pSAR-MT
(Plasmid #16485)

• Purpose: (Empty Backbone)
• Depositing Lab: Bert Vogelstein
• Publication: Morin et al Proc Natl Acad Sci U S A. 1996 Jul 23. 93(15):7950-4.

Backbone

• Vector backbone: pSAR-MT
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: None

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: na

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor 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.