| 质粒名称: | pGL4-ERSE1-luc2P-Hygro |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 101789 |
| 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 |
| 空载体大小: | 4038 bp |
| 完整质粒大小: | 6084 bp |
| 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: | 3xERSE-I |
| 物种来源: | Synthetic |
| 插入大小: | 93 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-ERSE1-luc2P-Hygro was a gift from Seiichi Oyadomari (Addgene plasmid # 101789 ; http://n2t.net/addgene:101789 ; RRID:Addgene_101789) |
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-ERSE1-luc2P-Hygro 6084 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Luferase reporter plasmid containing three tandem repeats of the ER
stress response element-I (ERSE-I).
ACCESSION .
VERSION .
KEYWORDS pGL4-ERSE1-luc2P-Hygro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6084)
AUTHORS Seiichi Oyadomari
TITLE Oyadomari Lab Luciferase reporters
JOURNAL unpublished
REFERENCE 2 (bases 1 to 6084)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6084
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 151..182
/label=minP
/note="minimal TATA-box promoter with low basal activity"
regulatory 209..218
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 215..1864
/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 1868..1987
/codon_start=1
/product="PEST degradation sequence from mouse ornithine
decarboxylase"
/label=hPEST
/note="human codon-optimized"
/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
polyA_signal 2039..2160
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2050..2069)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2104..2123
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 2354..2711
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2377..2397)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2562..2697
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2624..2643
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2742..3779
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
EVGRV"
RBS 3758..3766
/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 3803..3851
/note="synthetic polyadenylation signal"
primer_bind complement(4007..4024)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4178..4766)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4258..4277)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4966..5826)
/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 5931..5979
/note="synthetic polyadenylation signal"
misc_feature 5993..6084
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 6033..6052
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 ggcctaactg gccggtacct gagctcgcta gcccttcacc aatcggcggc ctccacgacg
61 gccttcacca atcggcggcc tccacgacgg ccttcaccaa tcggcggcct ccacgacgga
121 gatctggcct cggcggccaa gcttagacac tagagggtat ataatggaag ctcgacttcc
181 agcttggcaa tccggtactg ttggtaaagc caccatggaa gatgccaaaa acattaagaa
241 gggcccagcg ccattctacc cactcgaaga cgggaccgcc ggcgagcagc tgcacaaagc
301 catgaagcgc tacgccctgg tgcccggcac catcgccttt accgacgcac atatcgaggt
361 ggacattacc tacgccgagt acttcgagat gagcgttcgg ctggcagaag ctatgaagcg
421 ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc gagaatagct tgcagttctt
481 catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg gccccagcta acgacatcta
541 caacgagcgc gagctgctga acagcatggg catcagccag cccaccgtcg tattcgtgag
601 caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag ctaccgatca tacaaaagat
661 catcatcatg gatagcaaga ccgactacca gggcttccaa agcatgtaca ccttcgtgac
721 ttcccatttg ccacccggct tcaacgagta cgacttcgtg cccgagagct tcgaccggga
781 caaaaccatc gccctgatca tgaacagtag tggcagtacc ggattgccca agggcgtagc
841 cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc cgcgacccca tcttcggcaa
901 ccagatcatc cccgacaccg ctatcctcag cgtggtgcca tttcaccacg gcttcggcat
961 gttcaccacg ctgggctact tgatctgcgg ctttcgggtc gtgctcatgt accgcttcga
1021 ggaggagcta ttcttgcgca gcttgcaaga ctataagatt caatctgccc tgctggtgcc
1081 cacactattt agcttcttcg ctaagagcac tctcatcgac aagtacgacc taagcaactt
1141 gcacgagatc gccagcggcg gggcgccgct cagcaaggag gtaggtgagg ccgtggccaa
1201 acgcttccac ctaccaggca tccgccaggg ctacggcctg acagaaacaa ccagcgccat
1261 tctgatcacc cccgaagggg acgacaagcc tggcgcagta ggcaaggtgg tgcccttctt
1321 cgaggctaag gtggtggact tggacaccgg taagacactg ggtgtgaacc agcgcggcga
1381 gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt aacaaccccg aggctacaaa
1441 cgctctcatc gacaaggacg gctggctgca cagcggcgac atcgcctact gggacgagga
1501 cgagcacttc ttcatcgtgg accggctgaa gagcctgatc aaatacaagg gctaccaggt
1561 agccccagcc gaactggaga gcatcctgct gcaacacccc aacatcttcg acgccggggt
1621 cgccggcctg cccgacgacg atgccggcga gctgcccgcc gcagtcgtcg tgctggaaca
1681 cggtaaaacc atgaccgaga aggagatcgt ggactatgtg gccagccagg ttacaaccgc
1741 caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg cctaaaggac tgaccggcaa
