Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI1717
Trapped Gene
Agpat4 (ENSMUSG00000023827)
Vector Insertion
Chr 17: 12391788 - 12403076
Public Clones CG0722 (sanger) RRF444 (baygenomics) RRF360 (baygenomics)
Private Clones OST59808 (lexicon)
Severity of mutation (?) Insertion after 31% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000371130 (Chr17:12391618..12391787 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TCGTGGTCCTCAATCACAAG Chr17:12391711..12391730 59.68 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000371130 (Chr17:12391618..12391787 +)
Downstram Exon
ENSMUSE00000135585 (Chr17:12403077..12403238 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TCGTGGTCCTCAATCACAAG Chr17:12391711..12391730 59.68 50 TCCACGAAGTACCACATCCA Chr17:12403150..12403169 59.96 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000336429 Chr17:12312150..12312281 TTGTGAGCACCAATGCAAAG Chr17:12312246..12312265 60.84 45
upstream ENSMUSE00000377093 Chr17:12344558..12344815 AGCTGTTCCGCAAGATCAAT Chr17:12344768..12344787 59.84 45
upstream ENSMUSE00000371130 Chr17:12391618..12391787 TCGTGGTCCTCAATCACAAG Chr17:12391711..12391730 59.68 50

*** Putative Vector Insertion (Chr 17: 12391788 - 12403076) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000135585 Chr17:12403077..12403238 TCCACGAAGTACCACATCCA Chr17:12403150..12403169 59.96 50
downstream ENSMUSE00000135584 Chr17:12403505..12403658 CAACATCTCGCAAGCACTTC Chr17:12403661..12403680 59.6 50
downstream ENSMUSE00000135586 Chr17:12406321..12406423 AGCAGTGTTGGGTTTTCATTG Chr17:12406380..12406400 60.02 42.86
downstream ENSMUSE00000556040 Chr17:12408142..12408217 TGTGTAACCAGGCAGAGCAC Chr17:12408203..12408222 59.9 55
downstream ENSMUSE00000135582 Chr17:12409560..12409758 CCAGAACAACCAGTTGACCA Chr17:12409661..12409680 59.56 50
downstream ENSMUSE00000389483 Chr17:12411851..12412506 CAGTCCGTTTGTTTCCGTTT Chr17:12411947..12411966 60.01 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TTGAAACTGGGGCCTAATTC Chr17:12400789..12400809 59.02 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TAAGTGGGCTGTGCCTTCAC Chr17:12400790..12400810 61.24 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000023827