|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||13422||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 11151
Vector typeBacterial Expression, Cre/Lox
Growth in Bacteria
Copy numberLow Copy
- 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 (Common Sequencing Primers)
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pBAC-lacZ was a gift from Keith Joung (Addgene plasmid # 13422 ; http://n2t.net/addgene:13422 ; RRID:Addgene_13422)