1801 gttggacgcc cgcaagatcc gcgagattct cattaaggcc aagaagggcg gcaagatcgc
1861 cgtgaattct cacggcttcc ctcccgaggt ggaggagcag gccgccggca ccctgcccat
1921 gagctgcgcc caggagagcg gcatggatag acaccctgct gcttgcgcca gcgccaggat
1981 caacgtctaa ggccgcgact ctagagtcgg ggcggccggc cgcttcgagc agacatgata
2041 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
2101 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
2161 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
2221 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtt tgcgtattgg
2281 gcgctcttcc gctgatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt
2341 ggttagctac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
2401 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
2461 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
2521 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
2581 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
2641 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
2701 gcttttgcaa aaagctcgat tcttctgaca ctagcgccac catgaagaag cccgaactca
2761 ccgctaccag cgttgaaaaa tttctcatcg agaagttcga cagtgtgagc gacctgatgc
2821 agttgtcgga gggcgaagag agccgagcct tcagcttcga tgtcggcgga cgcggctatg
2881 tactgcgggt gaatagctgc gctgatggct tctacaaaga ccgctacgtg taccgccact
2941 tcgccagcgc tgcactaccc atccccgaag tgttggacat cggcgagttc agcgagagcc
3001 tgacatactg catcagtaga cgcgcccaag gcgttactct ccaagacctc cccgaaacag
3061 agctgcctgc tgtgttacag cctgtcgccg aagctatgga tgctattgcc gccgccgacc
3121 tcagtcaaac cagcggcttc ggcccattcg ggccccaagg catcggccag tacacaacct
3181 ggcgggattt catttgcgcc attgctgatc cccatgtcta ccactggcag accgtgatgg
3241 acgacaccgt gtccgccagc gtagctcaag ccctggacga actgatgctg tgggccgaag
3301 actgtcccga ggtgcgccac ctcgtccatg ccgacttcgg cagcaacaac gtcctgaccg
3361 acaacggccg catcaccgcc gtaatcgact ggtccgaagc tatgttcggg gacagtcagt
3421 acgaggtggc caacatcttc ttctggcggc cctggctggc ttgcatggag cagcagactc
3481 gctacttcga gcgccggcat cccgagctgg ccggcagccc tcgtctgcga gcctacatgc
3541 tgcgcatcgg cctggatcag ctctaccaga gcctcgtgga cggcaacttc gacgatgctg
3601 cctgggctca aggccgctgc gatgccatcg tccgcagcgg ggccggcacc gtcggtcgca
3661 cacaaatcgc tcgccggagc gcagccgtat ggaccgacgg ctgcgtcgag gtgctggccg
3721 acagcggcaa ccgccggccc agtacacgac cgcgcgctaa ggaggtaggt cgagtttaaa
3781 ctctagaacc ggtcatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
3841 ttttttgtgt gttcgaacta gatgctgtcg accgatgccc ttgagagcct tcaacccagt
3901 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
3961 tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact
4021 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
4081 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
4141 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
4201 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
4261 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
4321 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
4381 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
4441 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
4501 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
4561 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
4621 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
4681 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
4741 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
4801 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
4861 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
4921 aaacttggtc tgacagcggc cgcaaatgct aaaccactgc agtggttacc agtgcttgat
4981 cagtgaggca ccgatctcag cgatctgcct atttcgttcg tccatagtgg cctgactccc
5041 cgtcgtgtag atcactacga ttcgtgaggg cttaccatca ggccccagcg cagcaatgat
5101 gccgcgagag ccgcgttcac cggcccccga tttgtcagca atgaaccagc cagcagggag
5161 ggccgagcga agaagtggtc ctgctacttt gtccgcctcc atccagtcta tgagctgctg
5221 tcgtgatgct agagtaagaa gttcgccagt gagtagtttc cgaagagttg tggccattgc
5281 tactggcatc gtggtatcac gctcgtcgtt cggtatggct tcgttcaact ctggttccca
5341 gcggtcaagc cgggtcacat gatcacccat attatgaaga aatgcagtca gctccttagg
5401 gcctccgatc gttgtcagaa gtaagttggc cgcggtgttg tcgctcatgg taatggcagc
5461 actacacaat tctcttaccg tcatgccatc cgtaagatgc ttttccgtga ccggcgagta
5521 ctcaaccaag tcgttttgtg agtagtgtat acggcgacca agctgctctt gcccggcgtc
5581 tatacgggac aacaccgcgc cacatagcag tactttgaaa gtgctcatca tcgggaatcg
5641 ttcttcgggg cggaaagact caaggatctt gccgctattg agatccagtt cgatatagcc
5701 cactcttgca cccagttgat cttcagcatc ttttactttc accagcgttt cggggtgtgc
5761 aaaaacaggc aagcaaaatg ccgcaaagaa gggaatgagt gcgacacgaa aatgttggat
5821 gctcatactc gtcctttttc aatattattg aagcatttat cagggttact agtacgtctc
5881 tcaaggataa gtaagtaata ttaaggtacg ggaggtattg gacaggccgc aataaaatat
5941 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg
6001 ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa
6061 gtgcaggtgc cagaacattt ctct
//
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