Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI13483
Trapped Gene
Cyp3a13 (ENSMUSG00000029727)
Vector Insertion
Chr 5: 138335593 - 138336410
Public Clones CMHD-GT_277G06-3 (cmhd) CMHD-GT_435B4-3 (cmhd)
Private Clones not available
Severity of mutation (?) Insertion after 94% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000535395 (Chr5:138336411..138336573 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ATTGGCATGAGGTTTGCTCT Chr5:138336481..138336500 59.7 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000535395 (Chr5:138336411..138336573 -)
Downstram Exon
ENSMUSE00000471988 (Chr5:138335405..138335592 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ATTGGCATGAGGTTTGCTCT Chr5:138336481..138336500 59.7 45 TGGTTTCTGGTCCACAGGAT Chr5:138335432..138335451 60.36 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000497844 Chr5:138362670..138362847 GAAAGGTGCAGCACACAAAA Chr5:138362785..138362804 59.89 45
upstream ENSMUSE00000535402 Chr5:138360227..138360320 TTTCTTGGGGACGATTCTTG Chr5:138360238..138360257 60.04 45
upstream ENSMUSE00000192105 Chr5:138358746..138358798 No primer for this exon
upstream ENSMUSE00000535401 Chr5:138356802..138356901 TGCTGGCTATCACAGATCCA Chr5:138356859..138356878 60.38 50
upstream ENSMUSE00000192109 Chr5:138353067..138353180 No primer for this exon
upstream ENSMUSE00000192097 Chr5:138352767..138352855 TGAGGCAGGGATTAGGAGAA Chr5:138352790..138352809 59.77 50
upstream ENSMUSE00000192102 Chr5:138351144..138351292 ATTCCCTCAACAACCCACAG Chr5:138351214..138351233 59.82 50
upstream ENSMUSE00000192099 Chr5:138348662..138348789 TTTCCATTCCTTACCCCAGT Chr5:138348765..138348784 58.36 45
upstream ENSMUSE00000192106 Chr5:138346758..138346824 CAGCTGATGATAAACTCCCAGA Chr5:138346785..138346806 59.34 45.46
upstream ENSMUSE00000192110 Chr5:138341477..138341637 GATGAAATTGATGCGGCTCT Chr5:138341487..138341506 60.19 45
upstream ENSMUSE00000535397 Chr5:138340026..138340252 TGGGCTATTCATTCCCAAAG Chr5:138340108..138340127 59.89 45
upstream ENSMUSE00000535395 Chr5:138336411..138336573 ATTGGCATGAGGTTTGCTCT Chr5:138336481..138336500 59.7 45

*** Putative Vector Insertion (Chr 5: 138335593 - 138336410) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000471988 Chr5:138335405..138335592 TGGTTTCTGGTCCACAGGAT Chr5:138335432..138335451 60.36 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TTTTAATCGCCTTGCAGCAC Chr5:138336342..138336362 61.26 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGGAGTTTTGCTTTTCGTGA Chr5:138336354..138336374 60.23 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000029727