质粒名称: | klotho(membrane) |
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出品公司: | 爱迪基因 |
目录编号: | 17712 |
存储实验室: | Hal Dietz |
载体骨架基本信息 |
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载体名称: | pcDNA3.1/V5/His-TOPO |
载体出品公司: | Invitrogen |
载体抗性: | 氨苄霉素 |
空载体大小: | 5500 bp |
载体类型: | 哺乳动物细胞表达 |
筛选标记: | 新霉素(G418) |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Klotho |
物种来源: | 人源 |
GenBank编号 | JN247409 |
启动子 | CMV |
标签/融合蛋白: | V5 (C terminal on backbone) |
克隆方法: | topo 克隆方法 |
5' 测序引物 | CMV-F |
3' 测序引物 | BGH-rev |
文献引用方法 |
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材料和方法部分: | Klotho (membrane) was a gift from Hal Dietz (爱迪基因 plasmid # 17712) |
参考文献部分: |
Association of human aging with a functional variant of klotho. Arking DE, Krebsova A, Macek M, Macek M, Arking A, Mian IS, Fried L, Hamosh A, Dey S, McIntosh I, Dietz HC. Proc Natl Acad Sci U S A. 2002 Jan 22. 99(2):856-61. 10.1073/pnas.022484299 PubMed 11792841 |
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载体基本信息
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载体序列
LOCUS Klotho (membrane) 8575 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Klotho (membrane) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8575) AUTHORS Arking DE, Krebsova A, Macek M, Macek M, Arking A, Mian IS, Fried L, Hamosh A, Dey S, McIntosh I, Dietz HC TITLE Association of human aging with a functional variant of klotho. JOURNAL Proc Natl Acad Sci U S A. 2002 Jan 22. 99(2):856-61. PUBMED 11792841 COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTNAME|klotho(membrane)| FEATURES Location/Qualifiers source 1..8575 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 137..516 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 517..720 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 671..691 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 765..784 /label=T7 /note="T7 promoter, forward primer" promoter 765..783 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 3975..4016 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 4026..4043 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(4066..4083) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4072..4296 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4342..4770 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4429..4448) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4639..4660 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4779..4799) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4784..5113 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4964..5099 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5026..5045 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5180..5974 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5234..5253) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5844..5863 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6150..6271 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6187..6206) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6241..6260 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6320..6336) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6320..6336) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6333..6355) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6344..6360 /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)." promoter complement(6368..6398) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6413..6434 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6551..6568) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6722..7310) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6802..6821) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7481..8341) /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 8104..8123 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8342..8446) /gene="bla" /label=AmpR promoter primer_bind complement(8521..8540) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc 61 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg 121 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg 181 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg 241 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata 301 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc 361 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac 421 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg 481 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc 541 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc 601 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc 661 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct 721 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat 781 agggagaccc aagctggcta gttaagcttg gtaccgagct cggatccact agtccagtgt 841 ggtggaattg cccttcctgg ctcccgcgta gcatgcccgc cagcgccccg ccgcgccgcc 901 cgcggccgcc gccgccgtcg ctgtcgctgc tgctggtgct gctgggcctg ggcggccgcc 961 gcctgcgtgc ggagccgggc gacggcgcgc agacctgggc ccgtttctcg cggcctcctg 1021 cccccgaggc cgcgggcctc ttccagggca ccttccccga cggcttcctc tgggccgtgg 1081 gcagcgccgc ctaccagacc gagggcggct ggcagcagca cggcaagggt gcgtccatct 1141 gggacacgtt cacccaccac cccctggcac cctcgggaga ctcccggaac gccagtctgc 1201 cgttgggcgc cccgtcgccg ctgcagcccg ccaccgggga cgtagccagc gacagctaca 1261 acaacgtctt ccgcgacacg gaggcgctgc gcgagctcgg ggtcactcac taccgcttct 1321 ccatctcgtg ggcgcgagtg ctccccaatg gcagcgcggg cgtccccaac cgcgaggggc 1381 tgcgctacta ccggcgcctg ctggagcggc tgcgggagct gggcgtgcag cccgtggtca 1441 ccctgtacca ctgggacctg ccccagcgcc tgcaggacgc ctatggcggc tgggccaacc 1501 gcgccctggc cgaccacttc agggattacg cggagctctg cttccgccac ttcggcggtc 1561 aggtcaagta ctggatcacc atcgacaacc cctacgtggt ggcctggcac ggctacgcca 1621 ccgggcgcct ggcccccggc atccggggca gcccgcggct cgggtacctg gtggcgcaca 1681 acctcctcct ggctcatgcc aaagtctggc atctctacaa tacttctttc cgtcccactc 1741 agggaggtca ggtgtccatt gccctaagct ctcactggat caatcctcga agaatgaccg 1801 accacagcat caaagaatgt caaaaatctc tggactttgt actaggttgg tttgccaaac 1861 ccgtatttat tgatggtgac tatcccgaga gcatgaagaa taacctttca tctattctgc 1921 ctgattttac tgaatctgag aaaaagttca tcaaaggaac tgctgacttt tttgctcttt 1981 gctttggacc caccttgagt tttcaacttt tggaccctca catgaagttc cgccaattgg 2041 aatctcccaa cctgaggcaa ctgctttcct ggattgacct