Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI31181
Trapped Gene
4930434E21Rik (ENSMUSG00000029658)
Vector Insertion
Chr 5: 150355064 - 150365790
Public Clones IST12678C1 (tigm) IST12384H4 (tigm) IST12438B6 (tigm) IST11110E4 (tigm)
IST12101H2 (tigm) IST12946E1 (tigm) IST12696C10 (tigm) IST12722F5 (tigm)
IST12101H2 (tigm) IST12644B10 (tigm) IST12757G9 (tigm) IST11110E4 (tigm)
IST12485G11 (tigm) IST12693F1 (tigm) IST12310H3 (tigm) IST11785G5 (tigm)
IST14959E8 (tigm) IST12102H2 (tigm) IST12987E2 (tigm) IST12397B2 (tigm)
IST12791D11 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 11% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000718744 (Chr5:150354994..150355063 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ATTTCCAGCTCCAATCACGA Chr5:150355024..150355043 60.6 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000718744 (Chr5:150354994..150355063 +)
Downstram Exon
ENSMUSE00000721169 (Chr5:150365791..150365820 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ATTTCCAGCTCCAATCACGA Chr5:150355024..150355043 60.6 45 TCCAAGCGGTTTAGGAGAGA Chr5:150365815..150365834 59.95 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000684183 Chr5:150331254..150331346 ACAGGCTGATGCCTCTTTGT Chr5:150331298..150331317 59.87 50
upstream ENSMUSE00000589263 Chr5:150332266..150332380 TGGCACCGCTAAAGTAGTCA Chr5:150332301..150332320 59.49 50
upstream ENSMUSE00000712758 Chr5:150332308..150332380 GGACTGGATCGAGGGAAGAG Chr5:150332323..150332342 61.13 60
upstream ENSMUSE00000533218 Chr5:150335235..150335281 TGTTTTTGGCATCTGTAGGTGA Chr5:150335251..150335272 60.53 40.91
upstream ENSMUSE00000721302 Chr5:150335235..150335281 TGTTTTTGGCATCTGTAGGTGA Chr5:150335251..150335272 60.53 40.91
upstream ENSMUSE00000533217 Chr5:150347317..150347434 AGTATGTCCGCTGGAAGGTG Chr5:150347394..150347413 60.13 55
upstream ENSMUSE00000713315 Chr5:150347317..150347434 AGTATGTCCGCTGGAAGGTG Chr5:150347394..150347413 60.13 55
upstream ENSMUSE00000533216 Chr5:150354994..150355063 ATTTCCAGCTCCAATCACGA Chr5:150355024..150355043 60.6 45
upstream ENSMUSE00000718744 Chr5:150354994..150355063 ATTTCCAGCTCCAATCACGA Chr5:150355024..150355043 60.6 45

*** Putative Vector Insertion (Chr 5: 150355064 - 150365790) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000647562 Chr5:150365791..150365883 TCCAAGCGGTTTAGGAGAGA Chr5:150365815..150365834 59.95 50
downstream ENSMUSE00000709148 Chr5:150365791..150365883 TCCAAGCGGTTTAGGAGAGA Chr5:150365815..150365834 59.95 50
downstream ENSMUSE00000721169 Chr5:150365791..150365820 TCCAAGCGGTTTAGGAGAGA Chr5:150365815..150365834 59.95 50
downstream ENSMUSE00000589262 Chr5:150366758..150367001 GCTGTTCCCCATCTTGACAT Chr5:150366843..150366862 59.93 50
downstream ENSMUSE00000713379 Chr5:150366758..150367001 GCTGTTCCCCATCTTGACAT Chr5:150366843..150366862 59.93 50
downstream ENSMUSE00000533211 Chr5:150366933..150367001 CCAGACCCTGATGATTCGAT Chr5:150366987..150367006 59.89 50
downstream ENSMUSE00000533210 Chr5:150368938..150369043 No primer for this exon
downstream ENSMUSE00000259942 Chr5:150376564..150376715 CCCACTTTCAGGTGAAGGAA Chr5:150376718..150376737 60.08 50
downstream ENSMUSE00000533206 Chr5:150376564..150376715 CCCACTTTCAGGTGAAGGAA Chr5:150376718..150376737 60.08 50
downstream ENSMUSE00000259938 Chr5:150381629..150381731 ACGTACCAGGTGGGACTCTG Chr5:150381659..150381678 60.03 60
downstream ENSMUSE00000533204 Chr5:150381629..150381731 ACGTACCAGGTGGGACTCTG Chr5:150381659..150381678 60.03 60
downstream ENSMUSE00000259929 Chr5:150383191..150383297 GACACCTGTCTTCCCGTTTC Chr5:150383234..150383253 59.56 55
downstream ENSMUSE00000533202 Chr5:150383191..150383297 GACACCTGTCTTCCCGTTTC Chr5:150383234..150383253 59.56 55
downstream ENSMUSE00000444505 Chr5:150383380..150383564 CTGCGAGGCACGATTTTACT Chr5:150383567..150383586 60.41 50
downstream ENSMUSE00000533200 Chr5:150383380..150383459 GGTGTGCAGACAGTGTCCAT Chr5:150383452..150383471 59.58 55
downstream ENSMUSE00000589272 Chr5:150384403..150384457 ATTTCGGTTCACCTCCATGT Chr5:150384458..150384477 59.26 45
downstream ENSMUSE00000589271 Chr5:150385662..150385707 TTATTCTTCGGTCCCATCCA Chr5:150385699..150385718 60.27 45
downstream ENSMUSE00000589270 Chr5:150387032..150387091 GCTTCCGGAAATGATGAAAA Chr5:150387068..150387087 60.02 40
downstream ENSMUSE00000589269 Chr5:150390645..150390810 GTAACTGGAGGTGGCCAGAA Chr5:150390719..150390738 60.11 55
downstream ENSMUSE00000589268 Chr5:150395670..150395765 AACTTCGTCAGCCGTGTCTT Chr5:150395727..150395746 59.91 50
downstream ENSMUSE00000589267 Chr5:150397049..150397107 GAGGCCAGACCCACTAAAGA Chr5:150397092..150397111 59.28 55
downstream ENSMUSE00000589266 Chr5:150397825..150397966 GCCAGGTAAGCGAGAGTGTC Chr5:150397892..150397911 60.02 60
downstream ENSMUSE00000589265 Chr5:150398891..150398930 GAGCTCTCCAGGATGTCACG Chr5:150398914..150398933 60.98 60
downstream ENSMUSE00000589264 Chr5:150400075..150400163 GTGCAGTCAAGGGAGGAAGA Chr5:150400131..150400150 60.39 55
downstream ENSMUSE00000513994 Chr5:150401779..150401972 AACATCCCAGGGGTGATTCT Chr5:150401813..150401832 60.58 50
downstream ENSMUSE00000533168 Chr5:150402207..150402484 TAGGGTTGAGCATGTTGCTG Chr5:150402257..150402276 59.86 50
downstream ENSMUSE00000684162 Chr5:150402727..150402729 No primer for this exon
downstream ENSMUSE00000684164 Chr5:150408864..150408961 TATCTGCTTTCGCCTCTGCT Chr5:150408933..150408952 60.26 50
downstream ENSMUSE00000684163 Chr5:150414071..150414469 AGGCTTTCTTCGAGCTGACA Chr5:150414098..150414117 60.28 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CACGAGCCCACAGCTTTAGT Chr5:150355040..150355060 60.45 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CACGAGCCCACAGCTTTAGT Chr5:150355040..150355060 60.45 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000029658