| 质粒名称: | pcDNA3-EGFP-C4-Nrf2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 21549 |
| 存储实验室: | Yue Xiong |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 6112 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 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: | nuclear factor (erythroid-derived 2)-like 2 |
| 物种来源: | -- |
| 插入大小: | 1825bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3-EGFP-C4-Nrf2 was a gift from Yue Xiong (Addgene plasmid # 21549 ; http://n2t.net/addgene:21549 ; RRID:Addgene_21549) |
| 参考文献部分: | BTB protein Keap1 targets antioxidant transcription factor Nrf2 for ubiquitination by the Cullin 3-Roc1 ligase. Furukawa M, Xiong Y. Mol Cell Biol. 2005 Jan . 25(1):162-71. 10.1128/MCB.25.1.162-171.2005 PubMed 15601839 |
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 pcDNA3-EGFP-C4-Nrf2 7972 bp ds-DNA circular SYN 22-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3-EGFP-C4-Nrf2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7972)
AUTHORS Furukawa M, Xiong Y
TITLE BTB protein Keap1 targets antioxidant transcription factor Nrf2 for
ubiquitination by the Cullin 3-Roc1 ligase.
JOURNAL Mol Cell Biol. 2005 Jan . 25(1):162-71.
PUBMED 15601839
REFERENCE 2 (bases 1 to 7972)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 22, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7972
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 51..430
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 431..634
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 585..605
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 679..698
/label=T7
/note="T7 promoter, forward primer"
promoter 679..697
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 723..732
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 729..1445
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(774..795)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1035..1054)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1382..1403
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1464..1493
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
promoter complement(3341..3359)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3342..3359)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(3379..3396)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3385..3609
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3655..4083
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3742..3761)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3952..3973
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4092..4112)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4097..4426
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4277..4412
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4339..4358
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4493..5287
/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(4547..4566)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5157..5176
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5461..5582
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5498..5517)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5552..5571
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5631..5647)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5631..5647)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5644..5666)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5655..5671
/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(5679..5709)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5724..5745
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5862..5879)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6033..6621)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6113..6132)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6792..7652)
/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 7415..7434
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7653..7757)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7832..7851)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
61 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
121 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
181 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
361 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
421 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
481 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
541 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
601 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
661 ttactggctt atcgaaatta atacgactca ctatagggag acccaagcta gcgctaccgg
721 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
781 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
841 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
901 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
961 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
1021 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
1081 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
1141 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
1201 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
1261 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
1321 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
1381 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
1441 acaagtactc agatctggaa ttagagcaaa agctcatctc agaggaagat ctggaattca
1501 ggatcctcat gatggacttg gagctgccgc cgccgggact cccgtcccag caggacatgg
1561 atttgattga catactttgg aggcaagata tagatcttgg agtaagtcga gaagtatttg
1621 acttcagtca gcgacggaaa gagtatgagc tggaaaaaca gaaaaaactt gaaaaggaaa
1681 gacaagaaca actccaaaag gagcaagaga aagccttttt cgctcagtta caactagatg
1741 aagagacagg tgaatttctc ccaattcagc cagcccagca catccagtca gaaaccagtg
1801 gatctgccaa ctactcccag gttgcccaca ttcccaaatc agatgctttg tactttgatg
1861 actgcatgca gcttttggcg cagacattcc cgtttgtaga tgacaatgag gtttcttcgg
1921 ctacgtttca gtcacttgtt cctgatattc ccggtcacat cgagagccca gtcttcattg
1981 ctactaatca ggctcagtca cctgaaactt ctgttgctca ggtagcccct gttgatttag
2041 acggtatgca acaggacatt gagcaagttt gggaggagct attatccatt cctgagttac
2101 agtgtcttaa tattgaaaat gacaagctgg ttgagactac catggttcca agtccagaag
2161 ccaaactgac agaagttgac aattatcatt tttactcatc tataccctca atggaaaaag
2221 aagtaggtaa ctgtagtcca cattttctta atgcttttga ggattccttc agcagcatcc
2281 tctccacaga agaccccaac cagttgacag tgaactcatt aaattcagat gccacagtca
2341 acacagattt tggtgatgaa ttttattctg ctttcatagc tgagcccagt atcagcaaca
2401 gcatgccctc acctgctact ttaagccatt cactctctga acttctaaat gggcccattg
2461 atgtttctga tctatcactt tgcaaagctt tcaaccaaaa ccaccctgaa agcacagcag
2521 aattcaatga ttctgactcc ggcatttcac taaacacaag tcccagtgtg gcatcaccag
2581 aacactcagt ggaatcttcc agctatggag acacactact tggcctcagt gattctgaag
2641 tggaagagct agatagtgcc cctggaagtg tcaaacagaa tggtcctaaa acaccagtac
2701 attcttctgg ggatatggta caacccttgt caccatctca ggggcagagc actcacgtgc
2761 atgatgccca atgtgagaac acaccagaga aagaattgcc tgtaagtcct ggtcatcgga
2821 aaaccccatt cacaaaagac aaacattcaa gccgcttgga ggctcatctc acaagagatg
2881 aacttagggc aaaagctctc catatcccat tccctgtaga aaaaatcatt aacctccctg
2941 ttgttgactt caacgaaatg atgtccaaag agcagttcaa tgaagctcaa cttgcattaa
3001 ttcgggatat acgtaggagg ggtaagaata aagtggctgc tcagaattgc agaaaaagaa
3061 aactggaaaa tatagtagaa ctagagcaag atttagatca tttgaaagat gaaaaagaaa
3121 aattgctcaa agaaaaagga gaaaatgaca aaagccttca cctactgaaa aaacaactca
3181 gcaccttata tctcgaagtt ttcagcatgc tacgtgatga agatggaaaa ccttattctc
3241 ctagtgaata ctccctgcag caaacaagag atggcaatgt tttccttgtt cccaaaagta
3301 agaagccaga tgttaagaaa aactatctag agggccctat tctatagtgt cacctaaatg
3361 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
3421 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
3481 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
3541 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
3601 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
3661 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
3721 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
3781 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
3841 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
3901 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
3961 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
4021 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
4081 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
4141 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
4201 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
4261 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
4321 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
4381 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
4441 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
4501 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
4561 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
4621 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
4681 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
4741 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
4801 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
4861 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
4921 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
4981 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
5041 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
5101 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
5161 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
5221 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
5281 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
5341 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
5401 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
5461 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
5521 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
5581 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
5641 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
5701 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
5761 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
5821 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
5881 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
5941 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
6001 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
6061 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
6121 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
6181 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
6241 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
6301 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
6361 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
6421 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
6481 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
6541 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
6601 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
6661 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
6721 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
6781 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
6841 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
6901 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
6961 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
7021 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
7081 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
7141 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
7201 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
7261 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
7321 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
7381 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
7441 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
7501 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
7561 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
7621 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
7681 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
7741 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
7801 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
7861 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
7921 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt ag
//
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