质粒名称: | p112A1NE、p112AINE |
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出品公司: | Addgene |
目录编号: | 85983 |
存储实验室: | Wolf Frommer |
载体骨架基本信息 |
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载体名称: | p112A1NE |
载体类型: | 酵母表达 |
筛选标记: | TRP1 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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启动子: | Adh1 |
克隆方法: | 限制性内切酶 |
文献引用方法 |
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材料和方法部分: | p112A1NE was a gift from Wolf Frommer (Addgene plasmid # 85983 ; http://n2t.net/addgene:85983 ; RRID:Addgene_85983) |
参考文献部分: | Isolation and characterization of a sucrose carrier cDNA from spinach by functional expression in yeast. Riesmeier JW, Willmitzer L, Frommer WB. EMBO J. 1992 Dec;11(13):4705-13. PubMed 1464305 |
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p112AINE载体基本信息
p112AINE载体图谱质粒图谱和多克隆位点信息
p112AINE载体简介
p112AINE载体序列
LOCUS p112A1NE 5725 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For construction of cDNA expression library.. ACCESSION . VERSION . KEYWORDS p112A1NE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5725) AUTHORS Riesmeier JW, Willmitzer L, Frommer WB TITLE Isolation and characterization of a sucrose carrier cDNA from spinach by functional expression in yeast. JOURNAL EMBO J. 1992 Dec;11(13):4705-13. PUBMED 1464305 REFERENCE 2 (bases 1 to 5725) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5725 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 113..132 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 366..383 /label=L4440 /note="L4440 vector, forward primer" protein_bind 500..521 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 536..566 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 574..590 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /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)." primer_bind 579..601 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 598..614 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 598..614 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 638..1034 /gene="S. cerevisiae ADH1" /label=ADH1 promoter /note="promoter for alcohol dehydrogenase 1" terminator 1221..1408 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(1276..1300) /label=pHybLex-R /note="ADH terminator, reverse primer" primer_bind complement(1420..1437) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1420..1436) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1429..1451) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 1873..3219 /label=2u ori /note="yeast 2u plasmid origin of replication" CDS complement(3331..4005) /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /label=TRP1 /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" promoter complement(4006..4107) /gene="S. cerevisiae TRP1" /label=TRP1 promoter primer_bind complement(4127..4145) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4213..4317 /gene="bla" /label=AmpR promoter CDS 4318..5178 /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" primer_bind complement(4536..4555) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5349..212 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 61 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 121 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 181 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt 241 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg 301 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg 361 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc 421 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg 481 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac 541 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag 601 gaaacagcta tgaccatgat tacgccaagc ttgcatgcaa cttcttttct ttttttttct 661 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac 721 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga 781 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc 841 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt 901 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca 961 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa 1021 gcatacaatc aactccaagc ttcagctgct gcagtctaga gaattcccgg gcggccgcgg 1081 atccagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag gttgtcggct 1141 tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg 1201 ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag 1261 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat 1321 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg 1381 tcgctcttat tgaccacacc tctaccggca tgccaattca ctggccgtcg ttttacaacg 1441 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt 1501 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag 1561 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc 1621 acaccgcata tatcggatcg tacttgttac ccatcattga attttgaaca tccgaacctg 1681 ggagttttcc ctgaaacaga tagtatattt gaacctgtat aataatatat agtctagcgc 1741 tttacggaag acaatgtatg tatttcggtt cctggagaaa ctattgcatc tattgcatag 1801 gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt tccatcttgc acttcaatag 1861 catatctttg ttaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga 1921 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg 1981 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg 2041 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca 2101 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg 2161 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg 2221 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga 2281 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt 2341 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata 2401 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc 2461 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag 2521 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt 2581 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat 2641 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat 2701 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag 2761 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt 2821 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcggaatag 2881 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat 2941 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga 3001 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta 3061 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat 3121 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt 3181 ctcatccttc aatgctatca tttcctttga tattggatcg atccgatgat aagctgtcaa 3241 acatgagaat tgatctttta tgcttgcttt tcaaaaggct tgcaggcaag tgcacaaaca 3301 atacttaaat aaatactact cagtaataac ctatttctta gcatttttga cgaaatttgc 3361 tattttgtta gagtctttta caccatttgt ctccacacct ccgcttacat caacaccaat 3421 aacgccattt aatctaagcg catcaccaac attttctggc gtcagtccac cagctaacat 3481 aaaatgtaag ctctcggggc tctcttgcct tccaacccag tcagaaatcg agttccaatc 3541 caaaagttca cctgtcccac ctgcttctga atcaaacaag ggaataaacg aatgaggttt 3601 ctgtgaagct gcactgagta gtatgttgca gtcttttgga aatacgagtc ttttaataac 3661 tggcaaaccg aggaactctt ggtattcttg ccacgactca tctccatgca gttggacgat 3721 atcaatgccg taatcattga ccagagccaa aacatcctcc ttaggttgat tacgaaacac 3781 gccaaccaag tatttcggag tgcctgaact atttttatat gcttttacaa gacttgaaat 3841 tttccttgca ataaccgggt caattgttct ctttctattg ggcacacata taatacccag 3901 caagtcagca tcggaatcta gtgcacattc tgcggcctct gtgctctgca agccgcaaac 3961 tttcaccaat ggaccagaac tacctgtgaa attaataaca gacatactcc aagctgcctt 4021 tgtgtgctta atcacgtata ctcacgtgct caatagtcac caatgccctc cctcttggcc 4081 ctctcctttt cttttttcga ccgaattaat tcttgaagac gaaagggcct cgtgatacgc 4141 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 4201 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 4261 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 4321 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 4381 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 4441 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 4501 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 4561 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 4621 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 4681 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 4741 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 4801 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 4861 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 4921 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 4981 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 5041 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 5101 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 5161 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 5221 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 5281 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 5341 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 5401 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 5461 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 5521 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 5581 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 5641 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 5701 cgaacgacct acaccgaact gagat //
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