Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI36759
Trapped Gene
Uba3 (ENSMUSG00000030061)
Vector Insertion
Chr 6: 97151910 - 97153589
Public Clones IST14504B1 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 13% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000244911 (Chr6:97153468..97153588 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTCTGGACCCTTCACACACC Chr6:97153491..97153510 60.42 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000244911 (Chr6:97153468..97153588 -)
Downstram Exon
ENSMUSE00000244909 (Chr6:97151911..97151991 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTCTGGACCCTTCACACACC Chr6:97153491..97153510 60.42 60 ACATCCTAAGCCACCAGCTC Chr6:97151907..97151926 59.31 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000695343 Chr6:97155527..97155554 No primer for this exon
upstream ENSMUSE00000244916 Chr6:97155300..97155341 TAGAGGAGCTGCTGGCTGAG Chr6:97155302..97155321 60.97 60
upstream ENSMUSE00000244911 Chr6:97153468..97153588 GTCTGGACCCTTCACACACC Chr6:97153491..97153510 60.42 60
upstream ENSMUSE00000244909 Chr6:97151911..97151991 AGCTGGTGGCTTAGGATGTG Chr6:97151928..97151947 60.28 55

*** Putative Vector Insertion (Chr 6: 97151910 - 97153589) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000244906 Chr6:97149233..97149315 No primer for this exon
downstream ENSMUSE00000194806 Chr6:97146831..97146911 TTGGGTCTTCCGACATCTTT Chr6:97146866..97146885 59.53 45
downstream ENSMUSE00000194801 Chr6:97142735..97142778 No primer for this exon
downstream ENSMUSE00000194788 Chr6:97141520..97141584 GTCCAGGCCACATACAATGA Chr6:97141534..97141553 59.37 50
downstream ENSMUSE00000194797 Chr6:97141087..97141242 GCACTCAATACAAGCGGTCA Chr6:97141089..97141108 59.87 50
downstream ENSMUSE00000194803 Chr6:97139748..97139850 AGGCTGCTCTTTAGGCCATT Chr6:97139733..97139752 60.36 50
downstream ENSMUSE00000194791 Chr6:97139523..97139636 No primer for this exon
downstream ENSMUSE00000194785 Chr6:97136657..97136710 No primer for this exon
downstream ENSMUSE00000194799 Chr6:97136231..97136267 TTGAAAACCTCAGTGGCACA Chr6:97136221..97136240 60.28 45
downstream ENSMUSE00000194793 Chr6:97136063..97136144 CAGCCCATCTACATCATTGAA Chr6:97136071..97136091 58.59 42.86
downstream ENSMUSE00000194787 Chr6:97135644..97135744 TTGGCTACATGCTGGACAGT Chr6:97135699..97135718 59.32 50
downstream ENSMUSE00000194786 Chr6:97135477..97135540 GCTGTGATAGCCGGAGACTT Chr6:97135492..97135511 59.46 55
downstream ENSMUSE00000194795 Chr6:97135335..97135389 GGCCTGGTTCGTTCTTCAAT Chr6:97135336..97135355 61.38 50
downstream ENSMUSE00000695311 Chr6:97133813..97134859 TTCTTGGATGTTCCCCTCTG Chr6:97134646..97134665 60.04 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAGAATGGCTGTTGATGGTG Chr6:97153570..97153590 60.11 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAGAATGGCTGTTGATGGTG Chr6:97153570..97153590 60.11 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000030061