| 质粒名称: | pGL4-AARE-luc2P-Hygro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101787 |
| 存储实验室: | Seiichi Oyadomari |
|
载体骨架基本信息 |
|
| 载体名称: | pGL4.29 |
| 载体出品公司: | Promega |
| 完整质粒大小: | 6054bp |
| 载体类型: | 哺乳细胞表达\Luciferase |
| 筛选标记: | Hygromycin |
| 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: | 3xAARE |
| 物种来源: | Synthetic |
| 插入大小: | 63 bp |
| 启动子: | minimal TATA-box promoter with low basal activity |
| 标签/融合蛋白: | Luciferase (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NheI (未破坏) |
| 3’克隆位点: | BglII (未破坏) |
| 5’测序引物 | ctaactggccggtacctgag |
| 3’测序引物 | aacagtaccggattgccaag |
|
文献引用方法 |
|
| 材料和方法部分: |
pGL4-AARE-luc2P-Hygro was a gift from Seiichi Oyadomari (Addgene plasmid # 101787 ; http://n2t.net/addgene:101787 ; RRID:Addgene_101787)
|
| 参考文献部分: | Skeletal muscle-specific eukaryotic translation initiation factor 2alpha phosphorylation controls amino acid metabolism and fibroblast growth factor 21-mediated non-cell-autonomous energy metabolism. Miyake M, Nomura A, Ogura A, Takehana K, Kitahara Y, Takahara K, Tsugawa K, Miyamoto C, Miura N, Sato R, Kurahashi K, Harding HP, Oyadomari M, Ron D, Oyadomari S. FASEB J. 2016 Feb;30(2):798-812. doi: 10.1096/fj.15-275990. Epub 2015 Oct 20. 10.1096/fj.15-275990 PubMed 26487695 |
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 pGL4-AARE-luc2P-Hygro 6054 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Luferase reporter plasmid containing three tandem repeats of the
amino acid response element (AARE).
ACCESSION .
VERSION .
KEYWORDS pGL4-AARE-luc2P-Hygro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6054)
AUTHORS Miyake M, Nomura A, Ogura A, Takehana K, Kitahara Y, Takahara K,
Tsugawa K, Miyamoto C, Miura N, Sato R, Kurahashi K, Harding HP,
Oyadomari M, Ron D, Oyadomari S
TITLE Skeletal muscle-specific eukaryotic translation initiation factor
2alpha phosphorylation controls amino acid metabolism and fibroblast
growth factor 21-mediated non-cell-autonomous energy metabolism.
JOURNAL FASEB J. 2016 Feb;30(2):798-812. doi: 10.1096/fj.15-275990. Epub
2015 Oct 20.
PUBMED 26487695
REFERENCE 2 (bases 1 to 6054)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6054
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 121..152
/label=minP
/note="minimal TATA-box promoter with low basal activity"
regulatory 179..188
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 185..1834
/codon_start=1
/product="firefly luciferase"
/label=luciferase
/note="synthetic luc2 version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
CDS 1838..1957
/codon_start=1
/product="PEST degradation sequence from mouse ornithine
decarboxylase"
/label=hPEST
/note="human codon-optimized"
/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
polyA_signal 2009..2130
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2020..2039)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2074..2093
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 2324..2681
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2347..2367)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2532..2667
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2594..2613
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2712..3749
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
EVGRV"
RBS 3728..3736
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
polyA_signal 3773..3821
/note="synthetic polyadenylation signal"
primer_bind complement(3977..3994)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4148..4736)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4228..4247)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4936..5796)
/codon_start=1
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
polyA_signal 5901..5949
/note="synthetic polyadenylation signal"
misc_feature 5963..6054
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 6003..6022
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 ggcctaactg gccggtacct gagctcgcta gcaacattgc atcatccccg caacattgca
61 tcatccccgc aacattgcat catccccgca gatctggcct cggcggccaa gcttagacac
121 tagagggtat ataatggaag ctcgacttcc agcttggcaa tccggtactg ttggtaaagc
181 caccatggaa gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga
241 cgggaccgcc ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac
301 catcgccttt accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat
361 gagcgttcgg ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt
421 ggtgtgcagc gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg
481 tgtggctgtg gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg
541 catcagccag cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt
601 gcaaaagaag ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca
661 gggcttccaa agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta
721 cgacttcgtg cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag
781 tggcagtacc ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt
841 cagtcatgcc cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag
901 cgtggtgcca tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg
961 ctttcgggtc gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga
1021 ctataagatt caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac
1081 tctcatcgac aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct
1141 cagcaaggag gtaggtgagg ccgtggccaa acgcttccac ctaccaggca tccgccaggg
1201 ctacggcctg acagaaacaa ccagcgccat tctgatcacc cccgaagggg acgacaagcc
