Tools for the biologist enabling optimized use of gene trap clones
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About
UniTrap UNI29721
Trapped Gene
Epas1 (
ENSMUSG00000024140
)
Vector Insertion
Chr 17: 87153649 - 87195014
Public Clones
E092C02 (ggtc)
(ggtc)
IST14673F12 (tigm)
IST14259D4 (tigm)
Private Clones
not available
Severity of mutation
(?)
Insertion after 0% of polypeptide chain
Suggested RT-PCR
(?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000400252 (Chr17:87153482..87153648 +)
Vector Sequence
Forward Primer
Location
Tm
%GC
LacZrt Reverse Primer
Location
CCTGCGAGCACTAAAGACCT
Chr17:87153488..87153507
59.64
55
TGGCGAAAGGGGGATGTG
Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000400252 (Chr17:87153482..87153648 +)
Downstram Exon
ENSMUSE00000691023 (Chr17:87195015..87195036 +)
Forward Primer
Location
Tm
%GC
Reverse Primer
Location
Tm
%GC
CCTGCGAGCACTAAAGACCT
Chr17:87153488..87153507
59.64
55
TTTCTGCCTGTCTTTGAGCA
Chr17:87195038..87195057
59.72
45
Other possible Exon-Primers for RT-PCR
(?)
Exon ID
Exon location
Forward Primer
Primer location
Tm
%GC
upstream
ENSMUSE00000400252
Chr17:87153482..87153648
CCTGCGAGCACTAAAGACCT
Chr17:87153488..87153507
59.64
55
*** Putative Vector Insertion (Chr 17: 87153649 - 87195014) ***
Exon ID
Exon location
Reverse Primer
Primer location
Tm
%GC
downstream
ENSMUSE00000691023
Chr17:87195015..87195036
TTTCTGCCTGTCTTTGAGCA
Chr17:87195038..87195057
59.72
45
downstream
ENSMUSE00000691022
Chr17:87196452..87196646
GGAGCTTATGTGTCCGAAGG
Chr17:87196640..87196659
59.69
55
downstream
ENSMUSE00000242509
Chr17:87196456..87196646
GGAGCTTATGTGTCCGAAGG
Chr17:87196640..87196659
59.69
55
downstream
ENSMUSE00000138650
Chr17:87204533..87204684
TTGCTGATGTTTTCCGACAG
Chr17:87204668..87204687
59.84
45
downstream
ENSMUSE00000138638
Chr17:87205097..87205181
GATCTCCTCATGGTCGCAAG
Chr17:87205159..87205178
60.77
55
downstream
ENSMUSE00000138654
Chr17:87208698..87208816
GGTGGACACGTCTTTGCTCT
Chr17:87208738..87208757
60.31
55
downstream
ENSMUSE00000138643
Chr17:87208924..87209129
ACTTCATGTCCATGCTGTGG
Chr17:87209113..87209132
59.55
50
downstream
ENSMUSE00000242439
Chr17:87217695..87217801
CATAGGCAGAGCGTCCAAGT
Chr17:87217753..87217772
60.42
55
downstream
ENSMUSE00000242428
Chr17:87223020..87223167
CTCCGTGTTTGGCTAGCATC
Chr17:87223079..87223098
60.8
55
downstream
ENSMUSE00000138641
Chr17:87223747..87223961
ACCACGTCGTTCTTCTCGAT
Chr17:87223773..87223792
59.73
50
downstream
ENSMUSE00000138648
Chr17:87224920..87225110
CCATAGGCTGAGGGTTCATC
Chr17:87224956..87224975
59.51
55
downstream
ENSMUSE00000242381
Chr17:87225845..87225955
TGGCGAAAAGCTTCTCAATC
Chr17:87225923..87225942
60.47
45
downstream
ENSMUSE00000242354
Chr17:87226832..87227337
TAGGGATGTAGGGTGCCAAG
Chr17:87226883..87226902
59.95
55
downstream
ENSMUSE00000242323
Chr17:87228030..87228156
CCGCTTTAGCTTCAGCTTGT
Chr17:87228117..87228136
59.79
50
downstream
ENSMUSE00000138646
Chr17:87228326..87228440
TTCGCACTGATGGTCTTGTC
Chr17:87228432..87228451
59.84
50
downstream
ENSMUSE00000242272
Chr17:87228650..87228823
AGCTCCTGGAGGACCGTAGT
Chr17:87228813..87228832
60.27
60
downstream
ENSMUSE00000481232
Chr17:87230291..87230921
TGAGTTGAATGACGCCAAAG
Chr17:87230841..87230860
59.84
45
downstream
ENSMUSE00000691024
Chr17:87230291..87230659
CAGTTCCGGCAACAGGTAAG
Chr17:87230349..87230368
60.68
55
Suggested Genomic-PCR
(?)
Forward
Forward location
Tm
%GC
Reverse name
Reverse
Reverse Location
PCR 1
GGTACCCCCTTTTGTCCCTA
Chr17:87177607..87177627
60.05
55
L232
GATGTGCTGCAAGGCGATTA
Gene trap vector
PCR 2
GGTACCCCCTTTTGTCCCTA
Chr17:87177607..87177627
60.05
55
LacZ
CCAGGGTTTTCCCAGTCACG
Gene trap vector
Come back to gene
ENSMUSG00000024140