vector containing primer sites useful for sequencing
vector permitting production of single-stranded DNA
vector permitting visual detection of recombinants
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--5.4; HindIII--5.4; XhoI--5.4.
The AUG start codon is 22bp downstream from the HindIII site within the luc gene.
Differs from pLUCS (ATCC 77415) only in the polylinker.
Permits beta-galactosidase blue/white color selection for identification of recombinants. The color is weaker than from the parent vector and transformed cells may need to be grown 18-24 hours at 37C.
Promoter-cloning vector for analysis of eukaryotic promoters, using the luciferase reporter gene.
Constructed from pSVOALdelta5' by inserting a 2.6 kb BamHI fragment, containing the firefly (Photinus pyralis) luciferase gene and SV40 sequences into the BamHI site of pBluescriptSK+, followed by the removal of the 3' BamHI site.
The order of the major features in the phagemid is: 3' lacZ - XhoI/MCS/BamHI - 5' luc 3' - SV40 small t intron - SV40 polyadenylation signal - NotI - SstII - 5' lacZ - pMB1 ori - ampR - f1 ori.
Media
ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References
de Wet JR, et al. Firefly luciferease gene: Structure and expression in mammalian cells. Mol. Cell. Biol. 7: 725-737, 1987. PubMed: 3821727
Caricasole A, Ward A. A luciferase-reporter vector with blue-white selection for rapid subcloning and mutational analysis of eukaryotic promoters. Gene 124: 139-140, 1993. PubMed: 8382654
Caricasole A, Ward A. Transactivation of mouse insulin-like growth factor II (TGF-II) gene promoters by the AP-1 complex. Nucleic Acids Res. 21: 1873-1879, 1993. PubMed: 8493103
