ_cation-transptr_C (28/213 aa) CDS
ENSMUSE00000585490 ATPase_P-typ_cation-transptr_C (50/213 aa) CDS
ENSMUSE00000585489 ATPase_P-typ_cation-transptr_C (33/213 aa) CDS
ENSMUSE00000585488 ATPase_P-typ_cation-transptr_C (32/213 aa) CDS
ENSMUSE00000585487 ATPase_P-typ_cation-transptr_C (55/213 aa) CDS
ENSMUSE00000477452 CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000103033 protein_coding No protein product (NMD) view

Predicted translation: 

MDLREKHLGEPPLALGLSTRKALSVLKEQLEAVLEKHLKERKKSLTWKEAWRSSFLHLSNRCSCFHWPGASLMLLAVLLL
LCCCGGQPAGSQGVELVNASALFLLLLLNLVLIGRQDRLKRREVERRLRGIIDQIQGR*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000463190 UTR UTR
ENSMUSE00000471761 UTR UTR
ENSMUSE00000438163 UTR UTR
ENSMUSE00000585515 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00000585514 CDS CDS
ENSMUSE00001029606 CDS CDS
ENSMUSE00000585512 CDS CDS
ENSMUSE00000585511 CDS Deleted
ENSMUSE00000585510 CDS CDS + stop codon + UTR
ENSMUSE00000585509 CDS
ENSMUSE00000661083 CDS
ENSMUSE00000585508 CDS
ENSMUSE00000585507 CDS
ENSMUSE00000585506 CDS
ENSMUSE00000585505 CDS
ENSMUSE00000585504 CDS
ENSMUSE00000585503 CDS
ENSMUSE00000585502 CDS
ENSMUSE00000585501 CDS
ENSMUSE00000585500 CDS
ENSMUSE00000585499 CDS
ENSMUSE00000585498 CDS
ENSMUSE00000585497 CDS
ENSMUSE00000585496 CDS
ENSMUSE00000585495 CDS
ENSMUSE00000585494 CDS
ENSMUSE00000585493 CDS
ENSMUSE00000585492 ATPase_P-typ_cation-transptr_C (19/213 aa) CDS
ENSMUSE00000585491 ATPase_P-typ_cation-transptr_C (28/213 aa) CDS
ENSMUSE00000585490 ATPase_P-typ_cation-transptr_C (50/213 aa) CDS
ENSMUSE00000585489 ATPase_P-typ_cation-transptr_C (33/213 aa) CDS
ENSMUSE00000585488 ATPase_P-typ_cation-transptr_C (32/213 aa) CDS
ENSMUSE00000585487 ATPase_P-typ_cation-transptr_C (55/213 aa) CDS
ENSMUSE00000585486 CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000141871 protein_coding Target region does not overlap transcript
ENSMUST00000136720 protein_coding Target region does not overlap transcript
ENSMUST00000103034 protein_coding Target region does not overlap transcript
ENSMUST00000125918 protein_coding Target region does not overlap transcript
ENSMUST00000134948 processed_transcript Target region does not overlap transcript
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ES Cell Clones With Knockout First Mutation (Reporter Tagged Insertion With Conditional Potential)

