| Primary Identifier | MGI:5303289 | Allele Type | Targeted |
| Attribute String | Recombinase, Reporter | Gene | Hprt1 |
| Transmission | Cell Line | Strain of Origin | 129P2/OlaHsd |
| Is Recombinase | true | Is Wild Type | false |
| Project Collection | Pleiades Promoter Project |
| molecularNote | The promoterless-Cre/EGFP plasmid (pEMS1301) was created by inserting no Pleiades Promoter into the multiple cloning site of the pEMS1301 plasmid vector backbone. Therefore, no promoter is upstream of a minimal wildtype frt site, a Cre/EGFP fusion gene (Cre recombinase gene and enhanced green fluorescent protein (EGFP; with mutated TAA->TTA stop)), a nuclear localization signal, an SV40 early polyA signal, a second minimal wildtype frt site , a human HPRT complementary sequence (containing exon 1, intron 1, exon 2, and part of intron 2), a mouse 3' Hprt homology arm, an I-Sce linearization cut site, and a mouse 5' Hprt homology arm. Of note, the sequences between the two frt sites are in reverse orientation compared to the HPRT sequences. This promoterless-Cre/EGFP construct was targeted as a single copy knock-in to the Hprtb-m3 mutant locus on the X Chromosome via electroporation into E14TG2a embryonic stem cells: a male ES cell line harboring the Hprtb-m3 mutation on the X Chromosome. The resulting ES cells, correctly targeted with the promoterless-Cre/EGFP transgene inserted into the Hprt locus, were assigned Hprttm205(cre/EGFP)Ems. |
