IKMC Project Report - KOMP-CSD (ID: 37405)

1700067K01Rik MGI:1920703 ENSMUSG00000046408 OTTMUSG00000028911
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 1700067K01Riktm2a(KOMP)Wtsi Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) EPD0180_3_C06 C57BL/6NTac;C57BL/6NTac;C57BL/6N view
( about ) 		WTSI
Southern Blot na TV Backbone Assay pass 5' LR-PCR na
Loss of WT Allele (LOA) qPCR pass Homozygous Loss of WT Allele (LOA) SR-PCR na Neo Count (qPCR) pass
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 5 wild type transcripts, of which 3 are protein-coding. Following removal of the floxed region, 3 transcripts are predicted to produce a truncated protein product of which 0 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
ENSMUST00000060357 protein_coding Residual N-terminal, residual C-terminal product view

Predicted translation: 

MMEAPASTSSREPLAAELGPAPARLPLDSMFSPITDQLRYLLRKADDFQSYLLYSYFLCYRDTWEDAGQGPANSCPQIQK
LWSIGRWMPLGPAEDDLDSWILCPQPPGDYQQLLTIGFEEPSHVLATDLLVQILLGQAGPARPPSAAGPAAWAAPAS*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000344319 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00000343781 CDS Deleted
ENSMUSE00000389527 DUF3314 (16/158 aa) CDS Deleted
ENSMUSE00000407906 DUF3314 (63/158 aa) CDS Deleted
ENSMUSE00000369798 DUF3314 (46/158 aa) CDS CDS
ENSMUSE00000410423 DUF3314 (36/158 aa) CDS + stop codon + UTR CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000172548 protein_coding Residual N-terminal, residual C-terminal product view

Predicted translation: 

MMEAPASTSSREPLAAELGPAPARLPLDSMFSPITDQLRYLLRKADDFQSYLLYSYFLCYRDTWEDAGQGPANSCPQIQK
LWSIGRWMPLGPAEDDLDSWILCPQPPGDYQQLLTIGFEEPSHVLATDLLVQILLGQAGPARPPSAAGPAAWAAPAS*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000943829 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00000943438 CDS Deleted
ENSMUSE00000389527 DUF3314 (16/158 aa) CDS Deleted
ENSMUSE00000407906 DUF3314 (63/158 aa) CDS Deleted
ENSMUSE00000369798 DUF3314 (46/158 aa) CDS CDS
ENSMUSE00000928036 DUF3314 (36/158 aa) CDS + stop codon + UTR CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000174587 protein_coding Residual N-terminal, residual C-terminal product view

Predicted translation: 

MFSPITDQLRYLLRKADDFQSYLLYSYFLCYRDTWEDAGQGPANSCPQIQKLWSIGRWMPLGPAEDDLDSWILCPQPPGD
YQQLLTIGFEEPSHVLATDLLVQILLGQAGPARPPSAAGPAAWAAPAS*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000930728 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00000343781 CDS Deleted
ENSMUSE00000389527 DUF3314 (16/158 aa) CDS Deleted
ENSMUSE00000407906 DUF3314 (63/158 aa) CDS Deleted
ENSMUSE00000369798 DUF3314 (46/158 aa) CDS CDS
ENSMUSE00000410423 DUF3314 (36/158 aa) CDS + stop codon + UTR CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000174570 nonsense_mediated_decay No protein product (NMD) view

Predicted translation: 

MMEAPASTSSREPLAAELGPAPARLPLDSMFSPITDQLRYLLRKADDFQSYLLYSYFLCYRDTWEDAGQGPANSCPQIQK
LWSIGRWMPLGPAEDDLDSW*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00000943829 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00000943438 CDS Deleted
ENSMUSE00000389527 DUF3314 (16/124 aa) CDS Deleted
ENSMUSE00000407906 DUF3314 (63/124 aa) CDS Deleted
ENSMUSE00000928981 DUF3314 (46/124 aa) CDS + stop codon + UTR CDS + stop codon + UTR
ENSMUSE00000929869 UTR UTR
Legend: in frame deleted frameshifted
ENSMUST00000140120 retained_intron No analysis available: incomplete 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 EPD0180_3_C06 PRPGS00066_B_G07 1700067K01Riktm2a(KOMP)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 not confirmed
Loss of WT Allele (LOA) pass Vector Integrity -
Distribution Centre
Karyotype - Copy Number - Cells Thawed Correctly -
5' LR-PCR - 5' SR-PCR - 3' SR-PCR -
LOA pass LOXP pass LACZ pass
Chromosome 1 pass Chromosome 8a pass Chromosome 8b -
Chromosome 11a - Chromosome 11b pass Chromosome Y pass
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 EPD0180_3_A05 PRPGS00066_B_G07 1700067K01Riktm2a(KOMP)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 not confirmed
Loss of WT Allele (LOA) pass 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 EPD0180_3_B07 PRPGS00066_B_G07 1700067K01Riktm2a(KOMP)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 not confirmed
Loss of WT Allele (LOA) pass 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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ES Cell Clones Without Conditional Potential

Note: Mutations of type "Without Conditional Potential" are correctly targeted clones that have lost the 3' LoxP site. These mutations cannot be converted into conditional alleles.

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 EPD0180_3_C05 PRPGS00066_B_G07 1700067K01Riktm2e(KOMP)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8.N4 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen no reads detected 3' Screen no reads detected
Loss of WT Allele (LOA) pass 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 EPD0180_3_D05 PRPGS00066_B_G07 1700067K01Riktm2e(KOMP)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8.N4 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen not confirmed 3' Screen not confirmed
Loss of WT Allele (LOA) pass 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 EPD0180_3_D08 PRPGS00066_B_G07 1700067K01Riktm2e(KOMP)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8.N4 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen not confirmed 3' Screen not confirmed
Loss of WT Allele (LOA) pass 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 EPD0180_3_G07 PRPGS00066_B_G07 1700067K01Riktm2e(KOMP)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8.N4 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen no reads detected 3' Screen no reads detected
Loss of WT Allele (LOA) pass 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 EPD0180_3_H07 PRPGS00066_B_G07 1700067K01Riktm2e(KOMP)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8.N4 view no view    ( about )
Production Centre
5' Screen not attempted LoxP Screen not confirmed 3' Screen not confirmed
Loss of WT Allele (LOA) pass 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 47057 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PG00066_X_1_G07 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 47057 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PG00066_X_3_G07 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 47057 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PRPGS00066_B_G07 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 47057 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PG00066_X_4_G07 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 47057 Knockout First, Reporter-tagged insertion with conditional potential (Promotor Driven Cassette) PG00066_X_2_G07 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
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Intermediate Vectors

Design ID Vector Type Intermediate Vector
47057 Knockout First, Reporter-tagged insertion with conditional potential PCS00066_A_G07
47057 Knockout First, Reporter-tagged insertion with conditional potential PCS00066_B_G07