| 质粒名称: | PPRE-H2B-eGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 84393 |
| 存储实验室: | Severine Degrelle, Thierry Fournier |
|
载体骨架基本信息 |
|
| 载体名称: | H2B-GFP |
| 载体出品公司: | Clontech |
| 空载体大小: | 5100bp |
| 完整质粒大小: | 6000bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 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: | PPARG response element、PPRE、ApoCIII promotor |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1200bp |
| 启动子: | PPRE |
| 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: | AseI (not destroyed) |
| 3’克隆位点: | NheI (not destroyed) |
| 5’测序引物 | PPRE-AseI |
| 3’测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | PPRE-H2B-eGFP was a gift from Severine Degrelle & Thierry Fournier (Addgene plasmid # 84393 ; http://n2t.net/addgene:84393 ; RRID:Addgene_84393) |
| 参考文献部分: | New Transcriptional Reporters to Quantify and Monitor PPARgamma Activity. Degrelle SA, Shoaito H, Fournier T. PPAR Res. 2017;2017:6139107. doi: 10.1155/2017/6139107. Epub 2017 Nov 1. 10.1155/2017/6139107 PubMed 29225614 |
买家导航
PPRE-H2B-eGFP载体质粒基本信息
PPRE-H2B-eGFP质粒图谱载体图谱和PPRE-H2B-eGFP载体序列质粒序列多克隆位点信息
PPRE-H2B-eGFP质粒载体简介
PPRE-H2B-eGFP质粒序列载体序列
LOCUS PPRE-H2B-eGFP 5664 bp DNA circular SYN 25-AUG-2024
DEFINITION Fluorescent reporter for PPARgamma activity.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5664)
AUTHORS Degrelle SA, Shoaito H, Fournier T
TITLE New Transcriptional Reporters to Quantify and Monitor PPARgamma
Activity.
JOURNAL PPAR Res. 2017;2017:6139107. doi: 10.1155/2017/6139107. Epub 2017
Nov 1.
PUBMED 29225614
REFERENCE 2 (bases 1 to 5664)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PPAR Res.";
date: "2017"; volume: "2017:6139107. doi"; pages: "
10.1155/2017/6139107. Epub 2017 Nov 1"
FEATURES Location/Qualifiers
source 1..5664
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1302..1679
/codon_start=1
/gene="HIST1H2BJ"
/product="human histone H2B"
/label=H2B
/translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
GELAKHAVSEGTKAITKYTSAK"
regulatory 1692..1701
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1698..2417
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1698..2414
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1743..1764)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2004..2023)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2351..2372
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2540..2661
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2577..2596)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2631..2650
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2668..3123)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2805..2826)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3017..3036
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3150..3254
/gene="bla"
/label=AmpR promoter
promoter 3256..3613
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 3256..3447
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(3279..3299)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3464..3599
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3648..4442
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 3648..4442
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3702..3721)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4312..4331
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4630..4649)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4674..4721
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5050..5638
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5539..5558
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
61 tgtggataac cgtattaccg ccatgcatta gttattaata atgcctgcag gtcaattctg
121 agggcagagc cgggccactt tctcaggcct ctgatttcat actgtggtgt tagttacttc
181 tgagaggaca gcttgctgcc agagctctat tttttatgtt agaggctcct tctgcctgca
241 gactctgctg tctgggaagg gcacagcgtt aggagggaga gggaggtgtg agtccctccg
301 tggacccgct gctttgtact tctctatctc atttcctttt cagcaccact ctgggaaatc
361 agtattccag ccccatttta tcctcagaaa attgaggctc tgagatgtta tctctgtgac
421 ctgggtccta ttacgtgcca aaggcatcat ttaagcctaa gatgtcctgg ctccaaggtg
481 tcagcatctg gaagacaggc gccctcatcc tgccatccct gctgcggctt cactgtgggc
541 ccaggggaca tctcagcccc gagaagggtc agcggcccct cctggaccac cgactccccg
601 cagaactcct ctgtgccctc tcctcaccag accttgttcc tcccagttgc tcccacagcc
661 agggggcagt gagggctgct cttcccccag ccccactgag gaacccagga aggtgaacga
721 gagaatcagt cctggtgggg gctggggagg gccccagaca tgagaccagc tcctccccca
781 ggggatgtta tcagtgggtc cagagggcaa aatagggagc ctggtggagg gaggggcctg
841 aaattccagt gtgaaaggct gagatgggcc cgaggcccct ggcctatgtc caagccattt
901 cccctctcac cagcctctcc ctggggagcc agtcagctag gaaggaatga gggctcccca
961 ggcccacccc agttcctgag ctcatctggg ctgcagggct ggcgggacag cagcgtggac
1021 tcagtctcct agggatttcc caactctccc gcccgcttgc tgcatctgga caccctgcct
1081 caggccctca tctccactgg tcagcaggtg acctttgccc agcgccctgg gtcctcagtg
1141 cctgctgccc tggagatgat ataaaacagg tcagaaccct cctgcctgtc tgctcagttc
1201 atccctagag gcaggactct agaggatctg cgatcgctag cgctaccgga ctcagatctc
1261 gagctcaagc ttcgaattct gcagtcgacg gtaccgccac catgccagag ccagcgaagt
1321 ctgctcccgc cccgaaaaag ggctccaaga aggcggtgac taaggcgcag aagaaaggcg
1381 gcaagaagcg caagcgcagc cgcaaggaga gctattccat ctatgtgtac aaggttctga
1441 agcaggtcca ccctgacacc ggcatttcgt ccaaggccat gggcatcatg aattcgtttg
1501 tgaacgacat tttcgagcgc atcgcaggtg aggcttcccg cctggcgcat tacaacaagc
1561 gctcgaccat cacctccagg gagatccaga cggccgtgcg cctgctgctg cctggggagt
1621 tggccaagca cgccgtgtcc gagggtacta aggccatcac caagtacacc agcgctaagg
1681 atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
1741 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
1801 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
1861 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
1921 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
1981 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
2041 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
2101 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
2161 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
2221 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
2281 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
2341 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
2401 acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
2461 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
2521 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
2581 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
2641 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
2701 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
2761 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
2821 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
2881 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
2941 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
3001 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
3061 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
3121 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
3181 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
3241 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
3301 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
3361 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
3421 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
3481 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
3541 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
3601 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
3661 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
3721 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
3781 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
3841 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
3901 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
3961 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
4021 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
4081 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
4141 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
4201 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
4261 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
4321 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
4381 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
4441 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
4501 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
4561 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
4621 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
4681 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
4741 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
4801 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
4861 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
4921 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
4981 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
5041 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
5101 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
5161 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
5221 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
5281 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
5341 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
5401 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
5461 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
5521 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
5581 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
5641 cgccagcaac gcggcctttt tacg
//
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