pRS403 Plasmid

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  • Model: PVT4043
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pRS403 Plasmid

PVT4043     2ug

 

pRS403 Plasmid  Information

Copier: pUC ori, FIori

Plasmid Classification: Yeast Series Plasmids; Yeast Expression Plasmids; Saccharomyces cerevisiae Plasmids

Plasmid size: 4456 BP

Prokaryotic resistance: ampicillin Amp (100 ug/ml)

Screening Marker: HIS3

Cloning strains: Escherichia coli such as DH5alpha

Culture conditions: 37 ℃, aerobic LB

Expression host: Saccharomyces cerevisiae YPH499, YPH500, etc.

Culture conditions: 30℃, YPD, aerobic

5'Sequencing Primer: T7 (TAATACGACTCACTATAGG)

3'Sequencing Primers: M13R (CAGGAAACAGCTATGACC)

 

pRS403 Plasmid Description

PRS4xx series plasmids were used to clone Saccharomyces cerevisiae by shuttle vector of gene, but they could replicate in E. coli. Although in E. coli, they replicated from the origin of pMB1 to pBR322. The bla (APR) markers they carried were used to select single-stranded DNA from the origin of bacteriophage F1 using ampicillin and replication.pRS403 through pRS406 are integrative plasmids (YIp); pRS413through pRS416 (shown below) are centromeric plasmids (YCp),and differ from the YIp by the insertion of a centromere (CEN)and autonomously replicating sequence (ARS); pRS423 throughpRS426 are episomal plasmids (YEp), and carry instead theorigin of replication from the yeast 2µ circle plasmid along withREP3 and FRT sequences (2). The 4 vectors in each series differonly in the yeast auxotrophic marker used: HIS3 (pRS4x3), TRP1(pRS4x4), LEU2 (pRS4x5) or URA3 (pRS4x6).

pRS403-plasmid

 

Publication for pRS403 Plasmid PVT4043

Structural basis for the interaction of the chaperone Cbp3 with newly synthesized cytochrome b during mitochondrial respiratory chain assembly
Mama Ndi1, Geoffrey Masuyer1, Hannah Dawitz1, Andreas Carlström1, Mirco Michel1, Arne Elofsson2, Mikaela Rapp1, Pål Stenmark3 and Martin Ott3*

First Published on Journal of biological chemistry
September 19, 2019
doi: 10.1074/jbc.RA119.010483
jbc.RA119.010483.

 

pRS403 Plasmid Sequence

LOCUS       Exported                4456 bp ds-DNA     circular SYN 29-NOV-2013

DEFINITION  Yeast integrative vector with a HIS3 marker and an MCS derived from 

            pBLUESCRIPT II.

ACCESSION   U03443

KEYWORDS    pRS403

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 4456)

  AUTHORS   Sikorski RS, Hieter P.

  TITLE     A system of shuttle vectors and yeast host strains designed for 

            efficient manipulation of DNA in Saccharomyces cerevisiae.

  JOURNAL   Genetics 1989;122:19-27.

  PUBMED    2659436

REFERENCE   2  (bases 1 to 4456)

  TITLE     Direct Submission

  JOURNAL   Exported Monday, September 12, 2016

FEATURES             Location/Qualifiers

     source          1..4456

                     /organism="synthetic DNA construct"

                     /lab_host="Saccharomyces cerevisiae"

                     /mol_type="other DNA"

     promoter        317..503

                     /gene="S. cerevisiae HIS3"

                     /note="HIS3 promoter"

     CDS             504..1163

                     /codon_start=1

                     /gene="S. cerevisiae HIS3"

                     /product="imidazoleglycerol-phosphate dehydratase, required

                     for histidine biosynthesis"

                     /note="HIS3"

                     /note="yeast auxotrophic marker"

                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE

                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF

                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL

                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"

     rep_origin      complement(1423..1878)

                     /direction=LEFT

                     /note="f1 ori"

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     CDS             complement(1661..2239)

                     /codon_start=1

                     /gene="lacZ fragment"

                     /product="LacZ-alpha fragment of beta-galactosidase"

                     /note="lacZ-alpha"

                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI

                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR

                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA

                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"

     primer_bind     2023..2039

                     /note="M13 fwd"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     promoter        2049..2067

                     /note="T7 promoter"

                     /note="promoter for bacteriophage T7 RNA polymerase"

     misc_feature    2076..2183

                     /note="MCS"

                     /note="pBluescript multiple cloning site"

     promoter        complement(2196..2214)

                     /note="T3 promoter"

                     /note="promoter for bacteriophage T3 RNA polymerase"

     primer_bind     complement(2235..2251)

                     /note="M13 rev"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     protein_bind    2259..2275

                     /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(2283..2313)

                     /note="lac promoter"

                     /note="promoter for the E. coli lac operon"

     rep_origin      complement(2637..3225)

                     /direction=LEFT

                     /note="ori"

                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 

                     replication"

     CDS             complement(3396..4256)

                     /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(4257..4361)

                     /gene="bla"

                     /note="AmpR promoter"

ORIGIN

        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca

       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg

      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc

      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt

      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta

      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat

      361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata

      421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc

      481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa

      541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact

      601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga

      661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt

      721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca

      781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag

      841 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag

      901 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag

      961 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta

     1021 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca

     1081 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct

     1141 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat

     1201 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat

     1261 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt

     1321 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt

     1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata

     1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg

     1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc

     1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa

     1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt

     1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac

     1741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta

     1801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg

     1861 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc

     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc

     1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg

     2041 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg

     2101 acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact agttctagag

     2161 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc

     2221 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca

     2281 cacaacatag gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgaggtaa

     2341 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag

     2401 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc

     2461 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct

     2521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg

     2581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc

     2641 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga

     2701 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct

     2761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg

     2821 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag

     2881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat

     2941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac

     3001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac

     3061 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc

     3121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt

     3181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc

     3241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg

     3301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca

     3361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca

     3421 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag

     3481 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac

     3541 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc

     3601 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct

     3661 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc

     3721 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg

     3781 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc

     3841 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat

     3901 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag

     3961 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat

     4021 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg

     4081 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca

     4141 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga

     4201 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc

     4261 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata

     4321 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg

     4381 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc

     4441 acgaggccct ttcgtc

//

 

Caution:
Product is for research use only!

 

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pRS403,Plasmid pRS403,pRS403 vector

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