Addgene - pKJ1267 (aka pAC-alphaGal4) Plasmid Data

Plasmid 21869: pKJ1267 (aka pAC-alphaGal4)

Gene/insert name:
None
Vector backbone:
pKJ1267
(Search Vector Database)
Vector type:
Bacterial Expression
Backbone size (bp):
3428
5' sequencing primer:
lac promoter List of Sequencing Primers
Bacterial resistance(s)
Chloramphenicol
Growth strain(s)
XL1 Blue
Growth temperature (℃):
37
Growth instructions:
Grow in LB supplemented with 30 µg/ml chloramphenicol. Due to toxicity associated with high expression from the dual lpp/lacUV5 promoter, this plasmid must be propagated in a lacIq strain (e.g.--XL-1 Blue).
High or low copy:
Low Copy
Sequence: View sequences (2)
Supplemental document:
Notes from Addgene (1)
Principal Investigator:
Keith Joung
Terms and Licenses:
MTA

Comments:

Bacterial expression vector for the alpha-Gal4 hybrid protein (required for the B2H system). The tandem lpp/lacUV5 promoter which drives expression of the Gal11P fusion protein contains a lac operator and therefore these dual promoters can be repressed by lac repressor.

This plasmid is a very low copy number plasmid and therefore we recommend that maxipreps be performed from a 200 ml culture. A miniprep will not be sufficient.

Annotations for pKJ1267:

2988- 219: Chloramphenicol-resistance gene (complementary strand)/ 581-1493: p15A origin of replication/ 1552-1557: lpp promoter -35 box / 1576-1581: lpp promoter -10 box / 1624-1629 : lacUV5 promoter -35 box / 1648-1653 : lacUV5 promoter -10 box / 1748-2623 : alpha-Gal4 hybrid protein gene (E. coli RNA polymerase alpha subunit aa 1-248 fused by an Ala-Ala-Ala linker to aa 58-97 of the yeast Gal4 protein)

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 21869" in your Materials and Methods section.