PHIS2 plasmid - 2 ug

Promoters: TRP1, T7, T3

Replicon: pUC ori, FI ori

Plasmid classification: yeast series, yeast one hybrid vector

Plasmid size: 7207bp

Prokaryotic resistance: Kan

Selection marker: TRP1

Clone strain: DH5 alpha

Culture conditions: 37 LB, aerobic

Host cells: yeast cells

Primers for 5'sequencing: M13F:TGTAAAACGACGGCCAGT

Primers for 3'sequencing: primers were designed according to the sequence

Use: Yeast expression

pHIS2 Plasmid description

pHIS2 is a reporter vector that can be used in yeast one-hybrid assays to identify and characterize DNA-binding proteins. The vector was specifically designed for use with the BD Matchmaker™ One-Hybrid Library Construction & Screening Kit (#K1617-1). It contains a HIS3 nutritional reporter gene, located downstream of a multiple cloning site (MCS) and the minimal promoter of the HIS3 locus(PminHIS3). Cis-acting DNA sequences, or DNA target elements, can be inserted into the MCS and used as baits to screen GAL4 AD/cDNA fusion libraries for proteins that interact with the target sequence. A protein-DNA (or one-hybrid) interaction can be detected by performing the assay in ayeast strain such as Y187 that is auxotrophic for histidine. Positive one-hybrid interactions drive expression of the HIS3 reporter gene, which enables the host cell to grow on histidine-deficient media.