| 质粒名称: | pcDNA3-N-Flag-NLRP3 1-90 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 75137 |
| 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: | 5722bp |
| 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: | 276bp |
| 基因突变: | fragment encoding amino acids 1-90 |
| GenBank编号 | NM_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 1-90 was a gift from Bruce Beutler (Addgene plasmid # 75137 ; http://n2t.net/addgene:75137 ; RRID:Addgene_75137)
|
| 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
|
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 pcDNA3-N-Flag-NLRP3 1-90 5709 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses N-terminal Flag-tagged amino acids 1-90 of NLRP3 (the
pyrin domain) in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pcDNA3-N-Flag-NLRP3 1-90
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5709)
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 5709)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5709
/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(1261..1279)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1262..1279)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1299..1316)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1305..1529
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1575..2003
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1662..1681)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1872..1893
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2012..2032)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2017..2347
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2198..2333
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2260..2279
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2414..3208
/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(2468..2487)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3078..3097
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3382..3503
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3419..3438)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3473..3492
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3552..3568)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3552..3568)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3565..3587)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3576..3592
/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(3600..3630)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3645..3666
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3783..3800)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3954..4542)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4034..4053)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4713..5573)
/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 5336..5355
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5574..5678)
/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 atgacgagtg tccgttgcaa gctggctcag tatctagagg accttgaaga tgtggacctc
1021 aagaaattca aaatgcattt ggaagattac ccgcccgaga aaggctgtat cccagtcccc
1081 aggggccaga tggagaaggc agatcacttg gatctagcca cactcatgat tgacttcaat
1141 ggtgaggaga aggcctgggc catggctgtg tggatctttg ctgcgatcaa caggcgagac
1201 ctctgggaaa aagctaagaa ggaccagcca tagctcgagc atgcatctag agggccctat
1261 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
1321 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
1381 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
1441 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
1501 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
1561 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
1621 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
1681 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
1741 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
1801 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
1861 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
1921 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
1981 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
2041 tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
2101 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
2161 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
2221 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
2281 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
2341 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
2401 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
2461 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
2521 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
2581 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
2641 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
2701 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
2761 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
2821 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
2881 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
2941 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
3001 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
3061 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
3121 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
3181 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
3241 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
3301 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
3361 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
3421 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
3481 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
3541 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
3601 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
3661 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
3721 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
3781 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
3841 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
3901 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
3961 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
4021 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
4081 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
4141 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
4201 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
4261 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
4321 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
4381 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
4441 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
4501 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
4561 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
4621 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
4681 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
4741 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
4801 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
4861 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
4921 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
4981 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
5041 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
5101 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
5161 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
5221 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
5281 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
5341 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
5401 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
5461 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
5521 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
5581 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
5641 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
5701 cctgacgtc
//
