| 质粒名称: | TFEB promoter-luciferase reporter |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 66801 |
| 存储实验室: | Albert La Spada |
|
载体骨架基本信息 |
|
| 载体名称: | pGL3-Basic |
| 载体出品公司: | Promega |
| 载体抗性: | 氨苄青霉素 |
| 载体类型: | 荧光素酶表达 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | TFEB promoter,Tcfeb,bHLHe35 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 2000 bp |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Nhei（销毁后未知） |
| 3' 克隆位点: | HindIII（销毁后未知） |
| 5' 测序引物 | RVprimer3 (CTAGCAAAATAGGCTGTCCC) |
| 3' 测序引物 | EBV rev |
|
文献引用方法 |
|
| 材料和方法部分: | TFEB promoter-luciferase reporter was a gift from Albert La Spada (Addgene plasmid # 66801 ; http://n2t.net/addgene:66801 ; RRID:Addgene_66801) |
| 参考文献部分: | PGC-1alpha rescues Huntington's disease proteotoxicity by preventing oxidative stress and promoting TFEB function. Tsunemi T, Ashe TD, Morrison BE, Soriano KR, Au J, Roque RA, Lazarowski ER, Damian VA, Masliah E, La Spada AR. Sci Transl Med. 2012 Jul 11;4(142):142ra97. doi: 10.1126/scitranslmed.3003799. 10.1126/scitranslmed.3003799 PubMed 22786682 |
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 TFEB promoter-luciferase reporter 7001 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Reporter for activity of TFEB gene promoter.
ACCESSION .
VERSION .
KEYWORDS TFEB promoter-luciferase reporter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7001)
AUTHORS Tsunemi T, Ashe TD, Morrison BE, Soriano KR, Au J, Roque RA,
Lazarowski ER, Damian VA, Masliah E, La Spada AR
TITLE PGC-1alpha rescues Huntington's disease proteotoxicity by preventing
oxidative stress and promoting TFEB function.
JOURNAL Sci Transl Med. 2012 Jul 11;4(142):142ra97. doi:
10.1126/scitranslmed.3003799.
PUBMED 22786682
REFERENCE 2 (bases 1 to 7001)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7001
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 69..117
/note="synthetic polyadenylation signal"
misc_feature 131..222
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 171..190
/label=RVprimer3
/note="pGL3 vector, forward primer"
regulatory 2494..2503
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2500..4152
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(2566..2584)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
polyA_signal 4193..4314
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4204..4223)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4258..4277
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(4562..4579)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4733..5321)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4813..4832)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5492..6352)
/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 6115..6134
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6353..6457)
/gene="bla"
/label=AmpR promoter
rep_origin 6484..6939
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6571..6590)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6781..6802
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 ttacgccagc ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga
61 gcggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
121 cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat
181 aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct
241 cttacgcgtg ctagtgtggc cagatagggt gtggtgtgga gacagggaca tcactctgta
301 ggagctagag aaggggtcgt gggaggtggt ggaggctgca gctggtagct gctacaggat
361 gggggaaggg taggttgtca ttgcctgggt tgttgggggc ttccctgtag agcatgggct
421 gctcaggccc cttcactggg cagagcatga gagaagatgc agtgctggcc tctgcatgca
481 gcaggacaca caggacagtg ggtagtctca ctctattaca gagctgctta gatagtggct
541 ggagtcttga ttgcagatat gggacctagg ctccactcct ggttctggat caaccagttt
601 cccaactggg aacaactctg ccccccaggc ttccccttcc cagcatgggg tagaccagat
661 gtcccagccc ccaggcctcc cttccccagc atggggtaga ccagatacct ggggtggggt
721 cctagctttg tcctgcactg tgcctctgat ttcccttctt tatgaaagac tgtggtttgt
781 aattaagcaa gggcctccaa acagggctga tggaggcctt ggatgccggg tttgaagacg
841 ctagctagat tgtaccagtc tcatgtactc agactgccag gcactgcaca aagctgccag
901 tttttactaa atgcagagga agataaactg aggctccagg attatagctt gaccaaggtc
961 acataacaaa gaatcagcga gcgctctgct ccaccagccc tttggggaag ccgttgctca
1021 atcatgaagt catgccttgc caaatccctc tagtccctaa ctgtactgtc atggtctcag
1081 atacggccag accagtgcct atcagtcttg ggctgtccct gagaagtcac tgggttctgc
1141 tctttccttt gtggcctaga gaggggaagg gacttggtta ctgtcactca ttgagctaga
1201 gacagagcga gcatgccgag ttgcctgccc cccttcagcc ttggttgggg cctcagccac
1261 accttcccac agtgggtcaa gctctgagct ccgagcagtg aggagccctg aggtggtgat
1321 tctctgcctg tgggtcttcc tttctctcgc cgactttggg cgccaggagc cagggacggg
1381 gtacatctcc agtgcctggt tcatggataa tatccattct catttgcttc atgcaatggc
1441 ttctttctgt ctcccatgac ccaacggcta gaggccgaag gtctagcagt aaacaaacag
1501 atgaaatgtg tcccctcagg ggacttgcat tcctagtaag ggacagacat ggaataacaa
1561 taacaaacgt aataagaagc aaaaaaaaaa aatatgttct gtcctagatg gtggcatctg
1621 atatagagaa gcaaggtggt ggtggtatgc cattttagca cagtggtcct gaaatggcac
1681 tctgaggtga cattagagtc tctcggagga ggtgaagaag tgagccttga tggtatctgt
1741 gtaaagaaca ttccaggtgg aggcaacagc aagtgcaaag gtgggggggg gggacggtgg
1801 ggggcggaga aagcatgcct ggagtgtttg aaggatagtt taaggctggg ggtgtggctg
