Plasmids

The following plasmids are currently available:

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.
Reference
map (PDF)
sequence (text)
sequence (annotated text)

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.
Reference
map (PDF)
sequence (text)
sequence (annotated text)

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.
Reference
map (PDF)
sequence (text)
sequence (annotated text)

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.
Reference
map (PDF)
sequence (text)
sequence (annotated text)

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.
Reference
map (PDF)
sequence (text)
sequence (annotated text)

pTamp: A plasmid containing an amp cassette for replacing the loxP site in pBeloBAC11.
Reference
map (PDF)
sequence (text)
sequence (annotated text)

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.
Reference
map (PDF)
sequence (text)
sequence (annotated pdf)