Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI3810
Trapped Gene
Eif3b (ENSMUSG00000056076)
Vector Insertion
Chr 5: 140917066 - 140917690
Public Clones CD0145 (sanger)
Private Clones not available
Severity of mutation (?) Insertion after 73% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000504465 (Chr5:140916940..140917065 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AACAACAAGGACCGCTTCTG Chr5:140916991..140917010 60.29 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000504465 (Chr5:140916940..140917065 +)
Downstram Exon
ENSMUSE00000503205 (Chr5:140917691..140917768 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AACAACAAGGACCGCTTCTG Chr5:140916991..140917010 60.29 50 AAGCGATCCTTCTGCTCAAA Chr5:140917749..140917768 60.1 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000462872 Chr5:140895259..140895780 ACCCCGAGGATTTCGTAGAC Chr5:140895745..140895764 60.33 55
upstream ENSMUSE00000706481 Chr5:140895315..140895780 ACCCCGAGGATTTCGTAGAC Chr5:140895745..140895764 60.33 55
upstream ENSMUSE00000463882 Chr5:140901216..140901408 GCCAGACCGACTTGAGAAAC Chr5:140901304..140901323 59.85 55
upstream ENSMUSE00000461089 Chr5:140902391..140902510 GGTCAACCTCTTCACCGACT Chr5:140902484..140902503 59.15 55
upstream ENSMUSE00000462079 Chr5:140902737..140902794 ATCAGCGATGAGTGGGACAT Chr5:140902747..140902766 60.51 50
upstream ENSMUSE00000466462 Chr5:140902971..140903099 ATTTGCGTTACTGGCTGGAA Chr5:140902975..140902994 60.64 45
upstream ENSMUSE00000467549 Chr5:140903364..140903521 AGGGGACAAGTTCAAGCAGA Chr5:140903447..140903466 59.84 50
upstream ENSMUSE00000464703 Chr5:140906006..140906137 CTCACGGGCCATAAGAAGAG Chr5:140906076..140906095 59.83 55
upstream ENSMUSE00000465740 Chr5:140906216..140906282 TGCAAGAATGACCCTGGATA Chr5:140906240..140906259 59.08 45
upstream ENSMUSE00000495070 Chr5:140906925..140906971 GGCCTTTTGGACAAGAAGAGTT Chr5:140906931..140906952 60.97 45.46
upstream ENSMUSE00000495987 Chr5:140908878..140909088 TCTCCGGGTGGTAACATCAT Chr5:140908891..140908910 60.19 50
upstream ENSMUSE00000500987 Chr5:140912202..140912274 ATTCCGAATGAGGGAGAAGC Chr5:140912225..140912244 60.55 50
upstream ENSMUSE00000501538 Chr5:140913239..140913361 AGAGCAACGGGAAGATTGAA Chr5:140913335..140913354 59.81 45
upstream ENSMUSE00000498625 Chr5:140914270..140914348 GCAAGCCAACACCATCTTCT Chr5:140914286..140914305 60.26 50
upstream ENSMUSE00000499766 Chr5:140915867..140916005 GCGTTTGTCGACACTTCAGA Chr5:140915883..140915902 60.03 50
upstream ENSMUSE00000504465 Chr5:140916940..140917065 AACAACAAGGACCGCTTCTG Chr5:140916991..140917010 60.29 50

*** Putative Vector Insertion (Chr 5: 140917066 - 140917690) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000503205 Chr5:140917691..140917768 AAGCGATCCTTCTGCTCAAA Chr5:140917749..140917768 60.1 45
downstream ENSMUSE00000504091 Chr5:140918078..140918186 AGACGCTCGTTCTTCTGCTT Chr5:140918175..140918194 59.38 50
downstream ENSMUSE00000477070 Chr5:140918274..140918335 CTGTCCAGCTCGTCAGTGTC Chr5:140918301..140918320 59.6 60
downstream ENSMUSE00000648337 Chr5:140918274..140918390 AGGAATGACTTCCTCGGTGA Chr5:140918362..140918381 59.65 50
downstream ENSMUSE00000481881 Chr5:140918338..140918390 AGGAATGACTTCCTCGGTGA Chr5:140918362..140918381 59.65 50
downstream ENSMUSE00000482849 Chr5:140918828..140919314 GGAACAGCAAGTCCGTGAAT Chr5:140919160..140919179 60.12 50
downstream ENSMUSE00000648336 Chr5:140918828..140919312 GGAACAGCAAGTCCGTGAAT Chr5:140919160..140919179 60.12 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ACTCCTCAGCCAGGATCAGA Chr5:140917042..140917062 59.94 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ACTCCTCAGCCAGGATCAGA Chr5:140917042..140917062 59.94 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000056076