pKJ1267 (aka pAC-alphaGal4)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||21869||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size (bp) 3428
Vector typeBacterial Expression
Growth in Bacteria
Growth Strain(s)XL1 Blue
Growth instructionsGrow 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).
Copy numberLow Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer lac promoter (Common Sequencing Primers)
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)
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:pKJ1267 (aka pAC-alphaGal4) was a gift from Keith Joung (Addgene plasmid # 21869 ; http://n2t.net/addgene:21869 ; RRID:Addgene_21869)