pRS426 Plasmid

$216.00

  • Model: PVT4047
  • 50 Units in Stock
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pRS426

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pRS426,Plasmid pRS426,pRS426 vector

 

pRS426 Information

Promoter: URA3, T7, T3

Replicator: 2 ori, ori, FI ori

Plasmid classification: yeast series, Saccharomyces cerevisiae expression vector

Plasmid size: 5726bp

Prokaryotic resistance: Amp

Screening markers: URA3

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

 

pRS426 Desciption

The order of the major features in this plasmid is:ori(f1) - lacZ - T7 promoter - MCS (KpnI-SacI) - T3 promoter - lacI - ori(pMB1) - ampR - ori (2 micron) - URA3. 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 non-selective growth, plasmids are lost through mitotic segregation at rates in the range of 4.4 + or - 1.4% of progeny per doubling. YE-type shuttle vector encoding the beta-galactosidase alpha peptide, permitting blue-white visual detection. Can be used to produce ssDNA. Contains promoters for in vitro RNA synthesis and priming sites useful for sequencing. Restriction digests of the clone give the following sizes (kb): BamHI--5.8; EcoRI--5.8; PstI--4.0, 1.7; PvuII--5.2, 0.54. (ATCC staff) Medium is 1227 LB plus ampicillin. NCBI gi: 416322 Hosts: E.coli DH5alpha, Saccharomyces cerevisiae, E.coli.

pRS426-vector

 

pRS426 Sequence

LOCUS       Exported                5726 bp ds-DNA     circular SYN 12-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 8
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, September 12, 2016 from SnapGene Viewer 3.1.4
             
FEATURES             Location/Qualifiers
     source          1..5726
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        201..416
                     /gene="S. cerevisiae URA3"
                     /note="URA3 promoter"
     CDS             417..1220
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /note="URA3"
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1589..2167)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     1951..1967
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1977..1995
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2004..2111
                     /note="MCS"
                     /note="pBluescript multiple cloning site"
     primer_bind     2021..2037
                     /note="KS primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2071..2087)
                     /note="SK primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2124..2142)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2163..2179)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2187..2203
                     /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(2211..2241)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(2565..3153)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3324..4184)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4185..4289)
                     /gene="bla"
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