| Primary Identifier | MGI:7859517 | Allele Type | Targeted |
| Attribute String | Conditional ready, Inserted expressed sequence, Reporter | Gene | Gt(ROSA)26Sor |
| Transmission | Germline | Strain of Origin | (129S6/SvEvTac x C57BL/6NCrl)F1 |
| Is Recombinase | false | Is Wild Type | false |
| molecularNote | The Gateway Entry system was chosen to guarantee controlled and efficient monosite insertion of the c-MYC construct into the ubiquitously expressed Gt(ROSA)26Sor locus. The toxin-encoding ccdB gene was excised from the pENTR1A vector (Invitrogen) by BamH1/Xho1 restriction digest and a 1.5kb cDNA construct of human c-MYC was inserted into these sites of the pENTR1A vector. BamH1/Xho1 restriction digest showed correct insertion of the construct. Correct orientation of the insert was confirmed by DNA sequencing. The c-MYC construct was subsequently inserted into a targeting vector via in vitro recombination. Since the c-MYC construct in the pENTR1A vector is flanked by specific lambda phage integrase recognition sites (attL), the c-MYC construct could be efficiently transferred to a targeting vector carrying the corresponding heterotypic sites (attR). The targeting vector contains a 5 homology region to the mouse Gt(ROSA)26Sor genomic locus, a splice acceptor (SA) site, a PGK-neo-3x pA stop cassette flanked by loxP-sites (LSL), the c-MYC construct, an IRES-eGFP reporter gene, a 3 homology region to the mouse ROSA26 genomic sequence and a Diphteria Toxin A (DTA) selection cassette. |
