Addgene - pBAC-lacZ Plasmid Data

Plasmid 13422: pBAC-lacZ

Gene/insert name:
None
Vector backbone:
pBAC-lacZ
(Search Vector Database)
Vector type:
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
Bacterial resistance(s)
Chloramphenicol
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
Low Copy
Sequence: View sequences (2)
Map:
View map
Supplemental document:
Notes from Addgene (1)
Principal Investigator:
Keith Joung
Terms and Licenses:
MTA

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.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 13422" in your Materials and Methods section.