| 质粒名称: | SIRT5 Flag |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13816 |
| 存储实验室: | Eric Verdin |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 + |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5500 bp |
| 完整质粒大小: | 6432 bp |
| 载体类型: | 哺乳细胞表达 |
| 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 |
|
| 插入基因名称: | SIRT5 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 932 bp |
| 标签/融合蛋白: | flag (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | EcoRI(未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | BGH Reverse |
|
文献引用方法 |
|
| 材料和方法部分: | SIRT5 Flag was a gift from Eric Verdin (爱迪基因 plasmid # 13816) |
| 参考文献部分: | The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin deacetylase. North BJ, Marshall BL, Borra MT, Denu JM, Verdin E. Mol Cell. 2003 Feb . 11(2):437-44. 10.1016/S1097-2765(03)00038-8 PubMed 12620231 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS SIRT5 Flag 6351 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression of human SIRT5 with flag tag.
ACCESSION .
VERSION .
KEYWORDS SIRT5 Flag
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6351)
AUTHORS North BJ, Marshall BL, Borra MT, Denu JM, Verdin E
TITLE The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin
deacetylase.
JOURNAL Mol Cell. 2003 Feb . 11(2):437-44.
PUBMED 12620231
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|SIRT5 Flag|
FEATURES Location/Qualifiers
source 1..6351
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(63..80)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(234..822)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(314..333)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(993..1853)
/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 1616..1635
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1854..1958)
/gene="bla"
/label=AmpR promoter
primer_bind complement(2033..2052)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 2224..2603
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2604..2807
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2758..2778
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2852..2871
/label=T7
/note="T7 promoter, forward primer"
promoter 2852..2870
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 3873..3896
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(3931..3948)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3937..4161
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4207..4635
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4294..4313)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4504..4525
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4644..4664)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4649..4978
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4829..4964
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4891..4910
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5045..5839
/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(5099..5118)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5709..5728
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6013..6134
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6050..6069)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6104..6123
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6183..6199)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6183..6199)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6196..6218)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6207..6223
/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(6231..6261)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6276..6297
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
2881 ctggctagcg tttaaactta agcttggtac cgagctcgga tccgaattcc atgcgacctc
2941 tccagattgt cccaagtcga ttgatttccc agctatattg tggcctgaag cctccagcgt
3001 ccacacgaaa ccagatttgc ctgaaaatgg ctcggccaag ttcaagtatg gcagattttc
3061 gaaagttttt tgcaaaagca aagcacatag tcatcatctc aggagctggt gttagtgcag
3121 aaagtggtgt tccgaccttc agaggagctg gaggttattg gagaaaatgg caagcccagg
3181 acctggcgac tcccctggcc tttgcccaca acccgtcccg ggtgtgggag ttctaccact
3241 accggcggga ggtcatgggg agcaaggagc ccaacgccgg gcaccgcgcc atagccgagt
3301 gtgagacccg gctgggcaag cagggccggc gagtcgtggt catcacccag aacatcgatg
3361 agctgcaccg caaggctggc accaagaacc ttctggagat ccatggtagc ttatttaaaa
3421 ctcgatgtac ctcttgtgga gttgtggctg agaattacaa gagtccaatt tgtccagctt
3481 tatcaggaaa aggtgctcca gaacctggaa ctcaagatgc cagcatccca gttgagaaac
3541 ttccccggtg tgaagaggca ggctgcgggg gcttgctgcg acctcacgtc gtgtggtttg
3601 gagaaaacct ggatcctgcc attctggagg aggttgacag agagctcgcc cactgtgatt
3661 tatgtctagt ggtgggcact tcctctgtgg tgtacccagc agccatgttt gccccccagg
3721 tggctgccag gggcgtgcca gtggctgaat ttaacacgga gaccacccca gctacgaaca
3781 gattcaggtt tcatttccag ggaccctgtg gaacgactct tcctgaagcc cttgcctgtc
3841 atgaaaatga aactgtttct gaattcggag gagactacaa ggacgacgat gacaagtaat
3901 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
3961 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
4021 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
4081 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
4141 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
4201 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
4261 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
4321 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
4381 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
4441 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
4501 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
4561 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
4621 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
4681 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
4741 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
4801 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
4861 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
4921 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
4981 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
5041 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
5101 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
5161 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
5221 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
5281 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
5341 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
5401 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
5461 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
5521 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
5581 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
5641 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
5701 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
5761 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
5821 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
5881 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
5941 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
6001 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
6061 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
6121 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
6181 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
6241 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
6301 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct g
//
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