| 质粒名称: | pGL4.27_ARE NRF2-SPE |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 177775 |
| 存储实验室: | Michael Ristow |
|
载体骨架基本信息 |
|
| 载体名称: | pGL4.27 |
| 完整质粒大小: | 6072 bp |
| 载体类型: | 荧光素酶 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | NRF2 |
| 物种来源: | H. sapiens (human) |
| 基因突变: | NFE2L2 (a.k.a. HEBP1, IMDDHH, NRF2, Nrf-2) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pGL4.27_ARE/NRF2-SPE was a gift from Michael Ristow (Addgene plasmid # 177775 ; http://n2t.net/addgene:177775 ; RRID:Addgene_177775) |
| 参考文献部分: | Grainyhead 1 acts as a drug-inducible conserved transcriptional regulator linked to insulin signaling and lifespan. Grigolon G, Araldi E, Erni R, Wu JY, Thomas C, La Fortezza M, Laube B, Pohlmann D, Stoffel M, Zarse K, Carreira EM, Ristow M, Fischer F. Nat Commun. 2022 Jan 10;13(1):107. doi: 10.1038/s41467-021-27732-4. 10.1038/s41467- |
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 pGL4.27_ARE NRF2-SPE 6072 bp ds-DNA circular SYN 15-FEB-2022
DEFINITION Luciferase reporter for ARE/NRF2.
ACCESSION .
VERSION .
KEYWORDS pGL4.27_ARE/NRF2-SPE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6072)
TITLE Ristow lab plasmids
REFERENCE 2 (bases 1 to 6072)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 15, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6072
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(1..861)
/codon_start=1
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
polyA_signal 966..1014
/note="synthetic polyadenylation signal"
misc_feature 1028..1119
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 1068..1087
/label=RVprimer3
/note="pGL3 vector, forward primer"
promoter 1258..1289
/label=minP
/note="minimal TATA-box promoter with low basal activity"
regulatory 1316..1325
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1322..2971
/codon_start=1
/product="firefly luciferase"
/label=luciferase
/note="synthetic luc2 version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
CDS 2975..3094
/codon_start=1
/product="PEST degradation sequence from mouse ornithine
decarboxylase"
/label=hPEST
/note="human codon-optimized"
/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
polyA_signal 3146..3267
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3157..3176)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3211..3230
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 3461..3818
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3484..3504)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3669..3804
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3731..3750
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3849..4886
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
EVGRV"
RBS 4865..4873
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
polyA_signal 4910..4958
/note="synthetic polyadenylation signal"
primer_bind complement(5114..5131)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5285..5873)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5365..5384)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat
61 agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc
121 cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa
181 ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca
241 gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag
301 agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt
361 caactctggt tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc
421 agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct
481 catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc
541 cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg
601 ctcttgcccg gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct
661 catcatcggg aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc
721 cagttcgata tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag
781 cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac
841 acgaaaatgt tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg
901 ttactagtac gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag
961 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
1021 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
1081 ctgtccccag tgcaagtgca ggtgccagaa catttctctg gcctaactgg ccggtacctc
1141 acagtgactc agcatttaaa cacagtgact cagcatttaa acacagtgac tcagcattaa
1201 agagctcgct agcctcgagg atatcaagat ctggcctcgg cggccaagct tagacactag
1261 agggtatata atggaagctc gacttccagc ttggcaatcc ggtactgttg gtaaagccac
1321 catggaagat gccaaaaaca ttaagaaggg cccagcgcca ttctacccac tcgaagacgg
1381 gaccgccggc gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcaccat
1441 cgcctttacc gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag
1501 cgttcggctg gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt
1561 gtgcagcgag aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt
1621 ggctgtggcc ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat
1681 cagccagccc accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca
1741 aaagaagcta ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg
1801 cttccaaagc atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga
1861 cttcgtgccc gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg
1921 cagtaccgga ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag
1981 tcatgcccgc gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt
2041 ggtgccattt caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt
2101 tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta
2161 taagattcaa tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct
2221 catcgacaag tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag
2281 caaggaggta ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta
2341 cggcctgaca gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg
2401 cgcagtaggc aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa
2461 gacactgggt gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg
2521 ctacgttaac aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag
2581 cggcgacatc gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag
2641 cctgatcaaa tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca
2701 acaccccaac atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct
2761 gcccgccgca gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga
2821 ctatgtggcc agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga
2881 cgaggtgcct aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat
2941 taaggccaag aagggcggca agatcgccgt gaattctcac ggcttccctc ccgaggtgga
3001 ggagcaggcc gccggcaccc tgcccatgag ctgcgcccag gagagcggca tggatagaca
3061 ccctgctgct tgcgccagcg ccaggatcaa cgtctaaggc cgcgactcta gagtcggggc
3121 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
3181 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
3241 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
3301 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
3361 atcgataagg atccgtttgc gtattgggcg ctcttccgct gatctgcgca gcaccatggc
3421 ctgaaataac ctctgaaaga ggaacttggt tagctacctt ctgaggcgga aagaaccagc
3481 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
3541 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
3601 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
3661 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
3721 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
3781 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgattct tctgacacta
3841 gcgccaccat gaagaagccc gaactcaccg ctaccagcgt tgaaaaattt ctcatcgaga
3901 agttcgacag tgtgagcgac ctgatgcagt tgtcggaggg cgaagagagc cgagccttca
3961 gcttcgatgt cggcggacgc ggctatgtac tgcgggtgaa tagctgcgct gatggcttct
4021 acaaagaccg ctacgtgtac cgccacttcg ccagcgctgc actacccatc cccgaagtgt
4081 tggacatcgg cgagttcagc gagagcctga catactgcat cagtagacgc gcccaaggcg
4141 ttactctcca agacctcccc gaaacagagc tgcctgctgt gttacagcct gtcgccgaag
4201 ctatggatgc tattgccgcc gccgacctca gtcaaaccag cggcttcggc ccattcgggc
4261 cccaaggcat cggccagtac acaacctggc gggatttcat ttgcgccatt gctgatcccc
4321 atgtctacca ctggcagacc gtgatggacg acaccgtgtc cgccagcgta gctcaagccc
4381 tggacgaact gatgctgtgg gccgaagact gtcccgaggt gcgccacctc gtccatgccg
4441 acttcggcag caacaacgtc ctgaccgaca acggccgcat caccgccgta atcgactggt
4501 ccgaagctat gttcggggac agtcagtacg aggtggccaa catcttcttc tggcggccct
4561 ggctggcttg catggagcag cagactcgct acttcgagcg ccggcatccc gagctggccg
4621 gcagccctcg tctgcgagcc tacatgctgc gcatcggcct ggatcagctc taccagagcc
4681 tcgtggacgg caacttcgac gatgctgcct gggctcaagg ccgctgcgat gccatcgtcc
4741 gcagcggggc cggcaccgtc ggtcgcacac aaatcgctcg ccggagcgca gccgtatgga
4801 ccgacggctg cgtcgaggtg ctggccgaca gcggcaaccg ccggcccagt acacgaccgc
4861 gcgctaagga ggtaggtcga gtttaaactc tagaaccggt catggccgca ataaaatatc
4921 tttattttca ttacatctgt gtgttggttt tttgtgtgtt cgaactagat gctgtcgacc
4981 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
5041 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
5101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
5161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
5221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
5281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
5341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
5401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
5461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
5521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
5581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
5641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
5701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
5761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
5821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
5881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
5941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
6001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagcggccgc aaatgctaaa
6061 ccactgcagt gg
//
