pHIS2 Plasmid


  • Model: PVT4037
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pHIS2,Plasmid pHIS2,pHIS2 vector

pHIS2 Plasmid information

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 characterizeDNA-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 andused as baits to screen GAL4 AD/cDNA fusion libraries for proteins that interact with the targetsequence. 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 driveexpression of the HIS3 reporter gene, which enables the host cell to grow on histidine-deficientmedia.

pHIS2 Plasmid Sequence

LOCUS       Exported                7207 bp ds-DNA     circular SYN 23-二月-2016
DEFINITION  synthetic circular DNA
KEYWORDS    Untitled 4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7207)
  TITLE     Direct Submission
  JOURNAL   Exported 2016-2-23 from SnapGene Viewer 3.0.3
FEATURES             Location/Qualifiers
     source          1..7207
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             152..814
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /note="yeast auxotrophic marker"
     promoter        2576..2857
                     /gene="S. cerevisiae TRP1"
                     /note="TRP1 promoter"
     CDS             2858..3532
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /note="yeast auxotrophic marker"
     promoter        complement(3614..3632)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3653..3669)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
     protein_bind    3677..3693
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3701..3731)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3746..3767
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4055..4643)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
     CDS             complement(4814..5608)
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="confers resistance to neomycin, kanamycin, and G418 
     promoter        complement(5609..5713)
                     /note="AmpR promoter"
     misc_feature    5746..6247
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     rep_origin      complement(6564..7019)
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     7164..7180
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
     promoter        7187..7205
                     /note="T7 promoter"
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