Recombineering Website - Plasmids

The following plasmids are currently available:

• PL451 - white tube
• PL452 - blue tube
• PL253 - orange tube
• pIGCN21 - violet tube
• pEL04 - green tube
• pTamp - yellow tube
• pGalK - brown tube

1. PL451: A plasmid containing a neo cassette flanked by two frt sites and one loxP site. The neo gene is expressed both from a prokaryotic promoter (em7) and a eucaryotic promoter (Pgk): Frt-Pgk-em7-Neo-Frt-loxP. (ref)
   • map (PDF)
   • sequence (text)
   • sequence (annotated text)
2. PL452: A plasmid containing a neo cassette flanked by two loxP sites. The neo gene is expressed both from a prokaryotic promoter (em7) and a eucaryotic promoter (Pgk): loxP-Pgk-em7-Neo-loxP. (ref)
   • map (PDF)
   • sequence (text)
   • sequence (annotated text)
3. PL253: A pBluescript-derived plasmid for retrieval of DNA from a BAC. This plasmid contains an Mc1-driven Thymidine Kinase (TK) cassette for negative selection in ES cells. Retrieval of DNA into this vector from a BAC is the first step in the construction of a gene targeting vector. (ref)
   • map (PDF)
   • sequence (text)
   • sequence (annotated text)
4. pIGCN21: A plasmid containing the IRES-EGFPcre-Frt-kan-Frt cassette. This can be used to insert an EGFP-cre fusion gene into a BAC under the control of a gene of interest to make a tissue-specific Cre mouse. (ref)
   • map (PDF)
   • sequence (text)
   • sequence (annotated text)
5. pEL04: A plasmid containing the Cam-SacB cassette to be used for positive/negative selection. Positive selection using chloamphenicol, and negative selection against the SacB gene using sucrose. (ref)
   • map (PDF)
   • sequence (text)
   • sequence (annotated text)
6. pTamp: A plasmid containing an amp cassette for replacing the loxP site in pBeloBAC11. (ref)
   • map (PDF)
   • sequence (text)
   • sequence (annotated text)
7. pGalK: A galK expression vector for use together with either SW102, SW105, or SW106 in galK positive/negative selection-based BAC recombineering. galK is expressed from the procaryotic em7 promoter. Homology arms can be added using PCR or cloned into this cassette using the unique flanking restriction sites. ( ref )
   • map (PDF)
   • sequence (text)
   • sequence (annotated pdf)