pRS426

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  • Model: PVT4047
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pRS426

PVT4047   2ug

 

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"

                     /note="AmpR promoter"

     rep_origin      4316..5658

                     /note="2u ori"

                     /note="yeast 2u plasmid origin of replication"

     protein_bind    complement(5279..5326)

                     /bound_moiety="FLP recombinase from the Saccharomyces 

                     cerevisiae 2u plasmid"

                     /note="FRT"

                     /note="FLP-mediated recombination occurs in the 8-bp core 

                     sequence TCTAGAAA (Turan and Bode, 2011)."

ORIGIN

        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca

       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg

      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc

      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc

      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg

      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc

      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt

      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat

      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca

      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg

      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg

      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca

      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac

      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa

      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg

      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct

      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac

     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg

     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa

     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa

     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac

     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac

     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat

     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa

     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca

     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg

     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta

     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg

     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa

     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg

     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg

     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg

     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat

     1981 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac ggtatcgata

     2041 agcttgatat cgaattcctg cagcccgggg gatccactag ttctagagcg gccgccaccg

     2101 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca

     2161 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatagga

     2221 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgaggtaact cacattaatt

     2281 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga

     2341 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc

     2401 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg

     2461 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc

     2521 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc

     2581 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga

     2641 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc

     2701 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat

     2761 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg

     2821 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc

     2881 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga

     2941 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact

     3001 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt

     3061 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag

     3121 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg

     3181 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa

     3241 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata

     3301 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg

     3361 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata

     3421 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg

     3481 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct

     3541 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt

     3601 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc

     3661 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga

     3721 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt

     3781 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc

     3841 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa

     3901 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca

     3961 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca

     4021 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct

     4081 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc

     4141 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa

     4201 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt

     4261 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaacga

     4321 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac

     4381 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca

     4441 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt

     4501 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt

     4561 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat

     4621 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc

     4681 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct

     4741 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa

     4801 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga

     4861 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat

     4921 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc

     4981 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta

     5041 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga

     5101 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt

     5161 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg

     5221 cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa agcgctctga

     5281 agttcctata ctttctagag aataggaact tcggaatagg aacttcaaag cgtttccgaa

     5341 aacgagcgct tccgaaaatg caacgcgagc tgcgcacata cagctcactg ttcacgtcgc

     5401 acctatatct gcgtgttgcc tgtatatata tatacatgag aagaacggca tagtgcgtgt

     5461 ttatgcttaa atgcgtactt atatgcgtct atttatgtag gatgaaaggt agtctagtac

     5521 ctcctgtgat attatcccat tccatgcggg gtatcgtatg cttccttcag cactaccctt

     5581 tagctgttct atatgctgcc actcctcaat tggattagtc tcatccttca atgctatcat

     5641 ttcctttgat attggatcat actaagaaac cattattatc atgacattaa cctataaaaa

     5701 taggcgtatc acgaggccct ttcgtc

//

 

Caution:
Product is for research use only!

 

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pRS426,Plasmid pRS426,pRS426 vector

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