tgaatttaac catcctcaaa 2101 tatttattgt ggaaaatggc tggtttgtct cagggaccac caagagagat gatgccaaat 2161 atatgtatta cctcaaaaag ttcatcatgg aaaccttaaa agccatcaag ctggatgggg 2221 tggatgtcat cgggtatacc gcatggtccc tcatggatgg tttcgagtgg cacagaggtt 2281 acagcatcag gcgtggactc ttctatgttg actttctaag ccaggacaag atgttgttgc 2341 caaagtcttc agccttgttc taccaaaagc tgatagagaa aaatggcttc cctcctttac 2401 ctgaaaatca gcccctagaa gggacatttc cctgtgactt tgcttgggga gttgttgaca 2461 actacattca agtagatacc actctgtctc agtttaccga cctgaatgtt tacctgtggg 2521 atgtccacca cagtaaaagg cttattaaag tggatggggt tgtgaccaag aagaggaaat 2581 cctactgtgt tgactttgct gccatccagc cccagatcgc tttactccag gaaatgcacg 2641 ttacacattt tcgcttctcc ctggactggg ccctgattct ccctctgggt aaccagtccc 2701 aggtgaacca caccatcctg cagtactatc gctgcatggc cagcgagctt gtccgtgtca 2761 acatcacccc agtggtggcc ctgtggcagc ctatggcccc gaaccaagga ctgccgcgcc 2821 tcctggccag gcagggcgcc tgggagaacc cctacactgc cctggccttt gcagagtatg 2881 cccgactgtg ctttcaagag ctcggccatc acgtcaagct ttggataacg atgaatgagc 2941 cgtatacaag gaatatgaca tacagtgctg gccacaacct tctgaaggcc catgccctgg 3001 cttggcatgt gtacaatgaa aagtttaggc atgctcagaa tgggaaaata tccatagcct 3061 tgcaggctga ttggatagaa cctgcctgcc ctttctccca aaaggacaaa gaggtggctg 3121 agagagtttt ggaatttgac attggctggc tggctgagcc cattttcggc tctggagatt 3181 atccatgggt gatgagggac tggctgaacc aaagaaacaa ttttcttctt ccttatttca 3241 ctgaagatga aaaaaagcta atccagggta cctttgactt tttggcttta agccattata 3301 ccaccatcct tgtagactca gaaaaagaag atccaataaa atacaatgat tacctagaag 3361 tgcaagaaat gaccgacatc acgtggctca actcccccag tcaggtggcg gtagtgccct 3421 gggggttgcg caaagtgctg aactggctga agttcaagta cggagacctc cccatgtaca 3481 taatatccaa cggaatcgat gacgggctgc atgctgagga cgaccagctg agggtgtatt 3541 atatgcagaa ttacataaac gaagctctca aagcccacat actggatggt atcaatcttt 3601 gcggatactt tgcttattcg tttaacgacc gcacagctcc gaggtttggc ctctatcgtt 3661 atgctgcaga tcagtttgag cccaaggcat ccatgaaaca ttacaggaaa attattgaca 3721 gcaatggttt cccgggccca gaaactctgg aaagattttg tccagaagaa ttcaccgtgt 3781 gtactgagtg cagttttttt cacacccgaa agtctttact ggctttcata gcttttctat 3841 tttttgcttc tattatttct ctctccctta tattttacta ctcgaagaaa ggcagaagaa 3901 gttacaaaaa gggcaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg 3961 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta 4021 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt 4081 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg 4141 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt 4201 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca 4261 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct 4321 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg 4381 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt 4441 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc 4501 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg 4561 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg 4621 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct 4681 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg 4741 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg 4801 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 4861 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 4921 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc 4981 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 5041 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 5101 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac 5161 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 5221 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 5281 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 5341 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 5401 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 5461 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 5521 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 5581 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 5641 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 5701 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 5761 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 5821 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 5881 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 5941 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga 6001 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 6061 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 6121 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 6181 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 6241 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 6301 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6361 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6421 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 6481 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 6541 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 6601 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 6661 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 6721 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 6781 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 6841 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 6901 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 6961 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7021 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7081 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7141 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 7201 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 7501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 7561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 7621 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 7681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 7741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 7801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 7861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 7921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 7981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 8461 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag 8521 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtg //
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