| 质粒名称: | pGL4-UPRE-luc2P-Hygro |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 101788 |
| 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: | 6078 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 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: | 3xUPRE |
| 物种来源: | Synthetic |
| 插入大小: | 87 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-UPRE-luc2P-Hygro was a gift from Seiichi Oyadomari (Addgene plasmid # 101788 ; http://n2t.net/addgene:101788 ; RRID:Addgene_101788) |
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-UPRE-luc2P-Hygro 6078 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Luciferase reporter plasmid containing three tandem repeats of the
unfolded protein response element (UPRE).
ACCESSION .
VERSION .
KEYWORDS pGL4-UPRE-luc2P-Hygro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6078)
AUTHORS Seiichi Oyadomari
TITLE Oyadomari Lab Luciferase reporters
JOURNAL unpublished
REFERENCE 2 (bases 1 to 6078)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6078
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 145..176
/label=minP
/note="minimal TATA-box promoter with low basal activity"
regulatory 203..212
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 209..1858
/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 1862..1981
/codon_start=1
/product="PEST degradation sequence from mouse ornithine
decarboxylase"
/label=hPEST
/note="human codon-optimized"
/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
polyA_signal 2033..2154
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2044..2063)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2098..2117
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 2348..2705
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2371..2391)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2556..2691
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2618..2637
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2736..3773
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
EVGRV"
RBS 3752..3760
/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 3797..3845
/note="synthetic polyadenylation signal"
primer_bind complement(4001..4018)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4172..4760)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4252..4271)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4960..5820)
/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 5925..5973
/note="synthetic polyadenylation signal"
misc_feature 5987..6078
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 6027..6046
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 ggcctaactg gccggtacct gagctcgcta gcctcgagac aggtgctgac gtggcattcc
61 tcgagacagg tgctgacgtg gcattcctcg agacaggtgc tgacgtggca ttcagatctg
121 gcctcggcgg ccaagcttag acactagagg gtatataatg gaagctcgac ttccagcttg
181 gcaatccggt actgttggta aagccaccat ggaagatgcc aaaaacatta agaagggccc
241 agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca aagccatgaa
301 gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg aggtggacat
361 tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga agcgctatgg
421 gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc
481 cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca tctacaacga
541 gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa
601 agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa agatcatcat
661 catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg tgacttccca
721 tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc gggacaaaac
781 catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg tagccctacc
841 gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg gcaaccagat
901 catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg gcatgttcac
961 cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga
1021 gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg tgcccacact
1081 atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca acttgcacga
1141 gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt
1201 ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg ccattctgat
1261 cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct tcttcgaggc
1321 taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg
1381 cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta caaacgctct
1441 catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg aggacgagca
1501 cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc aggtagcccc
1561 agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg
1621 cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa
1681 aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa ccgccaagaa
1741 gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga
1801 cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga tcgccgtgaa
1861 ttctcacggc ttccctcccg aggtggagga gcaggccgcc ggcaccctgc ccatgagctg
1921 cgcccaggag agcggcatgg atagacaccc tgctgcttgc gccagcgcca ggatcaacgt
1981 ctaaggccgc gactctagag tcggggcggc cggccgcttc gagcagacat gataagatac
2041 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
2101 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
2161 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
2221 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc
2281 ttccgctgat ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
2341 ctaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
2401 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
2461 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
2521 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
2581 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
2641 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
2701 gcaaaaagct cgattcttct gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta
2761 ccagcgttga aaaatttctc atcgagaagt tcgacagtgt gagcgacctg atgcagttgt
2821 cggagggcga agagagccga gccttcagct tcgatgtcgg cggacgcggc tatgtactgc
2881 gggtgaatag ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca
2941 gcgctgcact acccatcccc gaagtgttgg acatcggcga gttcagcgag agcctgacat
3001 actgcatcag tagacgcgcc caaggcgtta ctctccaaga cctccccgaa acagagctgc
3061 ctgctgtgtt acagcctgtc gccgaagcta tggatgctat tgccgccgcc gacctcagtc
3121 aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg ccagtacaca acctggcggg
3181 atttcatttg cgccattgct gatccccatg tctaccactg gcagaccgtg atggacgaca
3241 ccgtgtccgc cagcgtagct caagccctgg acgaactgat gctgtgggcc gaagactgtc
3301 ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa caacgtcctg accgacaacg
3361 gccgcatcac cgccgtaatc gactggtccg aagctatgtt cggggacagt cagtacgagg
3421 tggccaacat cttcttctgg cggccctggc tggcttgcat ggagcagcag actcgctact
3481 tcgagcgccg gcatcccgag ctggccggca gccctcgtct gcgagcctac atgctgcgca
3541 tcggcctgga tcagctctac cagagcctcg tggacggcaa cttcgacgat gctgcctggg
3601 ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt cgcacacaaa
3661 tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg gccgacagcg
3721 gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt taaactctag
3781 aaccggtcat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
3841 gtgtgttcga actagatgct gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
3901 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
3961 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
4021 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
4081 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
4141 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
4201 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
4261 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
4321 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
4381 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
4441 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
4501 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
4561 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
4621 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
4681 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
4741 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
4801 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
4861 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
4921 ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga
4981 ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt
5041 gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg
5101 agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga
5161 gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga
5221 tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg
5281 catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc
5341 aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc
5401 gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca
5461 caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac
5521 caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg
5581 ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc
5641 ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct
5701 tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac
5761 aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat
5821 actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg tctctcaagg
5881 ataagtaagt aatattaagg tacgggaggt attggacagg ccgcaataaa atatctttat
5941 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc
6001 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
6061 gtgccagaac atttctct
//
