| 质粒名称: | PPRE X3-TK-luc |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 1015 |
| 存储实验室: | Bruce Spiegelman |
|
载体骨架基本信息 |
|
| 载体名称: | ptkLUC (modified?) |
| 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: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，荧光素酶 |
| 筛选标记: | -- |
| 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: | PPRE,PPRE-luc,DR1X3,PPAR response element |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 100 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: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | M13pUC-fwd |
| 3' 测序引物 | LucNrev |
|
文献引用方法 |
|
| 材料和方法部分: | PPRE X3-TK-luc was a gift from Bruce Spiegelman (Addgene plasmid # 1015 ; http://n2t.net/addgene:1015 ; RRID:Addgene_1015) |
| 参考文献部分: | 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 |
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 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
http://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
//
