Primary Identifier | MGI:5303336 | 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 Ple194-Cre/EGFP plasmid (pEMS1070) was created by inserting the Ple194 MiniPromoter (derived from specific promoter/enhancer/regulatory regions of the human solute carrier family 6 (neurotransmitter transporter, dopamine), member 3 (SLC6A3) gene) into the multiple cloning site of the pEMS1301 plasmid vector backbone. This placed the Ple194 MiniPromoter upstream of a minimal wildtype frt site, a Cre/EGFP fusion gene (Cre recombinase gene and enhanced green fluorescent protein (EGFP; with mutated TAA toTTA 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 Ple194-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. |