| 质粒名称: | pcDNA flag PPAR gamma |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 8895 |
| 存储实验室: | Bruce Spiegelman |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 myc-His A |
| 载体出品公司: | Invitrogen |
| 空载体大小: | 5500bp |
| 完整质粒大小: | 7042 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | PPAR gamma、PPARg |
| 物种来源: | PPARg |
| 插入大小: | 1547bp |
| GenBank编号 | NM_011146 |
| 启动子: | CMV |
| 标签/融合蛋白: | flag (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NotI (not destroyed) |
| 3’克隆位点: | XbaI (not destroyed) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA flag PPAR gamma was a gift from Bruce Spiegelman (Addgene plasmid # 8895 ; http://n2t.net/addgene:8895 ; RRID:Addgene_8895) |
| 参考文献部分: | Degradation of the peroxisome proliferator-activated receptor gamma is linked to ligand-dependent activation. Hauser S, Adelmant G, Sarraf P, Wright HM, Mueller E, Spiegelman BM. J Biol Chem. 2000 Jun 16. 275(24):18527-33. 10.1074/jbc.M001297200 PubMed 10748014 |
买家导航
pcDNA flag PPAR gamma载体质粒基本信息
pcDNA flag PPAR gamma质粒图谱载体图谱和pcDNA flag PPAR gamma载体序列质粒序列多克隆位点信息
pcDNA flag PPAR gamma质粒载体简介
pcDNA flag PPAR gamma质粒序列载体序列
LOCUS pcDNA flag PPAR gamma 7042 bp DNA circular SYN 14-APR-2025
DEFINITION Mammalian expression of flag-tagged PPAR gamma.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7042)
AUTHORS Hauser S, Adelmant G, Sarraf P, Wright HM, Mueller E, Spiegelman BM
TITLE Degradation of the peroxisome proliferator-activated receptor gamma
is linked to ligand-dependent activation.
JOURNAL J Biol Chem. 2000 Jun 16. 275(24):18527-33.
PUBMED 10748014
REFERENCE 2 (bases 1 to 7042)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem. 2000 Jun 16. 275(24):18527-33."
COMMENT Sequence Label: pcDNA flag PPAR gamma
FEATURES Location/Qualifiers
source 1..7042
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(46..65)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 237..616
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 617..820
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 771..791
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 865..884
/label=T7
/note="T7 promoter, forward primer"
promoter 865..883
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 988..1011
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2548..2577
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2593..2610
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2633..2650)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2639..2863
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2909..3337
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2996..3015)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3206..3227
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3346..3366)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3351..3680
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3531..3666
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3593..3612
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3747..4541
/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(3801..3820)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4411..4430
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4717..4838
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4754..4773)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4808..4827
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4887..4903)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4887..4903)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4900..4922)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4911..4927
/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(join(4935..4941,4942..4959,4960..4965))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4980..5001
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5118..5135)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5289..5877)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5369..5388)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(6048..6839,6840..6908))
/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 6671..6690
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6909..7013)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
901 gttaagcttg gtaccgagct cggatccact agtccagtgt ggtggaattc tgcagatatc
961 cagcacagtg gcggccgcac caccatggac tacaaagacg atgacgataa agcaatgggt
1021 gaaactctgg gagattctcc tgttgaccca gagcatggtg ccttcgctga tgcactgcct
1081 atgagcactt cacaagaaat taccatggtt gacacagaga tgccattctg gcccaccaac
1141 ttcggaatca gctctgtgga cctctccgtg atggaagacc actcgcattc ctttgacatc
1201 aagcccttta ccacagttga tttctccagc atttctgctc cacactatga agacattcca
1261 ttcacaagag ctgacccaat ggttgctgat tacaaatatg acctgaagct ccaagaatac
1321 caaagtgcga tcaaagtaga acctgcatct ccaccttatt attctgaaaa gacccagctc
1381 tacaacaggc ctcatgaaga accttctaac tccctcatgg ccattgagtg ccgagtctgt
1441 ggggataaag catcaggctt ccactatgga gttcatgctt gtgaaggatg caagggtttt
1501 ttccgaagaa ccatccgatt gaagcttatt tatgataggt gtgatcttaa ctgccggatc
1561 cacaaaaaaa gtagaaataa atgtcagtac tgtcggtttc agaagtgcct tgctgtgggg
1621 atgtctcaca atgccatcag gtttgggcgg atgccacagg ccgagaagga gaagctgttg
1681 gcggagatct ccagtgatat cgaccagctg aacccagagt ctgctgatct gcgagccctg
1741 gcaaagcatt tgtatgactc atacataaag tccttcccgc tgaccaaagc caaggcgagg
1801 gcgatcttga caggaaagac aacggacaaa tcaccatttg tcatctacga catgaattcc
1861 ttaatgatgg gagaagataa aatcaagttc aaacatatca cccccctgca ggagcagagc
1921 aaagaggtgg ccatccgaat ttttcaaggg tgccagtttc gatccgtaga agccgtgcaa
1981 gagatcacag agtatgccaa aaatatccct ggtttcatta accttgattt gaatgaccaa
2041 gtgactctgc tcaagtatgg tgtccatgag atcatctaca cgatgctggc ctccctgatg
2101 aataaagatg gagtcctcat ctcagagggc caaggattca tgaccaggga gttcctcaaa
2161 agcctgcgga agccctttgg tgactttatg gagcctaagt ttgagtttgc tgtgaagttc
2221 aatgcactgg aattagatga cagtgacttg gctatattta tagctgtcat tattctcagt
2281 ggagaccgcc caggcttgct gaacgtgaag cccatcgagg acatccaaga caacctgctg
2341 caggccctgg aactgcagct caagctgaat cacccagagt cctctcagct gttcgccaag
2401 gtgctccaga agatgacaga cctcaggcag atcgtcacag agcacgtgca gctactgcat
2461 gtgatcaaga agacagagac agacatgagc cttcaccccc tgctccagga gatctacaag
2521 gacttgtatt agtctagagg gcccttcgaa caaaaactca tctcagaaga ggatctgaat
2581 atgcataccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
2641 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
2701 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
2761 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
2821 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
2881 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
2941 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
3001 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
3061 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
3121 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
3181 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
3241 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
3301 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
3361 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
3421 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
3481 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
3541 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
3601 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
3661 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
3721 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
3781 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
3841 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
3901 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
3961 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
4021 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
4081 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
4141 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
4201 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
4261 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
4321 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
4381 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
4441 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
4501 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
4561 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
4621 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
4681 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
4741 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
4801 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
4861 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
4921 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
4981 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
5041 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
5101 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
5161 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
5221 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
5281 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
5341 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
5401 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
5461 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
5521 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
5581 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
5641 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
5701 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
5761 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
5821 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
5881 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
5941 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
6001 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
6061 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
6121 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
6181 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
6241 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
6301 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
6361 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
6421 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
6481 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
6541 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
6601 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
6661 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
6721 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
6781 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
6841 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
6901 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
6961 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
7021 ccccgaaaag tgccacctga cg
//
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