1261 tggcgcagta ggcaaggtgg tgcccttctt cgaggctaag gtggtggact tggacaccgg
1321 taagacactg ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag
1381 cggctacgtt aacaaccccg aggctacaaa cgctctcatc gacaaggacg gctggctgca
1441 cagcggcgac atcgcctact gggacgagga cgagcacttc ttcatcgtgg accggctgaa
1501 gagcctgatc aaatacaagg gctaccaggt agccccagcc gaactggaga gcatcctgct
1561 gcaacacccc aacatcttcg acgccggggt cgccggcctg cccgacgacg atgccggcga
1621 gctgcccgcc gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga aggagatcgt
1681 ggactatgtg gccagccagg ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt
1741 ggacgaggtg cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc gcgagattct
1801 cattaaggcc aagaagggcg gcaagatcgc cgtgaattct cacggcttcc ctcccgaggt
1861 ggaggagcag gccgccggca ccctgcccat gagctgcgcc caggagagcg gcatggatag
1921 acaccctgct gcttgcgcca gcgccaggat caacgtctaa ggccgcgact ctagagtcgg
1981 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
2041 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
2101 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
2161 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
2221 aaaatcgata aggatccgtt tgcgtattgg gcgctcttcc gctgatctgc gcagcaccat
2281 ggcctgaaat aacctctgaa agaggaactt ggttagctac cttctgaggc ggaaagaacc
2341 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
2401 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
2461 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
2521 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
2581 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
2641 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcgat tcttctgaca
2701 ctagcgccac catgaagaag cccgaactca ccgctaccag cgttgaaaaa tttctcatcg
2761 agaagttcga cagtgtgagc gacctgatgc agttgtcgga gggcgaagag agccgagcct
2821 tcagcttcga tgtcggcgga cgcggctatg tactgcgggt gaatagctgc gctgatggct
2881 tctacaaaga ccgctacgtg taccgccact tcgccagcgc tgcactaccc atccccgaag
2941 tgttggacat cggcgagttc agcgagagcc tgacatactg catcagtaga cgcgcccaag
3001 gcgttactct ccaagacctc cccgaaacag agctgcctgc tgtgttacag cctgtcgccg
3061 aagctatgga tgctattgcc gccgccgacc tcagtcaaac cagcggcttc ggcccattcg
3121 ggccccaagg catcggccag tacacaacct ggcgggattt catttgcgcc attgctgatc
3181 cccatgtcta ccactggcag accgtgatgg acgacaccgt gtccgccagc gtagctcaag
3241 ccctggacga actgatgctg tgggccgaag actgtcccga ggtgcgccac ctcgtccatg
3301 ccgacttcgg cagcaacaac gtcctgaccg acaacggccg catcaccgcc gtaatcgact
3361 ggtccgaagc tatgttcggg gacagtcagt acgaggtggc caacatcttc ttctggcggc
3421 cctggctggc ttgcatggag cagcagactc gctacttcga gcgccggcat cccgagctgg
3481 ccggcagccc tcgtctgcga gcctacatgc tgcgcatcgg cctggatcag ctctaccaga
3541 gcctcgtgga cggcaacttc gacgatgctg cctgggctca aggccgctgc gatgccatcg
3601 tccgcagcgg ggccggcacc gtcggtcgca cacaaatcgc tcgccggagc gcagccgtat
3661 ggaccgacgg ctgcgtcgag gtgctggccg acagcggcaa ccgccggccc agtacacgac
3721 cgcgcgctaa ggaggtaggt cgagtttaaa ctctagaacc ggtcatggcc gcaataaaat
3781 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gttcgaacta gatgctgtcg
3841 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
3901 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
3961 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
4021 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
4081 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
4141 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
4201 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
4261 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
4321 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
4381 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
4441 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
4501 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
4561 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
4621 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
4681 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
4741 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
4801 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
4861 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct
4921 aaaccactgc agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct
4981 atttcgttcg tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg
5041 cttaccatca ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga
5101 tttgtcagca atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt
5161 gtccgcctcc atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt
5221 gagtagtttc cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt
5281 cggtatggct tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat
5341 attatgaaga aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc
5401 cgcggtgttg tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc
5461 cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat
5521 acggcgacca agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag
5581 tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt
5641 gccgctattg agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc
5701 ttttactttc accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa
5761 gggaatgagt gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg
5821 aagcatttat cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg
5881 ggaggtattg gacaggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
5941 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
6001 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctct
//
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