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URL: http://www.addgene.org/16485
Proper Citation: RRID:Addgene_16485
Bacterial Resistance: Ampicillin
Defining Citation: PMID:8755583
Vector Backbone Description: Backbone Size:0; Vector Backbone:pSAR-MT; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: The pSAR-MT inducible expression plasmid was constructed as follows. Two oligonucleotides (5'-GATCTCGAGCTCCCTGCA-3' and 5'-GGGAGCTCGA-3') were annealed, and the resulting fragment was cloned into the BamHI and PstI sites of the scaffold attachment region (SAR)-containing plasmid p8. The resulting plasmid, pJM7, consisted of two SAR sequences flanking a short polylinker containing restriction sites for Xhol and PstI. The mutant metallothionein (MT) promoter was amplified by PCR and cloned into pCEP4, linearized with BglII and NotI, yielding pCEP-MT. A SalI-BamHI fragment containing an intron from the globin gene was then inserted into the XhoI and BamHI sites of pCEP-MT, yielding pCEP-MTi. Finally, the MT promoter/intron/poly(A) region was removed from pCEP-MTi by digestion with Sall and cloned into the XhoI site of pJM7, yielding pSAR-MT.
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Source: Addgene
