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pBAC-lacZ
Gene/insert name: none
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. 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.