YEP352 Plasmid

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  • Model: PVT4038
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YEP352

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YEP352,Plasmid YEP352,YEP352 vector

 

YEP352 Informaiton

Promoter: URA3
Replicator: 2 ori, ori
Plasmid classification: yeast series, Saccharomyces cerevisiae expression vector
Plasmid size: 5881bp
Prokaryotic resistance: Amp
Screening markers: URA3
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic LB
Expression host: yeast cells
5'sequencing primers: M13F:TGTAAAACGACGGCCAGT
3'sequencing primers: primers designed according to sequence
Use: Yeast expression

 

YEP352 Description

Restriction digests of the clone give the following sizes (kb): BamHI--5.3; XbaI--5.3; PstI--5.3; HindIII--5.3.This is a Escherichia coli/yeast episomal shuttle vector with multiple restriction sites for cloning.Constructed by filling in and blunt-end ligating pUC18 linearized at NdeI, a 1.4 kb RsaI fragment of the 2 micron circle, and a 1.1 kb HindIII/SmaI fragment containing URA3. A XbaI site in 2u and a PstI site in URA3 were removed.The yeast genomicsequence is in nt 286-1383 (where the A of the ATG of lacZ' is number 1), the 2 micron sequence goes from nt 1384-2768, and pUC18 includes nt 2769-285.

YEP352-plasmid

 

YEP352 Sequence

LOCUS       Exported                5181 bp ds-DNA     circular SYN 11-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5181)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, September 11, 2016 from SnapGene Viewer 3.1.4
            
FEATURES             Location/Qualifiers
     source          1..5181
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    20..36
                     /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(44..74)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(398..986)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1157..2017)
                     /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(2018..2122)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      2404..3568
                     /note="2u ori"
                     /note="yeast 2u plasmid origin of replication"
     promoter        3804..4014
                     /gene="S. cerevisiae URA3"
                     /note="URA3 promoter"
     CDS             4015..4818
                     /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"
     CDS             complement(4891..5181)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIG"
     primer_bind     5091..5107
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5111..5167
                     /note="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5177..12)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
       61 gcataaagtg taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc
      121 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctggattaa tgaatcggcc
      181 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
      241 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      301 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      361 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      421 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      481 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      541 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
      601 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      661 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      721 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      781 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
      841 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      901 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      961 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1021 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1081 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1141 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1201 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1261 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1321 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1381 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1441 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1501 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1561 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1621 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1681 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1741 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1801 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1861 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1921 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1981 agggaataa
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