Plasmid 13423: pBAC-BA-lacZ

• Gene/insert name: BA
• Alt name: Binding site for original BCR-ABL three-finger array
• Vector backbone: pBAC-lacZ
  (Search Vector Database)
• Vector type: Bacterial Expression, Cre/Lox
• Backbone size w/o insert (bp): 11151
• Cloning site 5': BsaI
• Site destroyed during cloning: No
• Cloning site 3': BsaI
• Site destroyed during cloning: No
• 5' sequencing primer: CGC CAG GGT TTT CCC AGT CAC GAC
• Bacterial resistance(s) Chloramphenicol
• Growth strain(s) DH5alpha
• Growth temperature (℃): 37
• High or low copy: High Copy
• Sequence: View sequences (2)
• Map: View map
• Principal Investigator: Keith Joung
• Terms and Licenses: MTA

Comments: 

The complete assembled sequence for this plasmid may not be accurate. Addgene suggests sequence verifying the construct before starting your experiments.

Bacterial cell-based two-hybrid (B2H) reporter vector with binding site for "original" BCR-ABL three-finger array for use as a positive control with pGP-FB-orig BA. Use 12.5 ug/ml chloramphenicol for growth.

The pBAC-lacZ plasmid is a mini-F' that can be replicated in
standard E. coli strains, but because it is maintained as a single copy episome, it
gives low DNA yields. pBAC-lacZ also contains a second, higher copy number
origin of replication (oriV) that is only active in the presence of a trans-acting
factor encoded by the trfA gene. Transformax EPI300 cells express trfA from an
inducible promoter that we have found is inducible with arabinose. When the trfA gene is induced in Transformax EPI300 cells, the copy number of pBAC-lacZ plasmid is increased in the cells and reasonable yields of plasmid can be obtained using a standard miniprep procedure.

Further notes on propagating this plasmid: grow an overnight culture (without arabinose)
and then the next morning subculture 1:10 into medium that has a final
concentration of arabinose of 1 mM. You then grow this culture for 5 to 6 hours
at 37 C and then harvest the DNA.