pBAC-BA-lacZ
(Plasmid #13423)

• Depositing Lab: Keith Joung
• Publication: Wright et al Nat Protoc. 2006;1(3):1637-52. doi: 10.1038/nprot.2006.259

Backbone

• Vector backbone: pBAC-lacZ
• Backbone size w/o insert (bp): 11151
• Vector type: Bacterial Expression, Cre/Lox

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: BA
• Alt name: Binding site for original BCR-ABL three-finger array

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (not destroyed)
• 3′ cloning site: BsaI (not destroyed)
• 5′ sequencing primer: CGC CAG GGT TTT CCC AGT CAC GAC

Resource Information

• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor 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.