YEP352

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  • Model: PVT4038
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YEP352

PVT4038           2ug
 

 

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 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt

     2041 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa

     2101 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa

     2161 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg

     2221 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag

     2281 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg

     2341 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc

     2401 ataacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag

     2461 gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc

     2521 attttcggaa gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc

     2581 tctagctaga aagtatagga acttcagagc gcttttgaaa accaaaagcg ctctgaagac

     2641 gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata

     2701 ccgcttccac aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc

     2761 tatataacct acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca

     2821 ttttttatgt ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc

     2881 atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga

     2941 caaaatagaa gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact

     3001 ttctgttcac aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta

     3061 tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag

     3121 gcttttttta tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc

     3181 tacagtgcaa aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta

     3241 atctaagatg ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa

     3301 gaagtataga ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa

     3361 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca

     3421 aaaatgaagc acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg

     3481 taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt

     3541 ctacaaaatg aagcacagat gcttcgttaa caaagatatg ctattgaagt gcaagatgga

     3601 aacgcagaaa atgaaccggg gatgcgacgt gcaagattac ctatgcaata gatgcaatag

     3661 tttctccagg aaccgaaata catacattgt cttccgtaaa gcgctagact atatattatt

     3721 atacaggttc aaatatacta tctgtttcag ggaaaactcc caggttcgga tgttcaaaat

     3781 tcaatgatgg gtaacaagag cttttcaatt catcattttt tttttattct tttttttgat

     3841 ttcggtttct ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga

     3901 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt

     3961 gcccagtatt cttaacccaa ctgcacagaa caaaaaccga aacgaagata aatcatgtcg

     4021 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt

     4081 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag

     4141 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg

     4201 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc

     4261 aagtacaatt ttttactctt cgaagacaga aaatttgctg acattggtaa tacagtcaaa

     4321 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac

     4381 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag

     4441 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga

     4501 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt

     4561 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc

     4621 ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat

     4681 gtggtctcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg

     4741 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga

     4801 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa

     4861 attagagctt caatttaatt atatcagtta ttaccctatg cggtgtgaaa taccgcacag

     4921 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg

     4981 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaagg ggggatgtgc

     5041 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac

     5101 ggccagtgcc aagcttgcat gcctgcaggt cgactctaga ggatccccgg gtaccgagct

     5161 cgaattcgta atcatggtca t

//

 

Caution:
Product is for research use only!

 

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