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URL: http://www.addgene.org/13422

Proper Citation: RRID:Addgene_13422

Bacterial Resistance: Chloramphenicol

Defining Citation: PMID:17406455

Vector Backbone Description: Backbone Size:11151; Vector Backbone:pBAC-lacZ; Vector Types:Bacterial Expression, Cre/Lox; Bacterial Resistance:Chloramphenicol

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. 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.

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