pSC65
(Plasmid #30327)

• Depositing Lab: Bernard Moss
• Publication: Chakrabarti et al Biotechniques. 1997 Dec . 23(6):1094-7.

Backbone

• Vector backbone: pBR322
• Backbone manufacturer: n/a
• Backbone size w/o insert (bp): 4500
• Total vector size (bp): 7253
• Modifications to backbone: Addition of vaccinia virus TK sequence flanking the lacZ gene, dual synthetic early-late promotor, 7.5 promotor and multiple cloning site with various restriction enzyme sites.
• Vector type: Vaccinia virus expression vector

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Growth instructions: PROTOCOL FOR AMPLIFICATION OF TRANSFORMED LOW COPY NUMBER PLASMIDS IN ECOLI, (i.e. pSC 63, pSC 59, pSC 65 and pOS-7) After transformation, grow Ecoli (10-12 hrs) in 2XY-T media (LB is acceptable but plasmid yield will be lower) to log phase containing 100-150 mcg/ml ampicillin or carbenicillin (if stock is 1000X-(i.e.100-150 mg/ul), use 1 ul per 1 ml media) final concentration. At this point add chloramphenicol(dissolved in ethanol) to a final concentration of 25 mcg/ml (if stock is 1000x-(i.e.25 mg/ml) use 1 ml per ml media). Continue growing an additional 12-14 hrs. before harvesting cells.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: None
• Promoter: synthetic early-late; 7.5 promotors

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: n/a (unknown if destroyed)
• 3′ cloning site: n/a (destroyed during cloning)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: n/a