| 质粒名称: | pcDNA3-N-Flag-NLRP3 711-1034 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 75141 |
| 存储实验室: | Bruce Beutler |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | Clontech |
| 空载体大小: | 5446bp |
| 完整质粒大小: | 6411bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | NLR family, pyrin domain containing 3 / Nlrp3 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 965bp |
| 基因突变: | fragment encoding amino acids 711-1034 |
| GenBank编号 | JNM_145827 |
| 启动子: | CMV |
| 标签/融合蛋白: | Flag (N terminal on insert) |
| 克隆方法: | 未知 |
| 5’测序引物 | CMV-F |
|
文献引用方法 |
|
| 材料和方法部分: |
pcDNA3-N-Flag-NLRP3 711-1034 was a gift from Bruce Beutler (Addgene plasmid # 75141 ; http://n2t.net/addgene:75141 ; RRID:Addgene_75141)
|
| 参考文献部分: | NLRP3 activation and mitosis are mutually exclusive events coordinated by NEK7, a new inflammasome component. Shi H, Wang Y, Li X, Zhan X, Tang M, Fina M, Su L, Pratt D, Bu CH, Hildebrand S, Lyon S, Scott L, Quan J, Sun Q, Russell J, Arnett S, Jurek P, Chen D, Kravchenko VV, Mathison JC, Moresco EM, Monson NL, Ulevitch RJ, Beutler B. Nat Immunol. 2016 Mar;17(3):250-8. doi: 10.1038/ni.3333. Epub 2015 Dec 7. 10.1038/ni.3333 PubMed 26642356 |
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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 pcDNA3-N-Flag-NLRP3 711-1034 6408 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses N-terminal Flag-tagged amino acids 711-1034 of NLRP3 (the
LRR domain) in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pcDNA3-N-Flag-NLRP3 711-1034
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6408)
AUTHORS Shi H, Wang Y, Li X, Zhan X, Tang M, Fina M, Su L, Pratt D, Bu CH,
Hildebrand S, Lyon S, Scott L, Quan J, Sun Q, Russell J, Arnett S,
Jurek P, Chen D, Kravchenko VV, Mathison JC, Moresco EM, Monson NL,
Ulevitch RJ, Beutler B
TITLE NLRP3 activation and mitosis are mutually exclusive events
coordinated by NEK7, a new inflammasome component.
JOURNAL Nat Immunol. 2016 Mar;17(3):250-8. doi: 10.1038/ni.3333. Epub 2015
Dec 7.
PUBMED 26642356
REFERENCE 2 (bases 1 to 6408)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6408
/organism="synthetic DNA construct"
/mol_type="other DNA"
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 922..945
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(1960..1978)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1961..1978)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1998..2015)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2004..2228
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2274..2702
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2361..2380)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2571..2592
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2711..2731)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2716..3046
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2897..3032
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2959..2978
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3113..3907
/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(3167..3186)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3777..3796
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4081..4202
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4118..4137)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4172..4191
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4251..4267)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4251..4267)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4264..4286)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4275..4291
/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(4299..4329)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4344..4365
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4482..4499)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4653..5241)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4733..4752)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5412..6272)
/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 6035..6054
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6273..6377)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
901 gagctcggat cgcccaccat ggactacaaa gacgatgacg ataaaatcga tatcgaattc
961 gttgcctgtt cttccagact ggtgaactgc tgcctcactt ctagcttctg ccgtggtctc
1021 ttctcaagtc taagcaccaa ccggagcctc actgaactgg acctcagtga caatactctg
1081 ggagacccgg gcatgagggt gctgtgtgag gcactccagc acccaggctg taacattcag
1141 agactgtggt tggggcgctg cggactgtcc catcaatgct gcttcgacat ctcctctgtc
1201 ctgagcagca gccagaagct ggtggagctg gacctcagtg acaatgccct gggggacttt
1261 ggaatcagat tgctgtgtgt gggactgaag cacctgctct gcaacctcca gaaactgtgg
1321 ttggtgagct gctgtctcac atccgcgtgt tgtcaggatc tcgcattggt tctgagctcc
1381 aaccattctc tgaccagact gtacattgga gaaaatgcct tgggagactc aggagtccaa
1441 gttttgtgtg aaaagatgaa ggacccacag tgtaacttgc agaagctggg gttggtgaat
1501 tccggcctta cttcaatctg ttgttcagct ctgacctctg tgctcaaaac caaccagaac
1561 ttcacacacc tctatctacg aagcaatgcc cttggagaca caggactcag gctcctctgt
1621 gaggggcttc tgcacccgga ctgtaaacta cagatgctgg aattagacaa ctgcagcctc
1681 acctcacaca gctgctggaa tctctccaca attctgaccc acaaccacag ccttcggaag
1741 ctgaacctgg gcaacaatga tcttggcgat ctgtgcgtgg tgaccctctg tgaggtgctg
1801 aaacagcagg gctgcctcct gcagagccta cagttgggtg aaatgtactt aaatcgtgaa
1861 acaaaacgtg ccttagaagc gctccaggaa gaaaagcctg agctgactat agtcttcgag
1921 atttcctggt agctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
1981 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
2041 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
2101 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
2161 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
2221 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
2281 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
2341 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
2401 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
2461 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
2521 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
2581 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
2641 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
2701 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg
2761 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
2821 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
2881 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
2941 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
3001 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
3061 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
3121 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
3181 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
3241 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
3301 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
3361 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
3421 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
3481 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
3541 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
3601 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
3661 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
3721 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
3781 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
3841 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
3901 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
3961 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
4021 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
4081 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
4141 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
4201 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
4261 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
4321 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
4381 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
4441 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
4501 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
4561 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
4621 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
4681 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
4741 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
4801 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
4861 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
4921 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
4981 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
5041 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
5101 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
5161 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
5221 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
5281 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
5341 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
5401 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
5461 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
5521 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
5581 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
5641 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
5701 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
5761 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
5821 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
5881 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
5941 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
6001 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
6061 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
6121 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
6181 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
6241 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
6301 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
6361 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
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