| 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 |
买家导航
pUG35载体质粒基本信息
pUG35质粒图谱载体图谱和pUG35载体序列质粒序列多克隆位点信息
pUG35质粒载体简介
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
pUG35质粒序列载体序列
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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pUG35质粒载体应用举例
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