| 质粒名称: | SIRT6 Flag |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13817 |
| 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: | 6567 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: | SIRT6 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1067 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: | SIRT6 Flag was a gift from Eric Verdin (爱迪基因 plasmid # 13817) |
| 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 SIRT6 Flag 6486 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression of human SIRT6 with flag tag.
ACCESSION .
VERSION .
KEYWORDS SIRT6 Flag
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6486)
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|SIRT6 Flag|
FEATURES Location/Qualifiers
source 1..6486
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1785..1808
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(1843..1860)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1849..2073
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2119..2547
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2206..2225)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2416..2437
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2556..2576)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2561..2890
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2741..2876
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2803..2822
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2957..3751
/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(3011..3030)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3621..3640
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3925..4046
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3962..3981)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4016..4035
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4095..4111)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4095..4111)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4108..4130)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4119..4135
/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(4143..4173)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4188..4209
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4326..4343)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4497..5085)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4577..4596)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5256..6116)
/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 5879..5898
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6117..6221)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6296..6315)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc gaattccatg tcggtgaatt
721 acgcggcggg gctgtcgccg tacgcggaca agggcaagtg cggcctcccg gagatcttcg
781 accccccgga ggagctggag cggaaggtgt gggaactggc gaggctggtc tggcagtctt
841 ccaatgtggt gttccacacg ggtgccggca tcagcactgc ctctggcatc cccgacttca
901 ggggtcccca cggagtctgg accatggagg agcgaggtct ggcccccaag ttcgacacca
961 cctttgagag cgcgcggccc acgcagaccc acatggcgct ggtgcagctg gagcgcgtgg
1021 gcctcctccg cttcctggtc agccagaacg tggacgggct ccatgtgcgc tcaggcttcc
1081 ccagggacaa actggcagag ctccacggga acatgtttgt ggaagaatgt gccaagtgta
1141 agacgcagta cgtccgagac acagtcgtgg gcaccatggg cctgaaggcc acgggccggc
1201 tctgcaccgt ggctaaggca agggggctgc gagcctgcag gggagagctg agggacacca
1261 tcctagactg ggaggactcc ctgcccgacc gggacctggc actcgccgat gaggccagca
1321 ggaacgccga cctgtccatc acgctgggta catcgctgca gatccggccc agcgggaacc
1381 tgccgctggc taccaagcgc cggggaggcc gcctggtcat cgtcaacctg cagcccacca
1441 agcacgaccg ccatgctgac ctccgcatcc atggctacgt tgacgaggtc atgacccggc
1501 tcatgaagca cctggggctg gagatccccg cctgggacgg cccccgtgtg ctggagaggg
1561 cgctgccacc cctgccccgc ccgcccaccc ccaagctgga gcccaaggag gaatctccca
1621 cccggatcaa cggctctatc cccgccggcc ccaagcagga gccctgcgcc cagcacaacg
1681 gctcagagcc cgccagcccc aaacgggagc ggcccaccag ccctgccccc cacagacccc
1741 ccaaaagggt gaaggccaag gcggtcccca gcgaattcgg aggagactac aaggacgacg
1801 atgacaagta atctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
1861 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
1921 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
1981 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
2041 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
2101 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
2161 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
2221 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
2281 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
2341 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
2401 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
2461 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
2521 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
2581 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
2641 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
2701 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
2761 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
2821 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
2881 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
2941 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
3001 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
3061 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
3121 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
3181 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
3241 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
3301 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
3361 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
3421 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
3481 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
3541 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
3601 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
3661 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
3721 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
3781 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
3841 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
3901 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
3961 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
4021 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
4081 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
4141 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
4201 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
4261 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
4321 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
4381 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
4441 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
4501 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
4561 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
4621 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
4681 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
4741 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
4801 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
4861 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
4921 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
4981 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
5041 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
5101 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
5161 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
5221 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
5281 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
5341 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
5401 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
5461 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
5521 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
5581 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
5641 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
5701 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
5761 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
5821 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
5881 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
5941 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
6001 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
6061 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
6121 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
6181 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
6241 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
6301 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
6361 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
6421 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
6481 cgcgtt
//
