| 质粒名称: | pcDNA3-HA2-Keap1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 21556 |
| 存储实验室: | Yue Xiong |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3-HA2 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 5512 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: | kelch-like ECH-associated protein 1 |
| 物种来源: | -- |
| 插入大小: | 1853 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 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-HA2-Keap1 was a gift from Yue Xiong (Addgene plasmid # 21556 ; http://n2t.net/addgene:21556 ; RRID:Addgene_21556) |
| 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-HA2-Keap1 7314 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3-HA2-Keap1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7314)
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 7314)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7314
/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 2886..2912
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2916..2942
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(4856..4874)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(4857..4874)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(4894..4911)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4900..5124
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 5170..5598
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5257..5276)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5467..5488
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5607..5627)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 5612..5941
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5792..5927
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5854..5873
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 6008..6802
/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(6062..6081)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6672..6691
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6976..7097
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7013..7032)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7067..7086
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(7146..7162)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7146..7162)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7159..7181)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7170..7186
/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(7194..7224)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7239..7260
/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 ctatgtaccc atacgatgtt cctgactatg cgggctatcc ctatgacgtc ccggactatg
2941 caagcttggt accgttaaca tggatcccca tgcagccaga tcccaggcct agcggggctg
3001 gggcctgctg ccgattcctg cccctgcagt cacagtgccc tgagggggca ggggacgcgg
3061 tgatgtacgc ctccactgag tgcaaggcgg aggtgacgcc ctcccagcat ggcaaccgca
3121 ccttcagcta caccctggag gatcatacca agcaggcctt tggcatcatg aacgagctgc
3181 ggctcagcca gcagctgtgt gacgtcacac tgcaggtcaa gtaccaggat gcaccggccg
3241 cccagttcat ggcccacaag gtggtgctgg cctcatccag ccctgtcttc aaggccatgt
3301 tcaccaacgg gctgcgggag cagggcatgg aggtggtgtc cattgagggt atccacccca
3361 aggtcatgga gcgcctcatt gaattcgcct acacggcctc catctccatg ggcgagaagt
3421 gtgtcctcca cgtcatgaac ggtgctgtca tgtaccagat cgacagcgtt gtccgtgcct
3481 gcagtgactt cctggtgcag cagctggacc ccagcaatgc catcggcatc gccaacttcg
3541 ctgagcagat tggctgtgtg gagttgcacc agcgtgcccg ggagtacatc tacatgcatt
3601 ttggggaggt ggccaagcaa gaggagttct tcaacctgtc ccactgccaa ctggtgaccc
3661 tcatcagccg ggacgacctg aacgtgcgct gcgagtccga ggtcttccac gcctgcatca
3721 actgggtcaa gtacgactgc gaacagcgac ggttctacgt ccaggcgctg ctgcgggccg
3781 tgcgctgcca ctcgttgacg ccgaacttcc tgcagatgca gctgcagaag tgcgagatcc
3841 tgcagtccga ctcccgctgc aaggactacc tggtcaagat cttcgaggag ctcaccctgc
3901 acaagcccac gcaggtgatg ccctgccggg cgcccaaggt gggccgcctg atctacaccg
3961 cgggcggcta cttccgacag tcgctcagct acctggaggc ttacaacccc agtgacggca
4021 cctggctccg gttggcggac ctgcaggtgc cgcggagcgg cctggccggc tgcgtggtgg
4081 gcgggctgtt gtacgccgtg ggcggcagga acaactcgcc cgacggcaac accgactcca
4141 gcgccctgga ctgttacaac cccatgacca atcagtggtc gccctgcgcc cccatgagcg
4201 tgccccgtaa ccgcatcggg gtgggggtca tcgatggcca catctatgcc gtcggcggct
4261 cccacggctg catccaccac aacagtgtgg agaggtatga gccagagcgg gatgagtggc
4321 acttggtggc cccaatgctg acacgaagga tcggggtggg cgtggctgtc ctcaatcgtc
4381 tcctttatgc cgtggggggc tttgacggga caaaccgcct taattcagct gagtgttact
4441 acccagagag gaacgagtgg cgaatgatca cagcaatgaa caccatccga agcggggcag
4501 gcgtctgcgt cctgcacaac tgtatctatg ctgctggggg ctatgatggt caggaccagc
4561 tgaacagcgt ggagcgctac gatgtggaaa cagagacgtg gactttcgta gcccccatga
4621 agcaccggcg aagtgccctg gggatcactg tccaccaggg gagaatctac gtccttggag
4681 gctatgatgg tcacacgttc ctggacagtg tggagtgtta cgacccagat acagacacct
4741 ggagcgaggt gacccgaatg acatcgggcc ggagtggggt gggcgtggct gtcaccatgg
4801 agccctgccg gaagcagatt gaccagcaga actgtacctg tctagagggc cctattctat
4861 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
4921 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
4981 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
5041 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
5101 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
5161 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
5221 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
5281 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
5341 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
5401 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
5461 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
5521 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
5581 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
5641 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
5701 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
5761 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
5821 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
5881 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
5941 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
6001 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
6061 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
6121 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
6181 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
6241 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
6301 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
6361 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
6421 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
6481 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
6541 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
6601 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
6661 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
6721 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
6781 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
6841 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
6901 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
6961 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
7021 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
7081 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
7141 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
7201 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
7261 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctg
//
