Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI13482
Trapped Gene
Fbxw7 (ENSMUSG00000028086)
Vector Insertion
Chr 3: 84778551 - 84780095
Public Clones CMHD-GT_286F06-3 (cmhd)
Private Clones not available
Severity of mutation (?) Insertion after 75% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000175995 (Chr3:84778325..84778550 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGCCAGTGTTTACACGTCCT Chr3:84778377..84778396 60.03 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000175995 (Chr3:84778325..84778550 +)
Downstram Exon
ENSMUSE00000175999 (Chr3:84780096..84780306 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGCCAGTGTTTACACGTCCT Chr3:84778377..84778396 60.03 55 GGTGTCCTGTTAGCGTGTGA Chr3:84780195..84780214 59.75 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000674115 Chr3:84619190..84619691 TTCGAGCTTTCTGATCGTGA Chr3:84619404..84619423 59.67 45
upstream ENSMUSE00000674100 Chr3:84619499..84619691 TAGCACCGCTTCTTCCTCAG Chr3:84619577..84619596 60.67 55
upstream ENSMUSE00000674114 Chr3:84667886..84667933 No primer for this exon
upstream ENSMUSE00000674113 Chr3:84668059..84668122 No primer for this exon
upstream ENSMUSE00000674102 Chr3:84668067..84668122 No primer for this exon
upstream ENSMUSE00000674101 Chr3:84694017..84694174 GCACATGGAAACCAGAGACC Chr3:84694119..84694138 60.52 55
upstream ENSMUSE00000674111 Chr3:84707423..84708001 TGGAGACCAGAACGATACCC Chr3:84707689..84707708 59.93 55
upstream ENSMUSE00000720664 Chr3:84707423..84708001 TGGAGACCAGAACGATACCC Chr3:84707689..84707708 59.93 55
upstream ENSMUSE00000674095 Chr3:84729402..84729721 GGTCTGCCTTTTTGTCGAAG Chr3:84729698..84729717 59.85 50
upstream ENSMUSE00000299439 Chr3:84756295..84756561 ATGGGTTTCTACGGCACATT Chr3:84756523..84756542 59.32 45
upstream ENSMUSE00000175994 Chr3:84758797..84758879 GGGAAAGAAACCATGCAAAG Chr3:84758844..84758863 59.55 45
upstream ENSMUSE00000175997 Chr3:84762426..84762567 GCCTTGTACCATGTTCAGCA Chr3:84762434..84762453 59.72 50
upstream ENSMUSE00000175996 Chr3:84769170..84769304 GTGATAGAGCCCCAGTTCCA Chr3:84769254..84769273 60.07 55
upstream ENSMUSE00000176000 Chr3:84771362..84771485 CTGGAGAATTTTGGCTGAGG Chr3:84771430..84771449 59.81 50
upstream ENSMUSE00000674099 Chr3:84771362..84771535 CTGGAGAATTTTGGCTGAGG Chr3:84771430..84771449 59.81 50
upstream ENSMUSE00000176001 Chr3:84773087..84773223 ATGAACCGTTGCACATCAAG Chr3:84773093..84773112 59.57 45
upstream ENSMUSE00000175992 Chr3:84774718..84774831 TTGTGGCAACCGCATAGTTA Chr3:84774762..84774781 60.13 45
upstream ENSMUSE00000175991 Chr3:84776297..84776478 TGGGACATACAGGTGGAGTG Chr3:84776313..84776332 59.39 55
upstream ENSMUSE00000175995 Chr3:84778325..84778550 GGCCAGTGTTTACACGTCCT Chr3:84778377..84778396 60.03 55

*** Putative Vector Insertion (Chr 3: 84778551 - 84780095) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000175999 Chr3:84780096..84780306 GGTGTCCTGTTAGCGTGTGA Chr3:84780195..84780214 59.75 55
downstream ENSMUSE00000590321 Chr3:84781195..84783119 GATGCTACGACTGACCAGCA Chr3:84782710..84782729 60.02 55
downstream ENSMUSE00000674103 Chr3:84781195..84783120 GATGCTACGACTGACCAGCA Chr3:84782710..84782729 60.02 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTGTCTCTCCCACCCTTTGA Chr3:84778562..84778582 60.09 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTGTCTCTCCCACCCTTTGA Chr3:84778562..84778582 60.09 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000028086