Primary Identifier | MGI:5806633 | Allele Type | Targeted |
Attribute String | Recombinase | Gene | Gnb4 |
Transmission | Germline | Strain of Origin | (129S6/SvEvTac x C57BL/6NCrl)F1 |
Is Recombinase | true | Is Wild Type | false |
molecularNote | The targeting vector contained, from 5' to 3', a partial Gnb4 sequence spanning intron 6 through intron 9, an frt3 site within Gnb4 intron 9, a partial Gnb4 exon 10 sequence up to and including the endogenous stop codon, an internal ribosome entry site 2 (IRES2) sequence (allows translation initiation in the middle of an mRNA sequence), a Cre recombinase gene, a bovine growth hormone polyA sequence, an AttB site, a PGK promoter-Neomycin resistance gene-PGK polyA cassette, an frt5 site, an mRNA splice acceptor sequence, the 3' portion of the hygromycin gene (Hygro2) with SV40 polyA signal, and an AttP site. This construct was electroporated into (129S6/SvEvTac x C57BL/6)F1-derived G4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. Chimeric mice were bred to PhiC31-expressing mice to remove the the AttB/AttP-flanked PGK-neo sequence and replace it with the recombined AttB/AttP site (AttL). |