| 质粒名称: | SIRT3 Flag |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13814 |
| 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: | 6699 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: | SIRT3 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1199 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: | SIRT3 Flag was a gift from Eric Verdin (爱迪基因 plasmid # 13814) |
| 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 |
SIRT3 Flag
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LOCUS SIRT3 Flag 6618 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression of human SIRT3 with flag tag.
ACCESSION .
VERSION .
KEYWORDS SIRT3 Flag
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6618)
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|SIRT3 Flag|
FEATURES Location/Qualifiers
source 1..6618
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(11..28)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(182..770)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(262..281)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(941..1801)
/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 1564..1583
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1802..1906)
/gene="bla"
/label=AmpR promoter
primer_bind complement(1981..2000)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 2172..2551
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2552..2755
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2706..2726
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2800..2819
/label=T7
/note="T7 promoter, forward primer"
promoter 2800..2818
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 4088..4111
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(4146..4163)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4152..4376
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4422..4850
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4509..4528)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4719..4740
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4859..4879)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4864..5193
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5044..5179
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5106..5125
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5260..6054
/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(5314..5333)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5924..5943
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6228..6349
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6265..6284)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6319..6338
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6398..6414)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6398..6414)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6411..6433)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6422..6438
/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(6446..6476)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6491..6512
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
2821 gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc cgaattccat
2881 ggcgttctgg ggttggcgcg ccgcggcagc cctccggctg tggggccggg tagttgaacg
2941 ggtcgaggcc gggggaggcg tggggccgtt tcaggcctgc ggctgtcggc tggtgcttgg
3001 cggcagggac gatgtgagtg cggggctgag aggcagccat ggggcccgcg gtgagccctt
3061 ggacccggcg cgccccttgc agaggcctcc cagacccgag gtgcccaggg cattccggag
3121 gcagccgagg gcagcagctc ccagtttctt cttttcgagt attaaaggtg gaagaaggtc
3181 catatctttt tctgtgggtg cttcaagtgt tgttggaagt ggaggcagca gtgacaaggg
3241 gaagctttcc ctgcaggatg tagctgagct gattcgggcc agagcctgcc agagggtggt
3301 ggtcatggtg ggggccggca tcagcacacc cagtggcatt ccagacttca gatcgccggg
3361 gagtggcctg tacagcaacc tccagcagta cgatctcccg taccccgagg ccatttttga
3421 actcccattc ttctttcaca accccaagcc ctttttcact ttggccaagg agctgtaccc
3481 tggaaactac aagcccaacg tcactcacta ctttctccgg ctgcttcatg acaaggggct
3541 gcttctgcgg ctctacacgc agaacatcga tgggcttgag agagtgtcgg gcatccctgc
3601 ctcaaagctg gttgaagctc atggaacctt tgcctctgcc acctgcacag tctgccaaag
3661 acccttccca ggggaggaca ttcgggctga cgtgatggca gacagggttc cccgctgccc
3721 ggtctgcacc ggcgttgtga agcccgacat tgtgttcttt ggggagccgc tgccccagag
3781 gttcttgctg catgtggttg atttccccat ggcagatctg ctgctcatcc ttgggacctc
3841 cctggaggtg gagccttttg ccagcttgac cgaggccgtg cggagctcag ttccccgact
3901 gctcatcaac cgggacttgg tggggccctt ggcttggcat cctcgcagca gggacgtggc
3961 ccagctgggg gacgtggttc acggcgtgga aagcctagtg gagcttctgg gctggacaga
4021 agagatgcgg gaccttgtgc agcgggaaac tgggaagctt gatggaccag acaaagaatt
4081 cggaggagac tacaaggacg acgatgacaa gtaatctaga gggcccgttt aaacccgctg
4141 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
4201 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
4261 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
4321 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
4381 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
4441 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
4501 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
4561 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
4621 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
4681 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
4741 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
4801 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
4861 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
4921 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
4981 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
5041 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
5101 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
5161 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
5221 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
5281 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
5341 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
5401 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
5461 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
5521 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
5581 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
5641 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
5701 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
5761 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
5821 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
5881 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
5941 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
6001 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
6061 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
6121 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
6181 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
6241 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
6301 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
6361 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
6421 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
6481 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
6541 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
6601 gtttgcgtat tgggcgct
//
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