| 质粒名称: | pGL4-ERSE2-luc2P-Hygro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101790 |
| 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: | 6096bp |
| 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-II |
| 物种来源: | Synthetic |
| 插入大小： | 105bp |
| 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 | raacagtaccggattgccaag |
|
文献引用方法 |
|
| 材料和方法部分: | pGL4-ERSE2-luc2P-Hygro was a gift from Seiichi Oyadomari (Addgene plasmid # 101790 ; http://n2t.net/addgene:101790 ; RRID:Addgene_101790) |
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-ERSE2-luc2P-Hygro 6096 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Luciferase reporter plasmid containing three tandem repeats of the
ER stress response element-II (ERSE-II).
ACCESSION .
VERSION .
KEYWORDS pGL4-ERSE2-luc2P-Hygro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6096)
AUTHORS Seiichi Oyadomari
TITLE Oyadomari Lab Luciferase reporters
JOURNAL unpublished
REFERENCE 2 (bases 1 to 6096)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6096
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 163..194
/label=minP
/note="minimal TATA-box promoter with low basal activity"
regulatory 221..230
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 227..1876
/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 1880..1999
/codon_start=1
/product="PEST degradation sequence from mouse ornithine
decarboxylase"
/label=hPEST
/note="human codon-optimized"
/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
polyA_signal 2051..2172
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2062..2081)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2116..2135
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 2366..2723
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2389..2409)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2574..2709
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2636..2655
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2754..3791
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
EVGRV"
RBS 3770..3778
/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 3815..3863
/note="synthetic polyadenylation signal"
primer_bind complement(4019..4036)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4190..4778)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4270..4289)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4978..5838)
/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 5943..5991
/note="synthetic polyadenylation signal"
misc_feature 6005..6096
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 6045..6064
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 ggcctaactg gccggtacct gagctcgcta gcggacgccg attgggccac gttgggagag
61 tgcctggacg ccgattgggc cacgttggga gagtgcctgg acgccgattg ggccacgttg
121 ggagagtgcc tagatctggc ctcggcggcc aagcttagac actagagggt atataatgga
181 agctcgactt ccagcttggc aatccggtac tgttggtaaa gccaccatgg aagatgccaa
241 aaacattaag aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca
301 gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc
361 acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga
421 agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag
481 cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc
541 taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt
601 cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat
661 catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta
721 caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag
781 cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc
841 caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc
901 catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca
961 cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat
1021 gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc
1081 cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga
1141 cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga
1201 ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac
1261 aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt
1321 ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa
1381 ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc
1441 cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta
1501 ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa
1561 gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt
1621 cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt
1681 cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca
1741 ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg
1801 actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg
1861 cggcaagatc gccgtgaatt ctcacggctt ccctcccgag gtggaggagc aggccgccgg
1921 caccctgccc atgagctgcg cccaggagag cggcatggat agacaccctg ctgcttgcgc
1981 cagcgccagg atcaacgtct aaggccgcga ctctagagtc ggggcggccg gccgcttcga
2041 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
2101 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
2161 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
2221 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg
2281 tttgcgtatt gggcgctctt ccgctgatct gcgcagcacc atggcctgaa ataacctctg
2341 aaagaggaac ttggttagct accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
2401 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
2461 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
2521 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
2581 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
2641 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
2701 ttggaggcct aggcttttgc aaaaagctcg attcttctga cactagcgcc accatgaaga
2761 agcccgaact caccgctacc agcgttgaaa aatttctcat cgagaagttc gacagtgtga
2821 gcgacctgat gcagttgtcg gagggcgaag agagccgagc cttcagcttc gatgtcggcg
2881 gacgcggcta tgtactgcgg gtgaatagct gcgctgatgg cttctacaaa gaccgctacg
2941 tgtaccgcca cttcgccagc gctgcactac ccatccccga agtgttggac atcggcgagt
3001 tcagcgagag cctgacatac tgcatcagta gacgcgccca aggcgttact ctccaagacc
3061 tccccgaaac agagctgcct gctgtgttac agcctgtcgc cgaagctatg gatgctattg
3121 ccgccgccga cctcagtcaa accagcggct tcggcccatt cgggccccaa ggcatcggcc
3181 agtacacaac ctggcgggat ttcatttgcg ccattgctga tccccatgtc taccactggc
3241 agaccgtgat ggacgacacc gtgtccgcca gcgtagctca agccctggac gaactgatgc
3301 tgtgggccga agactgtccc gaggtgcgcc acctcgtcca tgccgacttc ggcagcaaca
3361 acgtcctgac cgacaacggc cgcatcaccg ccgtaatcga ctggtccgaa gctatgttcg
3421 gggacagtca gtacgaggtg gccaacatct tcttctggcg gccctggctg gcttgcatgg
3481 agcagcagac tcgctacttc gagcgccggc atcccgagct ggccggcagc cctcgtctgc
3541 gagcctacat gctgcgcatc ggcctggatc agctctacca gagcctcgtg gacggcaact
3601 tcgacgatgc tgcctgggct caaggccgct gcgatgccat cgtccgcagc ggggccggca
3661 ccgtcggtcg cacacaaatc gctcgccgga gcgcagccgt atggaccgac ggctgcgtcg
3721 aggtgctggc cgacagcggc aaccgccggc ccagtacacg accgcgcgct aaggaggtag
3781 gtcgagttta aactctagaa ccggtcatgg ccgcaataaa atatctttat tttcattaca
3841 tctgtgtgtt ggttttttgt gtgttcgaac tagatgctgt cgaccgatgc ccttgagagc
3901 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
3961 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
4021 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
4081 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
4141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
4201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
4261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
4321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
4381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
4441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
4501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
4561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
4621 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
4681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
4741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
4801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
4861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
4921 gtatatatga gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact gcagtggtta
4981 ccagtgcttg atcagtgagg caccgatctc agcgatctgc ctatttcgtt cgtccatagt
5041 ggcctgactc cccgtcgtgt agatcactac gattcgtgag ggcttaccat caggccccag
5101 cgcagcaatg atgccgcgag agccgcgttc accggccccc gatttgtcag caatgaacca
5161 gccagcaggg agggccgagc gaagaagtgg tcctgctact ttgtccgcct ccatccagtc
5221 tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt tccgaagagt
5281 tgtggccatt gctactggca tcgtggtatc acgctcgtcg ttcggtatgg cttcgttcaa
5341 ctctggttcc cagcggtcaa gccgggtcac atgatcaccc atattatgaa gaaatgcagt
5401 cagctcctta gggcctccga tcgttgtcag aagtaagttg gccgcggtgt tgtcgctcat
5461 ggtaatggca gcactacaca attctcttac cgtcatgcca tccgtaagat gcttttccgt
5521 gaccggcgag tactcaacca agtcgttttg tgagtagtgt atacggcgac caagctgctc
5581 ttgcccggcg tctatacggg acaacaccgc gccacatagc agtactttga aagtgctcat
5641 catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat tgagatccag
5701 ttcgatatag cccactcttg cacccagttg atcttcagca tcttttactt tcaccagcgt
5761 ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga gtgcgacacg
5821 aaaatgttgg atgctcatac tcgtcctttt tcaatattat tgaagcattt atcagggtta
5881 ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat tggacaggcc
5941 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
6001 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
6061 tccccagtgc aagtgcaggt gccagaacat ttctct
//
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