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    Plasmid map of pBAC-BA-lacZ

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    pBAC-BA-lacZ
    Gene/insert name: BA
    Alternative names: Binding site for original BCR-ABL three-finger array
    Insert size (bp): Unknown
    Vector backbone: pBAC-lacZ (Search Vector Database)
    Type of vector: Bacterial expression,Cre/Lox
    Backbone size (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 (List of Sequencing Primers)
    Bacteria resistance: Chloramphenicol
    High or low copy: High Copy
    Grow in standard E. coli @ 37C: Yes
    Plasmid Provided In: Transformax EPI300
    Principal Investigator: Keith Joung
    Comments: 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.



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