IKMC Project Report - EUCOMM (ID: 77056)

Cyp20a1 MGI:1925201 ENSMUSG00000049439 OTTMUSG00000021516
Pre-pipeline Designs Vectors ES Cells Mice
Mice - Phenotype Data Available

Mice

Microinjection Status Allele Allele Type ES Cell Clone Genetic Background QC Data MI/Distribution Centre
contact distributor Genotype confirmed Cyp20a1tm1a(EUCOMM)Wtsi Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) EPD0535_1_F10 C57BL/6NTac;C57BL/6NTac;C57BL/6N-Atm1Brd/a view
( about ) 		WTSI/Oulu
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 pass 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 3 wild type transcripts, of which 2 are protein-coding. Following removal of the floxed region, 2 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'. 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
ENSMUST00000060608 protein_coding No protein product (NMD) view

Predicted translation: 

MLDFAIFAVTFLLALVGAVLYLYPASRQASGIPGLTPTEEKDGNLPDIVNSGSLHEFLVNLHERYGPVVSFWFGRRLVVS
LGTTDVLKQHFNPNKTSFRRIVR*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00001089881 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00001067903 CDS CDS
ENSMUSE00000415874 Cyt_P450 (54/403 aa) CDS CDS
ENSMUSE00001016029 Cyt_P450 (48/403 aa) CDS Deleted
ENSMUSE00001038543 Cyt_P450 (56/403 aa) CDS CDS + stop codon + UTR
ENSMUSE00001025966 Cyt_P450 (27/403 aa) CDS
ENSMUSE00001031415 Cyt_P450 (39/403 aa) CDS
ENSMUSE00001086001 Cyt_P450 (19/403 aa) CDS
ENSMUSE00001006308 Cyt_P450 (41/403 aa) CDS
ENSMUSE00001078224 Cyt_P450 (38/403 aa) CDS
ENSMUSE00001069004 Cyt_P450 (22/403 aa) CDS
ENSMUSE00001010657 Cyt_P450 (31/403 aa) CDS
ENSMUSE00001092234 Cyt_P450 (34/403 aa) CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000114202 protein_coding No protein product (NMD) view

Predicted translation: 

MLDFAIFAVTFLLALVGAVLYLYPASRQASGIPGLTPTEEKDGNLPDIVNSGSLHEFLVNLHERYGPVVSFWFGRRLVVS
LGTTDVLKQHFNPNKTSFRRIVR*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00001089881 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00001067903 CDS CDS
ENSMUSE00000415874 Cyt_P450 (54/223 aa) CDS CDS
ENSMUSE00001016029 Cyt_P450 (48/223 aa) CDS Deleted
ENSMUSE00001038543 Cyt_P450 (56/223 aa) CDS CDS + stop codon + UTR
ENSMUSE00001025966 Cyt_P450 (27/223 aa) CDS
ENSMUSE00001031415 Cyt_P450 (39/223 aa) CDS
ENSMUSE00000698492 Cyt_P450 (1/223 aa) CDS + stop codon + UTR
Legend: in frame deleted frameshifted
ENSMUST00000148443 nonsense_mediated_decay No protein product (NMD) view

Predicted translation: 

MLDFAIFAVTFLLALVGAVLYLYPASRQASGIPGLTPTEENFQKNC*

Ensembl Exon ID Pfam Domains WildType Floxed
ENSMUSE00001089881 UTR + start codon + CDS UTR + start codon + CDS
ENSMUSE00001067903 CDS CDS
ENSMUSE00000977753 CDS + stop codon + UTR Deleted
ENSMUSE00001028173 UTR CDS + stop codon + UTR
ENSMUSE00001069970 UTR UTR
ENSMUSE00001041701 UTR UTR
ENSMUSE00001016401 UTR UTR
ENSMUSE00001003161 UTR UTR
ENSMUSE00000996379 UTR UTR
ENSMUSE00001068020 UTR UTR
ENSMUSE00001043482 UTR UTR
ENSMUSE00001062171 UTR UTR
Legend: in frame deleted frameshifted
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ES Cell Clones 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 EPD0535_1_F10 HTGR04003_Z_5_D03 Cyp20a1tm1a(EUCOMM)Wtsi Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) JM8A3.N1 view yes view    ( about )
Production Centre
5' Screen pass 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 pass 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 EPD0535_1_E09 HTGR04003_Z_5_D03 Cyp20a1tm1a(EUCOMM)Wtsi Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen not confirmed 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 EPD0535_1_E12 HTGR04003_Z_5_D03 Cyp20a1tm1a(EUCOMM)Wtsi Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen not confirmed 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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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 EPD0535_1_B11 HTGR04003_Z_5_D03 Cyp20a1tm1e(EUCOMM)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen not confirmed 3' Screen not confirmed
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 EPD0535_1_B12 HTGR04003_Z_5_D03 Cyp20a1tm1e(EUCOMM)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen not confirmed 3' Screen not confirmed
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 EPD0535_1_C09 HTGR04003_Z_5_D03 Cyp20a1tm1e(EUCOMM)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen not confirmed 3' Screen not confirmed
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 EPD0535_1_C10 HTGR04003_Z_5_D03 Cyp20a1tm1e(EUCOMM)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen not confirmed 3' Screen not confirmed
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 EPD0535_1_D09 HTGR04003_Z_5_D03 Cyp20a1tm1e(EUCOMM)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen not confirmed 3' Screen not confirmed
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 EPD0535_1_D10 HTGR04003_Z_5_D03 Cyp20a1tm1e(EUCOMM)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen not confirmed 3' Screen not confirmed
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 EPD0535_1_D12 HTGR04003_Z_5_D03 Cyp20a1tm1e(EUCOMM)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen not confirmed 3' Screen not confirmed
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 EPD0535_1_F12 HTGR04003_Z_5_D03 Cyp20a1tm1e(EUCOMM)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen not confirmed 3' Screen not confirmed
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 EPD0535_1_G09 HTGR04003_Z_5_D03 Cyp20a1tm1e(EUCOMM)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen not confirmed 3' Screen not confirmed
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 EPD0535_1_H09 HTGR04003_Z_5_D03 Cyp20a1tm1e(EUCOMM)Wtsi Targeted Non-Conditional (Promotor Driven Cassette) JM8A3.N1 view no view    ( about )
Production Centre
5' Screen pass LoxP Screen not confirmed 3' Screen not confirmed
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 49437 Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) HTGR04003_Z_5_A12 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 49437 Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) HTGR04003_Z_3_B06 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 49437 Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) HTGR04003_Z_5_G02 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 49437 Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) HTGR04003_Z_5_D03 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 49437 Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) HTGR04003_Z_3_G08 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 49437 Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) HTGR04003_Z_5_G07 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 49437 Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) HTGR04003_Z_5_C08 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 49437 Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) HTGR04003_Z_3_C11 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 49437 Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) HTGR04003_Z_3_D04 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
order 49437 Knockout-First - Reporter Tagged Insertion (Promotor Driven Cassette) HTGR04003_Z_3_G02 L1L2_Bact_P L3L4_pD223_DTA_T_spec view
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Intermediate Vectors

Design ID Vector Type Intermediate Vector
49437 Knockout-First - Reporter Tagged Insertion PCS04003_A_C12