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pESC-ura 构建费用300元/个起

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基本信息

别名:

pESC-URA, pESC URA

出品公司:

Agilent 安捷伦

载体类型:

酿酒酵母蛋白表达载体

载体抗性:

Ampicillin (氨苄青霉素)

载体标签

C-Flag, C-Myc

筛选标记:

His3

启动子

GAL1GAL10

表达水平:

拷贝

载体大小:

6631 bp

克隆方法:

多克隆位点,限制性内切酶

5' 测序引物:

GAL1-F: 5-ATTTTCGGTTTGTATTACTTC-3;

GAL10-F: 5-GGTGGTAATGCCATGTAATATG-3

3' 测序引物

GALI-R: 5-GTTCTTAATACTAACATAACT-3

GAL10-R: 5-GGCAAGGTAGACAAGCCGACAAC-3

稳定性:

稳定 Stable

组成型:

诱导半乳糖

备注

半乳糖诱导,可以同时使两个基因在酿酒酵母中表达。


质粒图谱和多克隆位点信息

注意:

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如要使用本公司保存的母源载体时,本公司不能保证所有母源载体完全符合客户预期,也不承担由此产生的任何责任。


载体描述

带有URA3标记的酵母表达载体,用半乳糖调节表达和标记多达2个基因。

 

载体序列

LOCUS       Exported                6631 bp ds-DNA     circular SYN 03-APR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pESC-URA

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 6631)

  AUTHORS   BT-Lab

  TITLE     Direct Submission

  JOURNAL   Exported Wednesday, Apr 3, 2019 from SnapGene Viewer 4.3.6

FEATURES             Location/Qualifiers

     source          1..6631

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        201..416

                     /gene="S. cerevisiae URA3"

                     /label=URA3 promoter

     CDS             417..1220

                     /codon_start=1

                     /gene="S. cerevisiae URA3"

                     /product="orotidine-5'-phosphate decarboxylase, required

                     for uracil biosynthesis"

                     /label=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

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow

                     indicates direction of (+) strand synthesis"

     terminator      1885..2050

                     /gene="S. cerevisiae ADH1"

                     /label=ADH1 terminator

                     /note="transcription terminator for the S. cerevisiae

                     alcohol dehydrogenase 1 (ADH1) gene"

     CDS             complement(2198..2221)

                     /codon_start=1

                     /product="FLAG(R) epitope tag, followed by an enterokinase

                     cleavage site"

                     /label=FLAG

                     /translation="DYKDDDDK"

     misc_feature    2223..2263

                     /label=MCS1

                     /note="multiple cloning site"

     promoter        2268..2932

                     /gene="GAL1 and GAL10 genes of S. cerevisiae"

                     /label=GAL1,10 promoter

                     /note="divergent inducible promoter, regulated by Gal4"

     protein_bind    2484..2601

                     /label=UAS

                     /bound_moiety="Gal4"

                     /note="upstream activating sequence mediating

                     Gal4-dependent induction"

     promoter        2943..2961

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     misc_feature    2963..2975

                     /label=MCS2

                     /note="multiple cloning site"

     CDS             2979..3008

                     /codon_start=1

                     /product="Myc (human c-Myc proto-oncogene) epitope tag"

                     /label=Myc

                     /translation="EQKLISEEDL"

     terminator      3038..3227

                     /gene="S. cerevisiae CYC1"

                     /label=CYC1 terminator

                     /note="transcription terminator for CYC1"

     rep_origin      complement(3470..4058)

                     /direction=LEFT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of

                     replication"

     CDS             complement(4229..5089)

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     promoter        complement(5090..5194)

                     /gene="bla"

                     /label=AmpR promoter

     rep_origin      5221..6563

                     /label=2u ori

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

     protein_bind    complement(6184..6231)

                     /label=FRT

                     /bound_moiety="FLP recombinase from the Saccharomyces

                     cerevisiae 2u plasmid"

                     /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 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc

     1861 ttcgctatta cgccagctga attggagcga cctcatgcta tacctgagaa agcaacctga

     1921 cctacaggaa agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt

     1981 aaaatttgta tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata

     2041 agaaattcgc ttatttagaa gtgtcaacaa cgtatctacc aacgatttga cccttttcca

     2101 tcttttcgta aatttctggc aaggtagaca agccgacaac cttgattgga gacttgacca

     2161 aacctctggc gaagaattgt taattaagag ctcagatctt atcgtcgtca tccttgtaat

     2221 ccatcgatac tagtgcggcc gccctttagt gagggttgaa ttcgaatttt caaaaattct

     2281 tacttttttt ttggatggac gcaaagaagt ttaataatca tattacatgg cattaccacc

     2341 atatacatat ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa

     2401 gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa

     2461 ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggtgac agccctccga

     2521 aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt

     2581 gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg

     2641 aagaggaaaa attggcagta acctggcccc acaaaccttc aaatgaacga atcaaattaa

     2701 caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc

     2761 gaagcgatga tttttgatct attaacagat atataaatgc aaaaactgca taaccacttt

     2821 aactaatact ttcaacattt tcggtttgta ttacttctta ttcaaatgta ataaaagtat

     2881 caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa accccggatc

     2941 cgtaatacga ctcactatag ggcccgggcg tcgacatgga acagaagttg atttccgaag

     3001 aagacctcga gtaagcttgg taccgcggct agctaagatc cgctctaacc gaaaaggaag

     3061 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa

     3121 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta

     3181 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaagatc cagctgcatt

     3241 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct

     3301 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa

     3361 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa

     3421 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc

     3481 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga

     3541 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc

     3601 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt

     3661 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct

     3721 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg

     3781 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta

     3841 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct

     3901 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa

     3961 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt

     4021 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta

     4081 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat

     4141 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa

     4201 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct

     4261 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta

     4321 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct

     4381 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg

     4441 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa

     4501 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt

     4561 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta

     4621 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca

     4681 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta

     4741 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct

     4801 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg

     4861 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac

     4921 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact

     4981 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa

     5041 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt

     5101 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat

     5161 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg

     5221 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt

     5281 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt

     5341 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa

     5401 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc

     5461 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc

     5521 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg

     5581 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg

     5641 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta

     5701 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat

     5761 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag

     5821 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca

     5881 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa

     5941 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg

     6001 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga

     6061 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc

     6121 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc

     6181 tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt

     6241 ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac

     6301 gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg

     6361 cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct

     6421 agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta

     6481 ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct

     6541 atcatttcct ttgatattgg atcatactaa gaaaccatta ttatcatgac attaacctat

     6601 aaaaataggc gtatcacgag gccctttcgt c

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