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PPRE X3-TK-luc

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PPRE X3-TK-luc載體質粒基本信息

質粒名稱: PPRE X3-TK-luc 
出品公司: 愛迪基因
目錄編號: 1015
存儲實驗室: Bruce Spiegelman

載體骨架基本信息
載體名稱: ptkLUC (modified?)
載體出品公司: --
載體抗性: 氨芐青霉素
空載體大小: --
完整質粒大小: --
載體修飾: --
載體類型: 哺乳動物表達,熒光素酶
篩選標記: --
高拷貝/低拷貝: 高拷貝
生長條件: 37 ℃

插入基因信息
插入基因名稱: PPRE,PPRE-luc,DR1X3,PPAR response element
物種來源: M. musculus (mouse)
插入大小: 100 bp
基因突變: --
GenBank編號 --
啟動子 --
標簽/融合蛋白: --
克隆方法: 限制性內切酶
5' 克隆位點: --
3' 克隆位點: --
5' 測序引物 M13pUC-fwd
3' 測序引物 LucNrev

文獻引用方法
材料和方法部分: PPRE X3-TK-luc was a gift from Bruce Spiegelman (Addgene plasmid # 1015 ; //n2t.net/addgene:1015 ; RRID:Addgene_1015)
參考文獻部分: ADD1/SREBP1 activates PPARgamma through the production of endogenous ligand. Kim JB, Wright HM, Wright M, Spiegelman BM. Proc Natl Acad Sci U S A 1998 Apr 14;95(8):4333-7. 10.1073/pnas.95.8.4333 PubMed 9539737

