pUG35

价格:1500元

联系方式:I47-825O-882O

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载体基本信息

出品公司: U.Güldener和J.H.Hegemann
载体名称: pUG35
质粒类型: 酿酒酵母基因敲除载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶
启动子: MET25
载体大小: 6231dp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: C-EGFP
载体抗性: 氨苄
筛选标记: URA3
组成型/诱导型: 诱导型
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息


pUG35载体图谱

载体简介

This vector pUG35 was given by U. GŸldener and J. H. Hegemann from Heinrich-Heine-UniversitŠt in Germany. If a work is published using this vector, proper citation is required! The green fluorescent protein (GFP) of the jellyfish Aequorea victoria has proven to be an extremely useful tool as a marker for gene expression and as a tag to study protein localization. Previously we developed the vectors pGFP-N-FUS and pGFP-C-FUS which allow the N-terminal or C-terminal in-frame fusion of the gfp wildtype gene to the gene of interest and subsequent determination of the subcellular location of the fusion protein. Based on this first set of vectors a new series of GFP vectors equipped with the yEGFP version constructed by Cormack and coworkers . In this yEGFP version all codons of the original gfp gene were exchanged by optimal codons for Candida albicans. In addition 2 amino acid changes were incorporated near the chromophore that had been shown to increase GFP fluorescence . The yEGFP3 not only proved to exhibit stronger fluorescence, but the maturation from the non-fluorescent preform to the mature fluorescence form occurs much faster. The gene encoding the yEGFP3 protein was inserted in the polylinker of the CEN/ARS vectors p413MET25 (HIS3 marker) and p416MET25 (URA3 marker) . Eight (nine in the case of the N-FUS vectors) different restriction sites can be used to insert the gene of interest in frame N-terminal (plasmids pUG23 and pUG35) or C-terminal (plasmids pUG34 and pUG36) to the yEGFP3 encoding open reading frame. The expression level of the fusion protein can be regulated by adding methionine to the media. http://mips.gsf.de/proj/yeast/info/tools/hegemann/gfp.html

载体序列

LOCUS       pUG35                6231 bp    DNA     circular SYN 26-NOV-2000
DEFINITION  C-terminal GFP fusion vector pUG35, complete sequence.
ACCESSION   AF298787
VERSION     AF298787.1
KEYWORDS    .
SOURCE      C-terminal GFP fusion vector pUG35
  ORGANISM  C-terminal GFP fusion vector pUG35
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 6231)
  AUTHORS   Gueldener,U. and Hegemann,J.H.
  TITLE     A second generation of GFP-vectors for subcelluar localization
            studies in budding yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6231)
  AUTHORS   Gueldener,U. and Hegemann,J.H.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2000) Institut fuer Mikrobiologie,
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf
            40225, Germany
FEATURES             Location/Qualifiers
     source          1..6231
                     /organism="C-terminal GFP fusion vector pUG35"
                     /mol_type="other DNA"
                     /db_xref="taxon:142955"
                     /note="derived from Saccharomyces cerevisiae"
     gene            417..1220
                     /gene="URA3"
     CDS             417..1220
                     /gene="URA3"
                     /codon_start=1
                     /transl_table=11
                     /product="orotidine-5'-phosphate decarboxylase"
                     /protein_id="AAG34528.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKEL
                     LELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVK
                     LQYSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLS
                     TGEYTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYR
                     TVDDVVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     regulatory      2004..2262
                     /regulatory_class="terminator"
                     /note="from CYC1"
     gene            complement(2271..2987)
                     /gene="EGFP3"
     CDS             complement(2271..2987)
                     /gene="EGFP3"
                     /codon_start=1
                     /transl_table=11
                     /product="enhanced green fluorescent protein 3"
                     /protein_id="AAG34529.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYK"
     regulatory      3051..3443
                     /regulatory_class="promoter"
                     /note="from MET25"
     rep_origin      3882..3954
     gene            complement(4657..5517)
                     /gene="AmpR"
     CDS             complement(4657..5517)
                     /gene="AmpR"
                     /codon_start=1
                     /transl_table=11
                     /product="ampicillin resistance protein"
                     /protein_id="AAG34530.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     misc_feature    5656..6170
                     /note="CEN6/ARSH4"
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 gccgcaaatt aaagccttcg agcgtcccaa
     2041 aaccttctca agcaaggttt tcagtataat gttacatgcg tacacgcgtc tgtacagaaa
     2101 aaaaagaaaa atttgaaata taaataacgt tcttaatact aacataacta taaaaaaata
     2161 aatagggacc tagacttcag gttgtctaac tccttccttt tcggttagag cggatgtggg
     2221 gggagggcgt gaatgtaagc gtgacataac taattacatg actcgaccag ttatttgtac
     2281 aattcatcca taccatgggt aataccagca gcagtaacaa attctaacaa gaccatgtgg
     2341 tctctctttt cgtttggatc tttggataag gcagattgag tggataagta atggttgtct
     2401 ggtaacaaga ctggaccatc accaattgga gtattttgtt gataatggtc agctaattga
     2461 acagaaccat cttcaatgtt gtgtctaatt ttgaagttaa ctttgatacc attcttttgt
     2521 ttgtcagcca tgatgtaaac attgtgagag ttatagttgt attccaattt gtgacctaaa
     2581 atgttaccat cttctttaaa atcaatacct tttaattcga ttctattaac taaggtatca
     2641 ccttcaaact tgacttcagc tctggtcttg tagttaccgt catctttgaa aaaaatagtt
     2701 ctttcttgaa cataaccttc tggcatggca gacttgaaaa agtcatgttg tttcatatga
     2761 tctgggtatc tagcaaaaca ttgaacacca taaccgaaag tagtgactaa ggttggccat
     2821 ggaactggca atttaccagt agtacaaata aattttaagg tcaatttacc gtaagtagca
     2881 tcaccttcac cttcaccgga gacagaaaat ttgtgaccat taacatcacc atctaattca
     2941 accaaaattg ggacaacacc agtgaataat tcttcacctt tagacatgtc gaggtcgacg
     3001 gtatcgataa gcttgatatc gaattcctgc agcccggggg atccactagt tctagagtat
     3061 ggatgggggt aatagaattg tatctatgta tctgacgacc ctgtattaca cgaaggaaat
     3121 agtagatgaa aagacaagag agcaagaaaa aggaaaaact tcctttctaa cagacgcttt
     3181 acttaacctt atatatatct tatttttttc atcgagcgtg ttcaattgga caaggtgcca
     3241 ccttttcgac acttcggtta ttatgttaca cagttttcat gaggatggcg ccttgactaa
     3301 cttaattagt catttgccaa ccaccacagt tccccaatat cgacagcttc acgtgccatt
     3361 tgccattttg cgatcatgtg acctcaagag aaaaagcaaa aaataatgag agctaaggga
     3421 ttcgaaccct tgcatccgga gctccagctt ttgttccctt tagtgagggt taattgcgcg
     3481 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3541 acacaacata ggagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgaggta
     3601 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3661 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3721 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3781 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3841 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3901 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3961 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4021 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4081 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4141 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4201 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4261 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4321 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4381 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4441 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4501 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4561 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4621 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4681 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4741 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4801 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4861 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4921 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4981 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5041 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5101 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5161 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5221 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5281 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5341 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5401 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5461 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5521 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5581 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5641 gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata
     5701 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     5761 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     5821 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
     5881 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     5941 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     6001 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
     6061 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
     6121 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtcctaa gaaaccatta
     6181 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt c
//					

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