Genome Browsers: Ensembl (mouse) - UCSC (mouse)
Tools: Southern Blot Tool - LRPCR Genotyping Primers
ES Cell Clone Targeting Vector Allele Allele Type Parental ES Cell Line Genbank File Mouse QC Data
order EPD0215_1_D09 PGRS0004_A_B02 2310067B10Riktm1a(EUCOMM)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8.N4 view yes view    ( about )
Production Centre
5' Screen not attempted LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
LOA - LOXP - LACZ -
Chromosome 1 - Chromosome 8a - Chromosome 8b -
Chromosome 11a - Chromosome 11b - Chromosome Y -
User/Mouse Clinic
Southern Blot pass Map Test - Karyotype -
TV Backbone Assay - 5' LR-PCR - Loss of WT Allele (LOA) -
Neo Count (qPCR) - LacZ SR-PCR - 5' Cassette Integrity -
Neo SR-PCR - Mutant Specific SR-PCR - LoxP Confirmation -
3' LR-PCR -
order EPD0215_1_A10 PGRS0004_A_B02 2310067B10Riktm1a(EUCOMM)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8.N4 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
LOA - LOXP - LACZ -
Chromosome 1 - Chromosome 8a - Chromosome 8b -
Chromosome 11a - Chromosome 11b - Chromosome Y -
User/Mouse Clinic
Southern Blot - Map Test - Karyotype -
TV Backbone Assay - 5' LR-PCR - Loss of WT Allele (LOA) -
Neo Count (qPCR) - LacZ SR-PCR - 5' Cassette Integrity -
Neo SR-PCR - Mutant Specific SR-PCR - LoxP Confirmation -
3' LR-PCR -
order EPD0215_1_D10 PGRS0004_A_B02 2310067B10Riktm1a(EUCOMM)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8.N4 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
LOA - LOXP - LACZ -
Chromosome 1 - Chromosome 8a - Chromosome 8b -
Chromosome 11a - Chromosome 11b - Chromosome Y -
User/Mouse Clinic
Southern Blot - Map Test - Karyotype -
TV Backbone Assay - 5' LR-PCR - Loss of WT Allele (LOA) -
Neo Count (qPCR) - LacZ SR-PCR - 5' Cassette Integrity -
Neo SR-PCR - Mutant Specific SR-PCR - LoxP Confirmation -
3' LR-PCR -
order EPD0215_1_D11 PGRS0004_A_B02 2310067B10Riktm1a(EUCOMM)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8.N4 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
LOA - LOXP - LACZ -
Chromosome 1 - Chromosome 8a - Chromosome 8b -
Chromosome 11a - Chromosome 11b - Chromosome Y -
User/Mouse Clinic
Southern Blot - Map Test - Karyotype -
TV Backbone Assay - 5' LR-PCR - Loss of WT Allele (LOA) -
Neo Count (qPCR) - LacZ SR-PCR - 5' Cassette Integrity -
Neo SR-PCR - Mutant Specific SR-PCR - LoxP Confirmation -
3' LR-PCR -
order EPD0215_1_H12 PGRS0004_A_B02 2310067B10Riktm1a(EUCOMM)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) JM8.N4 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen pass 3' Screen pass
Loss of WT Allele (LOA) - Vector Integrity -
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
LOA - LOXP - LACZ -
Chromosome 1 - Chromosome 8a - Chromosome 8b -
Chromosome 11a - Chromosome 11b - Chromosome Y -
User/Mouse Clinic
Southern Blot - Map Test - Karyotype -
TV Backbone Assay - 5' LR-PCR - Loss of WT Allele (LOA) -
Neo Count (qPCR) - LacZ SR-PCR - 5' Cassette Integrity -
Neo SR-PCR - Mutant Specific SR-PCR - LoxP Confirmation -
3' LR-PCR -
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Targeting Vectors

Genome Browsers: Ensembl (mouse) - UCSC (mouse)
Tools: LRPCR Genotyping Primers
Design ID Vector Type Targeting Vector Cassette Backbone Genbank File
order 141 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PGR00004_A_2_B02 L1L2_Bact_P L4L3_pD223_DTA_spec view
order 141 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PGR00004_A_3_B02 L1L2_Bact_P L4L3_pD223_DTA_spec view
order 141 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PGR00004_A_4_B02 L1L2_Bact_P L4L3_pD223_DTA_spec view
order 141 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PGRS0004_A_B02 L1L2_Bact_P L4L3_pD223_DTA_spec view
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Intermediate Vectors

Design ID Vector Type Intermediate Vector
141 Knockout First, Reporter-tagged insertion with conditional potential PCS00035_A_C10