PPRE X3-TK-luc質粒圖譜載體圖譜和PPRE X3-TK-luc載體序列質粒序列多克隆位點信息

PPRE X3-TK-luc質粒圖譜

PPRE X3-TK-luc質粒載體簡介

PPRE X3-TK-luc質粒序列載體序列

LOCUS       PPRE X3-TK-luc             6763 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  PPAR reporter.  3xDR1 sites upstream of a luciferase reporter..
ACCESSION   .
VERSION     .
KEYWORDS    PPRE X3-TK-luc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6763)
  AUTHORS   Kim JB, Wright HM, Wright M, Spiegelman BM
  TITLE     ADD1/SREBP1 activates PPARgamma through the production of endogenous
            ligand.
  JOURNAL   Proc Natl Acad Sci U S A 1998 Apr 14;95(8):4333-7.
  PUBMED    9539737
REFERENCE   2  (bases 1 to 6763)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6763
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..63
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             123..1775
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(189..207)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          1906..1971
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2101..2121
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2546..2680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2583..2602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2637..2656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2698..2714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2698..2714)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2711..2733)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2722..2738
                     /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(2746..2776)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2791..2812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2929..2946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3100..3688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3180..3199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3859..4719)
                     /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     4482..4501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4720..4824)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4892..4910
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4948..4970)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5070..5089
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5571..6084)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5766..5787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5978..5997
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6510..6532
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6524..6541
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6525..6541
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca gcgacccgct
       61 taacagcgtc aacagcgtgc cgcagatctc gaggagcttg gcattccggt actgttggta
      121 aaatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatcct ctagaggatg
      181 gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa
      241 ttgcttttac agatgcacat atcgaggtga acatcacgta cgcggaatac ttcgaaatgt
      301 ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg
      361 tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag
      421 ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgaaca
      481 tttcgcagcc taccgtagtg tttgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc
      541 aaaaaaaatt accaataatc cagaaaatta ttatcatgga ttctaaaacg gattaccagg
      601 gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg
      661 attttgtacc agagtccttt gatcgtgaca aaacaattgc actgataatg aattcctctg
      721 gatctactgg gttacctaag ggtgtggccc ttccgcatag aactgcctgc gtcagattct
      781 cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg
      841 ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat
      901 ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tttacgatcc cttcaggatt
      961 acaaaattca aagtgcgttg ctagtaccaa ccctattttc attcttcgcc aaaagcactc
     1021 tgattgacaa atacgattta tctaatttac acgaaattgc ttctgggggc gcacctcttt
     1081 cgaaagaagt cggggaagcg gttgcaaaac gcttccatct tccagggata cgacaaggat
     1141 atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg
     1201 gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga
     1261 aaacgctggg cgttaatcag agaggcgaat tatgtgtcag aggacctatg attatgtccg
     1321 gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt
     1381 ctggagacat agcttactgg gacgaagacg aacacttctt catagttgac cgcttgaagt
     1441 ctttaattaa atacaaagga tatcaggtgg cccccgctga attggaatcg atattgttac
     1501 aacaccccaa catcttcgac gcgggcgtgg caggtcttcc cgacgatgac gccggtgaac
     1561 ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg
     1621 attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg
     1681 acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca
     1741 taaaggccaa gaagggcgga aagtccaaat tgtaaaatgt aactgtattc agcgatgacg
     1801 aaattcttag ctattgtaat actctagagg atctttgtga aggaacctta cttctgtggt
     1861 gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt
     1921 ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc
     1981 tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc
     2041 tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct
     2101 ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt
     2161 ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag
     2221 gaaaaagctg cactgctata caagaaaatt atggaaaaat attctgtaac ctttataagt
     2281 aggcataaca gttataatca taacatactg ttttttctta ctccacacag gcatagagtg
     2341 tctgctatta ataactatgc tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg
     2401 gttaataagg aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac
     2461 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2521 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2641 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gggaattcgt aatcatggtc
     2701 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2761 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2821 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2881 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2941 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3001 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3481 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3541 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3661 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3721 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3781 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3841 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3901 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3961 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4021 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4081 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4141 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4201 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4261 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4321 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4381 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4441 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4501 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4561 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4621 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4681 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4741 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4801 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     4861 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     4921 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4981 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5041 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5101 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgatt
     5161 ccaacatcca ataaatcata caggcaaggc aaagaattag caaaattaag caataaagcc
     5221 tcagagcata aagctaaatc ggttgtacca aaaacattat gaccctgtaa tacttttgcg
     5281 ggagaagcct ttatttcaac gcaaggataa aaatttttag aaccctcata tattttaaat
     5341 gcaatgcctg agtaatgtgt aggtaaagat tcaaacgggt gagaaaggcc ggagacagtc
     5401 aaatcaccat caatatgata ttcaaccgtt ctagctgata aattcatgcc ggagagggta
     5461 gctatttttg agaggtctct acaaaggcta tcaggtcatt gcctgagagt ctggagcaaa
     5521 caagagaatc gatgaacggt aatcgtaaaa ctagcatgtc aatcatatgt accccggttg
     5581 ataatcagaa aagccccaaa aacaggaaga ttgtataagc aaatatttaa attgtaagcg
     5641 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     5701 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     5761 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     5821 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     5881 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     5941 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6001 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6061 ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgagcacgta taacgtgctt
     6121 tcctcgttag aatcagagcg ggagctaaac aggaggccga ttaaagggat tttagacagg
     6181 aacggtacgc cagaatcctg agaagtgttt ttataatcag tgaggccacc gagtaaaaga
     6241 gtctgtccat cacgcaaatt aaccgttgtc gcaatacttc tttgattagt aataacatca
     6301 cttgcctgag tagaagaact caaactatcg gccttgctgg taatatccag aacaatatta
     6361 ccgccagcca ttgcaacgga atcgccattc gccattcagg ctgcgcaact gttgggaagg
     6421 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6481 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     6541 tgccaagctt gcatgcctgc aggtcgacag gggaccagga caaaggtcac gttcgggagt
     6601 cgacagggga ccaggacaaa ggtcacgttc gggagtcgac aggggaccag gacaaaggtc
     6661 acgttcggga gtcgactcta gaggatccgg ccccgcccag cgtcttgtca ttggcgaatt
     6721 cgaacacgca gatgcagtcg gggcggcgcg gtccgaggtc cac
//

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