| 质粒名称: | pREP-8xARE-GFP-SV40-BFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 134910 |
| 存储实验室: | Markus Landthaler |
|
载体骨架基本信息 |
|
| 载体名称: | pMIR |
| 载体抗性: | 氨苄青霉素 |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | 8xARE- minimal promoter - GFP |
| 物种来源: | Synthetic |
| 启动子 | 8xARE - minimal promoter |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | NdeI (未破坏) |
| 5' 测序引物 | CGGATAACAATTTCACACAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pREP-8xARE-GFP-SV40-BFP was a gift from Markus Landthaler (Addgene plasmid # 134910 ; http://n2t.net/addgene:134910 ; RRID:Addgene_134910) |
| 参考文献部分: | Single-cell RNA-sequencing of herpes simplex virus 1-infected cells connects NRF2 activation to an antiviral program. Wyler E, Franke V, Menegatti J, Kocks C, Boltengagen A, Praktiknjo S, Walch-Ruckheim B, Bosse J, Rajewsky N, Grasser F, Akalin A, Landthaler M. Nat Commun. 2019 Oct 25;10(1):4878. doi: 10.1038/s41467-019-12894-z. 10.1038/s41467-019-12894-z PubMed 31653857 |
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 pREP-8xARE-GFP-SV40-BFP 6383 bp ds-DNA circular SYN 09-JAN-2020
DEFINITION NRF2 reporter plasmid with GFP under control of 8 ARE elements and
constitutively transcribed BFP.
ACCESSION .
VERSION .
KEYWORDS pREP-8xARE-GFP-SV40-BFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6383)
TITLE Single-cell RNA-sequencing of Herpes simplex virus 1-infected cells
connects NRF2 activation to an antiviral program
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6383)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 9, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6383
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 1..92
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 41..60
/label=RVprimer3
/note="pGL3 vector, forward primer"
promoter 221..252
/label=minP
/note="minimal TATA-box promoter with low basal activity"
regulatory 285..294
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 291..1010
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(336..357)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(597..616)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 944..965
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1029..1048)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1148..1170
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
promoter 1210..1526
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1377..1512
/label=SV40 ori
/note="SV40 origin of replication"
CDS 1623..2222
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(1623..1642)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2119..2139
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 2452..2533
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 2786..3143
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2809..2829)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2994..3129
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3165..3866
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
DLPSKLGHKLN"
polyA_signal 3890..4011
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3901..3920)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
CDS 4172..5032
/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 complement(4390..4409)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5203..5791
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5692..5711
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5945..5962
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6079..6100
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6115..6145
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6153..6169
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 6158..6180
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6177..6193
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6177..6193
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 6213..6231
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 6213..6230
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 6243..6262
/label=T7
/note="T7 promoter, forward primer"
promoter 6243..6261
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 6322..6370
/note="synthetic polyadenylation signal"
ORIGIN
1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
61 cagtgcaagt gcaggtgcca gaacatttct ctggcctaac tggccgtgac actgcagcag
121 tgactacgca tgcgtgacga tgcagacgtg acactgcact ggtgacaaag cacccgtgac
181 taggcaagcg tgacgtagca tcggtgacac agcaagacac tagagggtat ataatggaag
241 ctcgacttcc agcttggcaa tccggtactg ttggtaaact cgaggccacc atggtgagca
301 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
361 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
421 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
481 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
541 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
601 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
661 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
721 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
781 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
841 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
901 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
961 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa catatggtgc
1021 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
1081 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
1141 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
1201 cgaaagattg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
1261 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
1321 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
1381 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
1441 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
1501 tttttggagg cctaggcttt tgcaaaaagc tagcttgcat gcctgcaggt cggccgccac
1561 gaccggtgcc gccaccatcc cctgacccac gcccctgacc cctcacaagg agacgacctt
1621 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
1681 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
1741 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
1801 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
1861 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
1921 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
1981 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
2041 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
2101 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
2161 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
2221 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctccg
2281 accgaagcca cccggggcgg ccccgccgac cccgcacccg cccccgaggc ccaccgactc
2341 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
2401 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
2461 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
2521 tttttttcac tgcaatctcg tgatacgcct atttttatag gttaatgtca tgataataat
2581 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
2641 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
2701 tcaataatat ctcgtcatgt gggcccatct gcgcagcacc atggcctgaa ataacctctg
2761 aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
2821 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
2881 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
2941 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
3001 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
3061 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
3121 ttggaggcct aggcttttgc aaaaagcttg attcttctga cacgatgagc gagctgatta
3181 aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat cacttcaagt
3241 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtgg
3301 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc ctctacggca
3361 gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag tccttccctg
3421 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
3481 aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact
3541 tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga
3601 cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg aagctcgtgg
3661 gcgggagcca tctgatcgca aacatcaaga ccacatatag atccaagaaa cccgctaaga
3721 acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc aaggaggcca
3781 acaacgagac ctacgtcgag cagcacgagg tggcagtggc cagatactgc gacctcccta
3841 gcaaactggg gcacaagctt aattaagcgg ccgcttcgag cagacatgat aagatacatt
3901 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
3961 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
4021 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
4081 taaaacctct acaaatgtgg taaaatcgat aaggatcgat ccaggtggca ctttgggccc
4141 atctgctgtc atatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
4201 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
4261 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
4321 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
4381 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
4441 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
4501 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
4561 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
4621 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
4681 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
4741 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
4801 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
4861 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
4921 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
4981 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
5041 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
5101 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
5161 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
5221 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttttttgcc
5281 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
5341 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
5401 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
5461 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
5521 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
5581 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
5641 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
5701 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
5761 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
5821 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
5881 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
5941 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
6001 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
6061 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
6121 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
6181 aaacagctat gacatgatta cgaattgcaa cgatttaggt gacactatag aagagaagga
6241 attaatacga ctcactatag ggagagagag agaattaccc tcactaaagg gaggagaagc
6301 atgaattcgg cctaactggc caataaaata tctttatttt cattacatct gtgtgttggt
6361 tttttgtgtg aatcgatagt act
//
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