| 质粒名称: | pcDNA3-HA-mRipk3 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 78805 |
| 存储实验室: | Jaewhan Song |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3-HA |
| 完整质粒大小: | 6885 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | RIPK3 |
| 物种来源: | M. musculus (mouse) |
| 基因突变: | Ripk3 (a.k.a. 2610528K09Rik, Rip3) |
| 启动子: | CMV |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知(未破坏) |
| 3’克隆位点: | 未知 (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGHrev |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3-HA-mRipk3 was a gift from Jaewhan Song (Addgene plasmid # 78805 ; http://n2t.net/addgene:78805 ; RRID:Addgene_78805) |
| 参考文献部分: | CHIP controls necroptosis through ubiquitylation- and lysosome-dependent degradation of RIPK3. Seo J, Lee EW, Sung H, Seong D, Dondelinger Y, Shin J, Jeong M, Lee HK, Kim JH, Han SY, Lee C, Seong JK, Vandenabeele P, Song J. Nat Cell Biol. 2016 Mar;18(3):291-302. doi: 10.1038/ncb3314. Epub 2016 Feb 22. 10.1038/ncb3314 PubMed 26900751 |
买家导航
pcDNA3-HA-mRipk3载体质粒基本信息
pcDNA3-HA-mRipk3质粒图谱载体图谱和pcDNA3-HA-mRipk3载体序列质粒序列多克隆位点信息
pcDNA3-HA-mRipk3质粒载体简介
pcDNA3-HA-mRipk3质粒序列载体序列
LOCUS pcDNA3-HA-mRipk3 6885 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION To overexpress mRipk3 in Mammalian Cells.
ACCESSION .
VERSION .
KEYWORDS pcDNA3-HA-mRipk3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6885)
AUTHORS Seo J, Lee EW, Sung H, Seong D, Dondelinger Y, Shin J, Jeong M, Lee
HK, Kim JH, Han SY, Lee C, Seong JK, Vandenabeele P, Song J
TITLE CHIP controls necroptosis through ubiquitylation- and
lysosome-dependent degradation of RIPK3.
JOURNAL Nat Cell Biol. 2016 Mar;18(3):291-302. doi: 10.1038/ncb3314. Epub
2016 Feb 22.
PUBMED 26900751
REFERENCE 2 (bases 1 to 6885)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6885
/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 910..936
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2298..2309
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
promoter complement(2437..2455)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2438..2455)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2475..2492)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2481..2705
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2751..3179
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2838..2857)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3048..3069
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3188..3208)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3193..3523
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3374..3509
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3436..3455
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3590..4384
/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(3644..3663)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4254..4273
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4558..4679
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4595..4614)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4649..4668
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4728..4744)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4728..4744)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4741..4763)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4752..4768
/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(4776..4806)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4821..4842
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4959..4976)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5130..5718)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5210..5229)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5889..6749)
/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 6512..6531
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6750..6854)
/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 atggcctcct acccttatga tgtgccagat tatgcctctc ccgaattcat gtcttctgtc
961 aagttatggc ctactggtgc gtcagcggtt cctctggtga gccgtgaaga actgaagaag
1021 ctggagtttg tgggcaaagg agggttcgga gtcgtgttcc gggcacacca cagaacatgg
1081 aaccatgatg tagcagtcaa gatcgtgaac tcgaagaaga tatcctggga ggtgaaggct
1141 atggttaatc ttcgtaatga gaacgttctg ctcctgctgg gggtcactga ggacctccag
1201 tgggacttcg tgtccgggca ggctctggtg acaagattca tggagaatgg ctccctcgca
1261 gggctgctgc aacccgagtg ccctcggccc tggccactcc tctgtcgcct gctgcaggaa
1321 gtggtgctgg ggatgtgcta cctacacagc ttgaaccctc cgctcctgca ccgggacctc
1381 aagccctcta acattctgct ggatccagag ctccacgcca agctagcaga ttttggcctg
1441 tccacgtttc agggagggtc ccagtcaggg tcaggatcag gatcaggatc cagggactct
1501 gggggcaccc tagcgtactt ggacccagag ctgttattta atgtcaacct gaaggcttct
1561 aaagcgagtg atgtctacag ctttgggatc ctcgtgtggg cagtgctggc tggcagagaa
1621 gctgagttgg tagacaagac ttcactaatc cgggaaacag tgtgtgacag gcagagtcgt
1681 cctccactga cagagctgcc tccaggtagc cctgagactc ctggcttgga aaaactgaag
1741 gagttaatga ttcattgctg gggttcccag tccgaaaaca ggccatcctt ccaggactgc
1801 gaaccaaaaa ccaatgaagt ttacaatctg gtaaaggaca aggtagatgc tgctgtctcc
1861 gaggtaaagc attatctgtc tcagcacaga agcagcggca gaaacttgtc tgccagagag
1921 ccaagccaaa gaggcacaga aatggattgc ccgagggaaa ccatggtttc taaaatgctg
1981 gaccgcctgc atttggagga accctccgga ccagttcctg gaaaatgtcc tgagaggcaa
2041 gcacaggaca catcagttgg gcctgccaca ccagcaagga catcttctga ccccgtggct
2101 ggcactcctc agattccaca tactttaccc ttcagaggca caacacctgg gccagtcttt
2161 actgagactc ccggtcctca cccccaaagg aatcagggag atggaagaca cggcactcct
2221 tggtatccct ggaccccacc gaatccaatg acagggccac cggctctcgt cttcaacaac
2281 tgttctgaag tgcagattgg gaactacaac tccttggtag caccaccaag aactactgcc
2341 tcaagttcgg ccaagtatga ccaagcacag ttcggcaggg gtaggggctg gcagcccttc
2401 cacaagtagc tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
2461 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
2521 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
2581 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
2641 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
2701 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
2761 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
2821 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
2881 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
2941 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
3001 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
3061 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
3121 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
3181 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag
3241 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
3301 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
3361 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
3421 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
3481 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
3541 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
3601 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
3661 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
3721 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
3781 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
3841 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
3901 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
3961 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
4021 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
4081 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
4141 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
4201 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
4261 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
4321 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
4381 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
4441 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
4501 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
4561 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
4621 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
4681 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
4741 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
4801 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
4861 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
4921 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
4981 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
5041 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
5101 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
5161 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
5221 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
5281 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
5341 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
5401 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
5461 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
5521 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
5581 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
5641 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
5701 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
5761 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
5821 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
5881 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
5941 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
6001 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
6061 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
6121 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
6181 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
6241 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
6301 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
6361 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
6421 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
6481 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
6541 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
6601 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
6661 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
6721 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
6781 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
6841 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
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