Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI25684
Trapped Gene
Eef2k (ENSMUSG00000035064)
Vector Insertion
Chr 7: 127994884 - 128001857
Public Clones not available
Private Clones OST170518 (lexicon)
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000670167 (Chr7:127994715..127994883 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGTCACAAGGTAGGCAAGGA Chr7:127994792..127994811 60.11 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000670167 (Chr7:127994715..127994883 +)
Downstram Exon
ENSMUSE00000716617 (Chr7:128001858..128002176 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGTCACAAGGTAGGCAAGGA Chr7:127994792..127994811 60.11 55 AGCCGCACTCTGTTTTTGTT Chr7:128002144..128002163 59.92 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000463208 Chr7:127986369..127986614 TTTGTACCGGGGATTCTCCT Chr7:127986494..127986513 60.68 50
upstream ENSMUSE00000670168 Chr7:127987114..127987299 GGGGAGTATTTCTGCGTCCT Chr7:127987185..127987204 60.46 55
upstream ENSMUSE00000670164 Chr7:127994703..127994883 GGTCACAAGGTAGGCAAGGA Chr7:127994792..127994811 60.11 55
upstream ENSMUSE00000670167 Chr7:127994715..127994883 GGTCACAAGGTAGGCAAGGA Chr7:127994792..127994811 60.11 55

*** Putative Vector Insertion (Chr 7: 127994884 - 128001857) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000261270 Chr7:128001858..128002176 AGCCGCACTCTGTTTTTGTT Chr7:128002144..128002163 59.92 45
downstream ENSMUSE00000716617 Chr7:128001858..128002176 AGCCGCACTCTGTTTTTGTT Chr7:128002144..128002163 59.92 45
downstream ENSMUSE00000261263 Chr7:128016823..128016923 ATGTGCTTGGCTTTCTCGAT Chr7:128016863..128016882 59.84 45
downstream ENSMUSE00000261256 Chr7:128019946..128020006 ACGCCATCTTGATCAGAACC Chr7:128020004..128020023 60.08 50
downstream ENSMUSE00000261249 Chr7:128023328..128023365 GTCCTGAAGCACTCCCTCAT Chr7:128023368..128023387 59.26 55
downstream ENSMUSE00000261243 Chr7:128023819..128023990 GGAGCTGCACATCCTCAAAG Chr7:128023937..128023956 60.94 55
downstream ENSMUSE00000261239 Chr7:128028172..128028321 TGTGGGGTTAGTCGGATGTT Chr7:128028323..128028342 60.23 50
downstream ENSMUSE00000261230 Chr7:128028936..128029068 CTGATGACCAGAACGCTCAA Chr7:128028977..128028996 59.98 50
downstream ENSMUSE00000261220 Chr7:128029324..128029451 CTGATTCACCGCATCCTGTT Chr7:128029439..128029458 61.07 50
downstream ENSMUSE00000632652 Chr7:128029324..128029400 GGTTGCAGGCATGAGAGTAGA Chr7:128029370..128029390 60.41 52.38
downstream ENSMUSE00000670163 Chr7:128029324..128029399 GGTTGCAGGCATGAGAGTAGA Chr7:128029370..128029390 60.41 52.38
downstream ENSMUSE00000261211 Chr7:128030100..128030301 TTCTGGCTGTGTGGTGTAGC Chr7:128030290..128030309 59.9 55
downstream ENSMUSE00000261204 Chr7:128031373..128031440 GCCCATGTTATCGAGGTCAC Chr7:128031410..128031429 60.35 55
downstream ENSMUSE00000632650 Chr7:128031389..128031440 CTCTAGGGCCCATGTTATCG Chr7:128031417..128031436 59.55 55
downstream ENSMUSE00000670165 Chr7:128031390..128031440 ATGCGGTCATGGTCTCTAGG Chr7:128031430..128031449 60.1 55
downstream ENSMUSE00000261198 Chr7:128032704..128032781 CTCGCTTGGATACCCACTGT Chr7:128032745..128032764 60.13 55
downstream ENSMUSE00000261191 Chr7:128033203..128033265 CCAGGCTATCCTCGTCAGAT Chr7:128033254..128033273 59.26 55
downstream ENSMUSE00000261184 Chr7:128035110..128035244 GACTTCCCGATTTTCCTTCC Chr7:128035234..128035253 59.88 50
downstream ENSMUSE00000261177 Chr7:128035379..128035567 GCTGACTCTCGATCCCACTC Chr7:128035458..128035477 59.95 60
downstream ENSMUSE00000261172 Chr7:128038698..128038822 TTCTGCCGCCTTCAGTAAGT Chr7:128038754..128038773 60.01 50
downstream ENSMUSE00000261164 Chr7:128042882..128043060 CTCGTCCTGTATCCCATCGT Chr7:128042981..128043000 59.95 55
downstream ENSMUSE00000632649 Chr7:128043775..128043824 TGAGGTCCACTGGCCTACAT Chr7:128043809..128043828 60.53 55
downstream ENSMUSE00000670162 Chr7:128043775..128043886 GCAGTCAAAAGGATCCAAGG Chr7:128043853..128043872 59.67 50
downstream ENSMUSE00000423966 Chr7:128046796..128047508 CCGCCTTCTCGTAGTACTGG Chr7:128046879..128046898 59.89 60
downstream ENSMUSE00000670166 Chr7:128046796..128047010 CCGCCTTCTCGTAGTACTGG Chr7:128046879..128046898 59.89 60
downstream ENSMUSE00000705530 Chr7:128046796..128047011 CCGCCTTCTCGTAGTACTGG Chr7:128046879..128046898 59.89 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATGCACTACCTGGCCTGACT Chr7:127997874..127997894 59.75 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ATGCACTACCTGGCCTGACT Chr7:127997874..127997894 59.75 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000035064