Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI20101
Trapped Gene
Zfyve9 (ENSMUSG00000034557)
Vector Insertion
Chr 4: 108357637 - 108361753
Public Clones not available
Private Clones OST372966 (lexicon) OST361368 (lexicon) OST356405 (lexicon) OST24562 (lexicon)
OST19150 (lexicon) OST10391 (lexicon)
Severity of mutation (?) Insertion after 62% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000631541 (Chr4:108361754..108361874 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGTTGGAAGTCCTGTTGGAA Chr4:108361820..108361839 59.94 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000631541 (Chr4:108361754..108361874 -)
Downstram Exon
ENSMUSE00000631540 (Chr4:108357514..108357636 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGTTGGAAGTCCTGTTGGAA Chr4:108361820..108361839 59.94 50 TTGCTGCATGACTGAAATCTG Chr4:108357568..108357588 60 42.86

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000670776 Chr4:108453058..108453403 GCTCTCGGCTGAGTGACGAC Chr4:108453078..108453097 63.68 65
upstream ENSMUSE00000670782 Chr4:108453058..108453108 GCTCTCGGCTGAGTGACGAC Chr4:108453078..108453097 63.68 65
upstream ENSMUSE00000670781 Chr4:108400061..108400150 AGCAAGCACAGAGAAGACTGC Chr4:108400073..108400093 59.95 52.38
upstream ENSMUSE00000670780 Chr4:108396412..108396517 CCAAGCAGAGGCTTACAACC Chr4:108396448..108396467 59.88 55
upstream ENSMUSE00000712012 Chr4:108396412..108396481 CCAAGCAGAGGCTTACAACC Chr4:108396448..108396467 59.88 55
upstream ENSMUSE00000718549 Chr4:108396412..108396481 CCAAGCAGAGGCTTACAACC Chr4:108396448..108396467 59.88 55
upstream ENSMUSE00000670793 Chr4:108390394..108392417 ATCTCTGTGCGGCTTCATCT Chr4:108391789..108391808 59.98 50
upstream ENSMUSE00000670791 Chr4:108368374..108368473 TTCCTGCTGCAGCCTAAAAT Chr4:108368443..108368462 59.98 45
upstream ENSMUSE00000631543 Chr4:108365925..108366101 AAGCCAGAGCCCTAACCCTA Chr4:108366051..108366070 60.22 55
upstream ENSMUSE00000631542 Chr4:108364227..108364396 TTGGTTTGCTGATGGGATCT Chr4:108364349..108364368 60.46 45
upstream ENSMUSE00000631541 Chr4:108361754..108361874 GGTTGGAAGTCCTGTTGGAA Chr4:108361820..108361839 59.94 50

*** Putative Vector Insertion (Chr 4: 108357637 - 108361753) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000631540 Chr4:108357514..108357636 TTGCTGCATGACTGAAATCTG Chr4:108357568..108357588 60 42.86
downstream ENSMUSE00000631539 Chr4:108354739..108354894 CATGCATCCCTTTGGTTGTA Chr4:108354828..108354847 59.4 45
downstream ENSMUSE00000631538 Chr4:108353507..108353731 AACAGGTAAGGAGGGGTTGG Chr4:108353571..108353590 60.22 55
downstream ENSMUSE00000631537 Chr4:108350315..108350397 ATGGCTTCCGAAATCTGACA Chr4:108350334..108350353 60.6 45
downstream ENSMUSE00000631536 Chr4:108347058..108347162 ACCTTGGACAACTGGCAGAG Chr4:108347099..108347118 60.3 55
downstream ENSMUSE00000269658 Chr4:108333170..108333320 TCTGCCTTTTCATTGAAGCA Chr4:108333228..108333247 59.54 40
downstream ENSMUSE00000269651 Chr4:108329532..108329612 CTGGACTTGGCAAGGTATCC Chr4:108329521..108329540 59.55 55
downstream ENSMUSE00000269644 Chr4:108322789..108322951 GAGTCCATGTTCTCCGCAGT Chr4:108322890..108322909 60.27 55
downstream ENSMUSE00000269636 Chr4:108316941..108317046 TCTGATGACTTTTCCGTTTGC Chr4:108316928..108316948 60.24 42.86
downstream ENSMUSE00000269625 Chr4:108314693..108314869 GTCACACGGAGTCCCAGTTT Chr4:108314687..108314706 60.01 55
downstream ENSMUSE00000707976 Chr4:108311864..108312025 AGGCTCCTCCATGAATGACA Chr4:108311907..108311926 60.62 50
downstream ENSMUSE00000708983 Chr4:108311864..108312025 AGGCTCCTCCATGAATGACA Chr4:108311907..108311926 60.62 50
downstream ENSMUSE00000670779 Chr4:108310071..108312025 TGCAGGCAAGAAAACATCAG Chr4:108310510..108310529 59.99 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AAGGAGGTATGTGGGCTTCA Chr4:108358738..108358758 59.55 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AAGGAGGTATGTGGGCTTCA Chr4:108358738..108358758 59.55 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000034557