pRS425 Plasmid


  • Model: PVT4046
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pRS425,Plasmid pRS425,pRS425 vector

pRS425 plasmid informaiton

Promoter: LEU2, T7, T3
Replicator: 2 ori, ori, FI ori
Plasmid classification: yeast series, Saccharomyces cerevisiae expression vector
Plasmid size: 6849bp
Prokaryotic resistance: Amp
Screening markers: LEU2
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic LB
Expression host: yeast cells
Induction method: galactose induction
5'sequencing primers: M13R:CAGGAAACAGCTATGACC
3'sequencing primers: primers designed according to sequence

pRS425 plasmid Description
This is a YEtype(episomal) yeast shuttle vector. It encodes the beta galactosidase alpha peptide for bluewhite color detection of inserts and also has the T7 and T3 promoters for in vitro RNA synthesis and priming sites for sequencing. It also contains the REP3 and FRT sequences necessary for high copy propagation in yeast. In S. cerevisiae, the copy number is about 20 per haploid cell. In nonselective growth, plasmids are lost through mitotic segregation at rates in the range of 4.6 +/ 0.6% of progeny per doubling. The order of the major features in this plasmid is: ori(f1) lacZ T7 promoter MCS (KpnISacI) T3 promoter lacI ori(pMB1) ampR ori(2 micron) LEU2.

pRS425 plasmid Sequence

LOCUS       Exported                6849 bp ds-DNA     circular SYN 12-SEP-2016
DEFINITION  synthetic circular DNA
KEYWORDS    Untitled 7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6849)
  TITLE     Direct Submission
  JOURNAL   Exported Monday, September 12, 2016 from SnapGene Viewer 3.1.4
FEATURES             Location/Qualifiers
     source          1..6849
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(663..1757)
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /note="yeast auxotrophic marker"
     promoter        complement(1770..2174)
                     /gene="S. cerevisiae LEU2"
                     /note="LEU2 promoter"
     rep_origin      complement(2474..2929)
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(2712..3290)
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
     primer_bind     3074..3090
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
     promoter        3100..3118
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3127..3234
                     /note="pBluescript multiple cloning site"
     primer_bind     3144..3160
                     /note="KS primer"
                     /note="common sequencing primer, one of multiple similar 
     primer_bind     complement(3194..3210)
                     /note="SK primer"
                     /note="common sequencing primer, one of multiple similar 
     promoter        complement(3247..3265)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3286..3302)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
     protein_bind    3310..3326
                     /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(3334..3364)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(3688..4276)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
     CDS             complement(4447..5307)
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
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