1861 gagcagggag aacaagggag agaagcatgt gttgggggcg ggaagtctat gggggaccat
1921 gagggtgcag cagattacag aggaccattt aggctgggtt taaggattta ggcttttgtt
1981 ccgatgaact gggagctatg gagagttggg agcagacagc agggcaggaa ctaacaggat
2041 gatggagcta ctgtgagcag atggtggggg aggagcacgg aggagcatct ggaggctcct
2101 caggccctga ggaggcgtgg tgcaggctgg cctggggagg ggcgggagag ggggccggag
2161 ctgaggccag gtatgaaata ctcaggatgt ggatgtgaca gcgaggggtc aggtggactc
2221 ctgcagtcac tgaaggacag tgttcacctc tggtctagaa ctcagggaac ttgggttcat
2281 ggctcagctc gctcagtggt cttgggcaaa tcccttctgc ccggactcag tttctccttg
2341 tgcacaatgg gagcaaccat acttatgcca gcctgtgctt aaagactacg aagatgatga
2401 atacttcatg ggcctgtctc ccctcgacta cagggagccc gaaccaacag ctgccatccc
2461 ggggaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
2521 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
2581 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
2641 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
2701 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
2761 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
2821 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
2881 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
2941 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
3001 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
3061 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
3121 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
3181 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
3241 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
3301 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
3361 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
3421 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
3481 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
3541 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
3601 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
3661 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
3721 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
3781 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
3841 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
3901 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
3961 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
4021 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
4081 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
4141 atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
4201 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
4261 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
4321 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
4381 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
4441 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
4501 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
4561 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
4621 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
4681 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
4741 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
4801 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
4861 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
4921 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
4981 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
5041 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
5101 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
5161 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
5221 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
5281 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
5341 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
5401 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
5461 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
5521 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
5581 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
5641 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
5701 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
5761 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
5821 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
5881 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
5941 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
6001 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
6061 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
6121 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
6181 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
6241 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
6301 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
6361 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
6421 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
6481 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
6541 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
6601 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
6661 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
6721 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
6781 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
6841 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
6901 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
6961 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct a
//
