Features: insert detection: lacZ' marker(s): ampR promoter: SV40 early replicon: pMB1 terminator: SV40 late enhancer: SV40 early
Applications
contains easily purifiable cassette(s) for construction
reporter construct
vector for other uses
vector permitting visual detection of activity in colonies
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--7.0; HindIII--7.0; EcoRI/HindIII--4.3, 2.7; NotI--3.6 (doublet); BamHI--3.6, 3.2, 0.2.
Reporter plasmid permitting visual detection of beta-galactosidase activity by histochemical staining or fluorometric assay.
Designed to express high levels of beta-galactosidase or an alternative gene product in COS cells. May be used as a reference plasmid in transfection experiments or to monitor the effect of trans-acting factors.
The lacZ gene may be excised by NotI and replaced with alternative sequences.
Recommended ampicillin concentration is 200 ug/ml.
The order of the major features in this plasmid is: pUC19 - EcoRI - SV40 enhancer - SV40 early promoter - SV40 late 16S/19S splice signals - NotI - lacZ - NotI - SV40 late polyadenylation signal - HindIII - pUC19.
Media
ATCC® Medium 1945: Terrific broth (ATCC medium 1743) with 200 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References
MacGregor GR, Caskey CT. Construction of plasmids that express E. coli beta-galactosidase in mammalian cells. Nucleic Acids Res. 17: 2365, 1989. PubMed: 2495524
