| 質粒名稱: | pcDNA3.1-HA-NONO |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 127655 |
| 存儲實驗室: | Nicolas Manel |
|
載體骨架基本信息 |
|
| 載體名稱: | pcDNA3.1 |
| 載體出品公司: | Clontech |
| 空載體大小: | 4038 bp |
| 完整質粒大小: | 6840bp |
| 載體類型: | 哺乳細胞表達 |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | NONO |
| 物種來源: | H. sapiens (human) |
| 標簽/融合蛋白: | HA (N terminal on insert) |
| 克隆方法: | 限制性內切酶 |
| 5’克隆位點: | AgeI(未破壞) |
| 3’克隆位點: | NotI (未破壞) |
| 5’測序引物 | CMV-F |
| 3’測序引物 | SV40pA-R |
|
文獻引用方法 |
|
| 材料和方法部分: | pcDNA3.1-HA-NONO was a gift from Nicolas Manel (Addgene plasmid # 127655 ; //n2t.net/addgene:127655 ; RRID:Addgene_127655) |
| 參考文獻部分: | NONO Detects the Nuclear HIV Capsid to Promote cGAS-Mediated Innate Immune Activation. Lahaye X, Gentili M, Silvin A, Conrad C, Picard L, Jouve M, Zueva E, Maurin M, Nadalin F, Knott GJ, Zhao B, Du F, Rio M, Amiel J, Fox AH, Li P, Etienne L, Bond CS, Colleaux L, Manel N. Cell. 2018 Oct 4;175(2):488-501.e22. doi: 10.1016/j.cell.2018.08.062. Epub 2018 Sep 27. 10.1016/j.cell.2018.08.062 PubMed 30270045 |
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pcDNA3.1-HA-NONO載體質粒基本信息
pcDNA3.1-HA-NONO質粒圖譜載體圖譜和pcDNA3.1-HA-NONO載體序列質粒序列多克隆位點信息
pcDNA3.1-HA-NONO質粒載體簡介
pcDNA3.1-HA-NONO質粒序列載體序列
LOCUS pcDNA3.1-HA-NONO 6840 bp DNA circular SYN 14-APR-2025
DEFINITION Over-express HA-tagged human NONO.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6840)
AUTHORS Lahaye X, Gentili M, Silvin A, Conrad C, Picard L, Jouve M, Zueva E,
Maurin M, Nadalin F, Knott GJ, Zhao B, Du F, Rio M, Amiel J, Fox AH,
Li P, Etienne L, Bond CS, Colleaux L, Manel N
TITLE NONO Detects the Nuclear HIV Capsid to Promote cGAS-Mediated Innate
Immune Activation.
JOURNAL Cell. 2018 Oct 4;175(2):488-501.e22. doi:
10.1016/j.cell.2018.08.062. Epub 2018 Sep 27.
PUBMED 30270045
REFERENCE 2 (bases 1 to 6840)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2018.08.062"; journalName: "Cell"; date: "2018-10-4-
4"; volume: "175"; issue: "2"; pages: "488-501.e22"
COMMENT Sequence Label: pcDNA3.1-HA-NONO
FEATURES Location/Qualifiers
source 1..6840
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 908..934
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(2393..2411)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2394..2411)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2431..2448)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2437..2661
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2707..3135
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2794..2813)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3004..3025
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3144..3164)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3149..3478
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3329..3464
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3391..3410
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3545..4339
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3599..3618)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4209..4228
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4513..4634
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4550..4569)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4604..4623
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4683..4699)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4683..4699)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4696..4718)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4707..4723
/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)."
promoter complement(join(4731..4737,4738..4755,4756..4761))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4776..4797
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4914..4931)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5085..5673)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5165..5184)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(5844..6635,6636..6704))
/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 6467..6486
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6705..6809)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttagccac
901 caatatgtac ccatacgatg ttccagatta cgctggagca agggatccaa tgcagagtaa
961 taaaactttt aacttggaga agcaaaacca tactccaaga aagcatcatc aacatcacca
1021 ccagcagcag caccaccagc agcaacagca gcagccgcca ccaccgccaa tacctgcaaa
1081 tgggcaacag gccagcagcc aaaatgaagg cttgactatt gacctgaaga attttagaaa
1141 accaggagag aagaccttca cccaacgaag ccgtcttttt gtgggaaatc ttcctcccga
1201 catcactgag gaagaaatga ggaaactatt tgagaaatat ggaaaggcag gcgaagtctt
1261 cattcataag gataaaggat ttggctttat ccgcttggaa acccgaaccc tagcggagat
1321 tgccaaagtg gagctggaca atatgccact ccgtggaaag cagctgcgtg tgcgctttgc
1381 ctgccatagt gcatccctta cagttcgaaa ccttcctcag tatgtgtcca acgaactgct
1441 ggaagaagcc ttttctgtgt ttggccaggt agagagggct gtagtcattg tggatgatcg
1501 aggaaggccc tcaggaaaag gcattgttga gttctcaggg aagccagctg ctcggaaagc
1561 tctggacaga tgcagtgaag gctccttcct gctaaccaca tttcctcgtc ctgtgactgt
1621 ggagcccatg gaccagttag atgatgaaga gggacttcca gagaagctgg ttataaaaaa
1681 ccagcaattt cacaaggaac gagagcagcc acccagattt gcacagcctg gctcctttga
1741 gtatgaatat gccatgcgct ggaaggcact cattgagatg gagaagcagc agcaggacca
1801 agtggaccgc aacatcaagg aggctcgtga gaagctggag atggagatgg aagctgcacg
1861 ccatgagcac caggtcatgc taatgagaca ggatttgatg aggcgccaag aagaacttcg
1921 gaggatggaa gagctgcaca accaagaggt gcaaaaacga aagcaactgg agctcaggca
1981 ggaggaagag cgcaggcgcc gtgaagaaga gatgcggcgg cagcaagaag aaatgatgcg
2041 gcgacagcag gaaggattca agggaacctt ccctgatgcg agagagcagg agattcggat
2101 gggtcagatg gctatgggag gtgctatggg cataaacaac agaggtgcca tgccccctgc
2161 tcctgtgcca gctggtaccc cagctcctcc aggacctgcc actatgatgc cggatggaac
2221 tttgggattg accccaccaa caactgaacg ctttggtcag gctgctacaa tggaaggaat
2281 tggggcaatt ggtggaactc ctcctgcatt caaccgtgca gctcctggag ctgaatttgt
2341 cccaaacaaa cgtcgccgat actaactcga gcatgcatct agagggccct attctatagt
2401 gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
2461 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
2521 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
2581 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
2641 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
2701 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
2761 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
2821 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
2881 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
2941 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
3001 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
3061 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
3121 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
3181 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
3241 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
3301 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
3361 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
3421 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
3481 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
3541 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
3601 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
3661 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
3721 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
3781 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
3841 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
3901 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
3961 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
4021 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
4081 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
4141 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
4201 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
4261 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
4321 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
4381 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
4441 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
4501 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
4561 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
4621 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
4681 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
4741 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
4801 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
4861 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
4921 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
4981 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
5041 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
5101 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
5161 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
5221 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
5281 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
5341 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
5401 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
5461 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
5521 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
5581 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
5641 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
5701 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
5761 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
5821 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
5881 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
5941 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
6001 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
6061 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
6121 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
6181 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
6241 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
6301 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
6361 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
6421 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
6481 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
6541 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
6601 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
6661 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
6721 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
6781 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//