| 質粒名稱: | Klotho(secreted) |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 17713 |
| 存儲實驗室: | Hal Dietz |
|
載體骨架基本信息 |
|
| 載體名稱: | pcDNA3.1/V5/His-TOPO |
| 載體出品公司: | Invitrogen |
| 載體抗性: | 氨芐霉素 |
| 空載體大小: | 5500 bp |
| 載體類型: | 哺乳細胞表達 |
| 篩選標記: | 新霉素（G418） |
| 高拷貝/低拷貝: | 高拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | Klotho |
| 物種來源: | 人源 |
| GenBank編號 | JN247409 |
| 克隆方法: | TOPO Cloning |
| 5' 測序引物 | CMV-F |
| 3' 測序引物 | BGH-rev |
|
文獻引用方法 |
|
| 材料和方法部分: | Klotho (secreted) was a gift from Hal Dietz (Addgene plasmid # 17713) |
| 參考文獻部分: |
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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Klotho(secreted)載體質粒基本信息
Klotho(secreted)質粒圖譜載體圖譜和Klotho(secreted)載體序列質粒序列多克隆位點信息
載體圖譜.png)
Klotho(secreted)質粒載體簡介
Klotho(secreted)質粒序列載體序列
LOCUS Klotho(secreted) 7186 bp DNA circular SYN 14-APR-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7186)
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
REFERENCE 2 (bases 1 to 7186)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A. 2002 Jan 22. 99(2):856-61."
COMMENT Sequence Label: Klotho (secreted)
FEATURES Location/Qualifiers
source 1..7186
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2684..2725
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 2735..2752
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2775..2792)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2781..3005
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3051..3479
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3138..3157)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3348..3369
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3488..3508)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3493..3822
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3673..3808
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3735..3754
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3889..4683
/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(3943..3962)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4553..4572
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4859..4980
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4896..4915)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4950..4969
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5029..5045)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5029..5045)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5042..5064)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5053..5069
/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(join(5077..5083,5084..5101,5102..5107))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5122..5143
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5260..5277)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5431..6019)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5511..5530)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(6190..6981,6982..7050))
/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 6813..6832
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7051..7155)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattgcc cttcctggct
961 cccgcgcagc atgcccgcca gcgccccgcc gcgccgcccg cggccgccgc cgccgtcgct
1021 gtcgctgctg ctggtgctgc tgggcctggg cggccgccgc ctgcgtgcgg agccgggcga
1081 cggcgcgcag acctgggccc gtttctcgcg gcctcctgcc cccgaggccg cgggcctctt
1141 ccagggcacc ttccccgacg gcttcctctg ggccgtgggc agcgccgcct accagaccga
1201 gggcggctgg cagcagcacg gcaagggtgc gtccatctgg gacacgttca cccaccaccc
1261 cctggcaccc ccgggagact cccggaacgc cagtctgccg ttgggcgccc cgtcgccgct
1321 gcagcccgcc accggggacg tagccagcga cagctacaac aacgtcttcc gcgacacgga
1381 ggcgctgcgc gagctcgggg tcactcacta ccgcttctcc atctcgtggg cgcgagtgct
1441 ccccaatggc agcgcgggcg tccccaaccg cgaggggctg cgctactacc ggcgcctgct
1501 ggagcggctg cgggagctgg gcgtgcagcc cgtggtcacc ctgtaccact gggacctgcc
1561 ccagcgcctg caggacgcct acggcggctg ggccaaccgc gccctggccg accacttcag
1621 ggattacgcg gagctctgct tccgccactt cggcggtcag gtcaagtact ggatcaccat
1681 cgacaacccc tacgtggtgg cctggcacgg ctacgccacc gggcgcctgg cccccggcat
1741 ccggggcagc ccgcggctcg ggtacctggt ggcgcacaac ctcctcctgg ctcatgccaa
1801 agtctggcat ctctacaata cttctttccg tcccactcag ggaggtcagg tgtccattgc
1861 cctaagctct cactggatca atcctcgaag aatgaccgac cacagcatca aagaatgtca
1921 aaaatctctg gactttgtac taggttggtt tgccaaaccc gtatttattg atggtgacta
1981 tcccgagagc atgaagaata acctttcatc tattctgcct gattttactg aatctgagaa
2041 aaagttcatc aaaggaactg ctgacttttt tgctctttgc tttggaccca ccttgagttt
2101 tcaacttttg gaccctcaca tgaagttccg ccaattggaa tctcccaacc tgaggcaact
2161 gctttcctgg attgaccttg aatttaacca tcctcaaata tttattgtgg aaaatggctg
2221 gtttgtctca gggaccacca agagagatga tgccaaatat atgtattacc tcaaaaagtt
2281 catcatggaa accttaaaag ccatcaagct ggatggggtg gatgtcatcg ggtataccgc
2341 atggtccctc atggatggtt tcgagtggca cagaggttac agcatcaggc gtggactctt
2401 ctatgttgac tttctaagcc aggacaagat gttgttgcca aagtcttcag ccttgttcta
2461 ccaaaagctg atagagaaaa atggcttccc tcccttacct gaaaatcagc ccctagaagg
2521 gacatttccc tgtgactttg cttggggagt tgttgacaac tacattcaag taagtcagct
2581 gacaaaacca atcagcagtc tcaccaagcc ctatcacaag ggcaattctg cagatatcca
2641 gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc ctatccctaa
2701 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta
2761 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
2821 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
2881 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
2941 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
3001 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
3061 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
3121 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
3181 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
3241 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
3301 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
3361 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
3421 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
3481 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
3541 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
3661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
3721 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
3781 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
3841 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
3901 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
3961 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
4021 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
4081 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
4141 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
4201 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
4261 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
4321 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
4381 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
4441 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
4501 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
4561 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
4621 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
4681 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
4741 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
4801 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
4861 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
4921 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
4981 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
5041 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
5101 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
5161 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
5221 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
5281 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
5341 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
5401 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
5461 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
5521 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
5581 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
5641 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
5701 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
5761 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
5821 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
5881 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
5941 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
6001 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
6061 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
6121 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
6181 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
6241 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
6301 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
6361 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
6421 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
6481 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
6541 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
6601 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
6661 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
6721 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
6781 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
6841 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
6901 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
6961 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
7021 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
7081 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
7141 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
Klotho(secreted)載體圖譜質粒圖譜pdf版和Klotho(secreted)載體序列質粒序列等相關資料下載
Klotho(secreted)質粒載體應用舉例
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