Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI221
Trapped Gene
Kif22 (ENSMUSG00000030677)
Vector Insertion
Chr 7: 134179455 - 134185857
Public Clones GC0658 (tigem) (sanger) SYA337 (baygenomics) RHA173 (baygenomics) D101C12 (ggtc)
M045C05 (ggtc) D146C06 (ggtc) CMHD-GT_510G12-3 (cmhd) IST13054H4 (tigm)
IST14694F10 (tigm)
Private Clones OST67547 (lexicon)
Severity of mutation (?) Insertion after 3% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000201894 (Chr7:134185858..134185934 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CCGAAGAAGGGAAGGCTATG Chr7:134185913..134185932 61.07 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000201894 (Chr7:134185858..134185934 -)
Downstram Exon
ENSMUSE00000201906 (Chr7:134179262..134179454 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CCGAAGAAGGGAAGGCTATG Chr7:134185913..134185932 61.07 55 AGCCCTCCTTTGCTTACACA Chr7:134179402..134179421 59.88 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000201894 Chr7:134185858..134185934 CCGAAGAAGGGAAGGCTATG Chr7:134185913..134185932 61.07 55

*** Putative Vector Insertion (Chr 7: 134179455 - 134185857) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000201906 Chr7:134179262..134179454 AGCCCTCCTTTGCTTACACA Chr7:134179402..134179421 59.88 50
downstream ENSMUSE00000201900 Chr7:134177638..134177765 TGAGTGCTCTTCTCGCCATA Chr7:134177711..134177730 59.7 50
downstream ENSMUSE00000201897 Chr7:134177392..134177546 AGTTGCAGGAGGTCCATGAG Chr7:134177449..134177468 60.26 55
downstream ENSMUSE00000201896 Chr7:134176983..134177192 TGGCTCCTACAGCTCGATTT Chr7:134177011..134177030 59.98 50
downstream ENSMUSE00000201902 Chr7:134176676..134176906 TTGCTGTCCCGGTATGGTAT Chr7:134176673..134176692 60.21 50
downstream ENSMUSE00000201905 Chr7:134176396..134176549 TGTTGGCAATGAGGATGCTA Chr7:134176485..134176504 60.22 45
downstream ENSMUSE00000669477 Chr7:134176148..134176278 AGGGCTCCCAGTTGATTCTT Chr7:134176162..134176181 60.07 50
downstream ENSMUSE00000201895 Chr7:134176143..134176278 AGGGCTCCCAGTTGATTCTT Chr7:134176162..134176181 60.07 50
downstream ENSMUSE00000201893 Chr7:134174443..134174611 CCCAGCAAACGTTCCATACT Chr7:134174518..134174537 59.99 50
downstream ENSMUSE00000669476 Chr7:134174443..134174616 CCCAGCAAACGTTCCATACT Chr7:134174518..134174537 59.99 50
downstream ENSMUSE00000201903 Chr7:134172849..134173014 CAACACCTTGGCTTCCAGTT Chr7:134172951..134172970 60.15 50
downstream ENSMUSE00000201899 Chr7:134172429..134172490 GTTTGAGGACTCTCGCTGCT Chr7:134172443..134172462 59.75 55
downstream ENSMUSE00000201904 Chr7:134171584..134171796 GCCCATGTGCCAGTAACTCT Chr7:134171687..134171706 60.14 55
downstream ENSMUSE00000201901 Chr7:134171445..134171504 CAGAGATGCCTTCCACCTGT Chr7:134171449..134171468 60.26 55
downstream ENSMUSE00000352471 Chr7:134171246..134171354 No primer for this exon

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CACCACTGCGTGAGACTTACA Chr7:134179812..134179833 59.95 52.38 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CACCACTGCGTGAGACTTACA Chr7:134179812..134179833 59.95 52.38 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 GCTAAGACGTGTCCGCAGTA Chr7:134179882..134179902 59.1 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 CGAAGAAGGGAAGGCTATGA Chr7:134179910..134179930 59.41 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000030677