Tools for the biologist enabling optimized use of gene trap clones
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UniTrap UNI28108
Trapped Gene 
Phrf1 (ENSMUSG00000038611)
Vector Insertion 
Chr 7: 148442579 - 148442701
Public Clones not available
Private Clones OST39915 (lexicon)
Severity of mutation (?) Insertion after 27% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000668321 (Chr7:148442397..148442578 +)		 Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTTCTCGAATTGCACGAACA Chr7:148442413..148442432 59.85 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000668321 (Chr7:148442397..148442578 +)		 Downstram Exon
ENSMUSE00000586970 (Chr7:148442702..148442821 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTTCTCGAATTGCACGAACA Chr7:148442413..148442432 59.85 45 GGGACAGCACTCTCCTCTTG Chr7:148442766..148442785 59.99 60

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000668320 Chr7:148414683..148414790 GACAGAGGCGGGATCGTCTA Chr7:148414742..148414761 62.65 60
upstream ENSMUSE00000668331 Chr7:148414687..148414790 GACAGAGGCGGGATCGTCTA Chr7:148414742..148414761 62.65 60
upstream ENSMUSE00000713133 Chr7:148417126..148417241 CATGGATGACGACAACTTGG Chr7:148417147..148417166 59.96 50
upstream ENSMUSE00000719095 Chr7:148417126..148417241 CATGGATGACGACAACTTGG Chr7:148417147..148417166 59.96 50
upstream ENSMUSE00000668318 Chr7:148418318..148418440 CAGAGGGGCTGTCTGAAGAG Chr7:148418399..148418418 60.13 60
upstream ENSMUSE00000668329 Chr7:148418318..148418440 CAGAGGGGCTGTCTGAAGAG Chr7:148418399..148418418 60.13 60
upstream ENSMUSE00000668317 Chr7:148423396..148423601 CCTGTGCCCATTATTTCTGC Chr7:148423555..148423574 60.47 50
upstream ENSMUSE00000668328 Chr7:148423396..148423601 CCTGTGCCCATTATTTCTGC Chr7:148423555..148423574 60.47 50
upstream ENSMUSE00000668316 Chr7:148426784..148426867 TTCGTGCCCAGTTTAATGGT Chr7:148426833..148426852 60.37 45
upstream ENSMUSE00000668327 Chr7:148426784..148426867 TTCGTGCCCAGTTTAATGGT Chr7:148426833..148426852 60.37 45
upstream ENSMUSE00000711638 Chr7:148429398..148429795 ACCCGACTTTTTGTGAGGTG Chr7:148429720..148429739 60.01 50
upstream ENSMUSE00000668315 Chr7:148429677..148429795 ACCCGACTTTTTGTGAGGTG Chr7:148429720..148429739 60.01 50
upstream ENSMUSE00000668326 Chr7:148429677..148429795 ACCCGACTTTTTGTGAGGTG Chr7:148429720..148429739 60.01 50
upstream ENSMUSE00000668325 Chr7:148432746..148432843 AGAGTGTACGGTCCCTGGTG Chr7:148432809..148432828 60.03 60
upstream ENSMUSE00000668324 Chr7:148433120..148433295 CACGGCAGAGTGAGAGAGTG Chr7:148433228..148433247 59.76 60
upstream ENSMUSE00000668323 Chr7:148434279..148434408 ATACCAGCGTCCCTTGACAC Chr7:148434356..148434375 60 55
upstream ENSMUSE00000668322 Chr7:148440763..148440890 GTGGGAGTACGAGAGCCAAG Chr7:148440826..148440845 59.87 60
upstream ENSMUSE00000668321 Chr7:148442397..148442578 GTTCTCGAATTGCACGAACA Chr7:148442413..148442432 59.85 45

*** Putative Vector Insertion (Chr 7: 148442579 - 148442701) ***
  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000586970 Chr7:148442702..148442821 GGGACAGCACTCTCCTCTTG Chr7:148442766..148442785 59.99 60
downstream ENSMUSE00000586969 Chr7:148443243..148443397 CCCGGTGTATGACAACATCA Chr7:148443373..148443392 60.24 50
downstream ENSMUSE00000521490 Chr7:148443931..148446606 GCCGGAACGTGTTACAGAAT Chr7:148444722..148444741 60 50
downstream ENSMUSE00000232784 Chr7:148446859..148447013 AATCCTATGAGGGCCCTGTC Chr7:148446934..148446953 60.29 55
downstream ENSMUSE00000586968 Chr7:148447112..148447441 CATGTTGGGGCTGTAAACCT Chr7:148447207..148447226 59.85 50
downstream ENSMUSE00000513073 Chr7:148447675..148447803 AAGGGTTTAATGGCCAGCTT Chr7:148447742..148447761 59.97 45
downstream ENSMUSE00000393869 Chr7:148448308..148448650 GTGGCTAGCCTCAGGTCTCA Chr7:148448464..148448483 60.56 60
downstream ENSMUSE00000713427 Chr7:148448308..148448494 TCTGCGCATATGCCTGTATT Chr7:148448400..148448419 59.31 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGTCCCAGTACCCAGCTCAT Chr7:148442596..148442616 60.53 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTGATGTGCTTCGTGACTGG Chr7:148442619..148442639 60.46 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000038611