| 质粒名称: | pαH-S-HRV3C |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 164567 |
| 存储实验室: | Priyamvada Acharya |
|
载体骨架基本信息 |
|
| 载体名称: | pαH |
| 载体出品公司: | -- |
| 空载体大小: | 4746 bp |
| 完整质粒大小: | 8358 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素 |
| 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: | Spike - Furin site changed for HRV3C site,S-HRV3C |
| 物种来源: | Severe acute respiratory syndrome coronavirus 2 |
| 插入大小: | 3636 bp |
| 基因突变: | 682-685（furin现场）替换为HRV3C现场（LEVLFQGP） |
| GenBank编号 | MN908947 |
| 启动子: | -- |
| 标签/融合蛋白: |
8X His tag（主干上的C终端）
2X Strep-Tag II（主干上的C终端）
|
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pαH-S-HRV3C was a gift from Priyamvada Acharya (Addgene plasmid # 164567 ; http://n2t.net/addgene:164567 ; RRID:Addgene_164567) |
| 参考文献部分: | D614G Mutation Alters SARS-CoV-2 Spike Conformation and Enhances Protease Cleavage at the S1/S2 Junction. Gobeil SM, Janowska K, McDowell S, Mansouri K, Parks R, Manne K, Stalls V, Kopp MF, Henderson R, Edwards RJ, Haynes BF, Acharya P. Cell Rep. 2021 Jan 12;34(2):108630. doi: 10.1016/j.celrep.2020.108630. Epub 2020 Dec 26. 10.1016/j.celrep.2020.108630 PubMed 33417835 |
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 p-alpha-H-S-HRV3C 8363 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION SARS-CoV-2 Spike protein with furin site mutated to HRV3C site
(S-HRV3C variant) .
ACCESSION .
VERSION .
KEYWORDS p-alpha-H-S-HRV3C
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8363)
AUTHORS Gobeil SM, Janowska K, McDowell S, Mansouri K, Parks R, Manne K,
Stalls V, Kopp MF, Henderson R, Edwards RJ, Haynes BF, Acharya P
TITLE D614G Mutation Alters SARS-CoV-2 Spike Conformation and Enhances
Protease Cleavage at the S1/S2 Junction.
JOURNAL Cell Rep. 2021 Jan 12;34(2):108630. doi:
10.1016/j.celrep.2020.108630. Epub 2020 Dec 26.
PUBMED 33417835
REFERENCE 2 (bases 1 to 8363)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8363
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 1601..1623
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
CDS 3808..3831
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 5500..5523
/codon_start=1
/product="8xHis affinity tag"
/label=8xHis
/translation="HHHHHHHH"
CDS 5530..5553
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 5593..5616
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
primer_bind complement(5718..5737)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 5783..5838
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(5837..5856)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(6199..6215)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6199..6215)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6212..6234)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6223..6239
/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(6247..6277)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6292..6313
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6442..6459)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6613..7201)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6693..6712)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7372..8232)
/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 7995..8014
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8233..8337)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
481 cgatgggggc gggggggggg ggggggsgsg ccvggsgggg sggggsgggg sgrggggsgg
541 ggsggggsgr ggsggrgrgg kgsggsggcr gccaatcaga gcggcgcgct ccgaaagttt
601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
1201 ggcgcggggc tcgccgtgcc gggcgggggg kggcggcrgg kgggggkgcc gggcggggcg
1261 gggccgcckc gggcsggggr gggskcgggg grggggcgcg gcggcccccg grgcgccggc
1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
1621 accggcggct ctagcgcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
1681 ctgggcaacg tgctggttgt tgtgctgtct catcattttg gcaaagaatt gcggccgtct
1741 caggccgagt tcggtaccgc caccatgttc gtgttcctgg tgctcctgcc tctggtgagc
1801 agccagtgcg tgaacctgac cacccgaacc cagctcccac cagcctacac caacagcttt
1861 acacggggcg tgtactaccc tgacaaggtg ttcagatcta gcgtcctgca cagcactcag
1921 gacctcttcc tgccgttctt cagcaacgtg acatggttcc acgccatcca cgtgagcggc
1981 acaaacggaa ccaagcggtt tgataacccc gtcctgccat tcaatgatgg agtttacttc
2041 gccagtaccg agaagagtaa catcatccgg ggctggatct tcggcaccac cctggatagc
2101 aaaacacaga gcctcctgat cgtgaacaat gccacgaacg tcgtgatcaa ggtgtgcgag
2161 ttccagtttt gcaatgatcc tttcctgggt gtgtactacc acaagaacaa caagagctgg
2221 atggaaagcg agttcagagt ctacagcagc gccaacaact gcacattcga gtacgtctct
2281 cagccttttc tgatggacct tgaggggaaa caaggcaact tcaagaacct gagagaattc
2341 gtgttcaaga acatcgacgg ctacttcaaa atctactcca agcacacacc catcaacctg
2401 gtccgggacc tccctcaggg cttcagcgcc ctggaacccc tggtcgacct gcccataggc
2461 atcaacataa cgcggttcca aaccctgctg gccctgcata gatcctacct gactcctggc
2521 gacagcagca gcggatggac cgccggagct gcagcctact atgtgggcta cctgcaacct
2581 agaaccttcc tgctgaagta caacgagaac ggcacaatca cagacgccgt cgactgcgcc
2641 ctggaccctc tctctgagac aaagtgcacc ctgaagtcct tcaccgtgga aaagggcatc
2701 taccagacca gcaacttccg ggtgcagcct acagagagca tcgtgcgatt tccaaacatt
2761 accaacctct gccccttcgg cgaggtgttt aacgccacaa gatttgcctc cgtttacgcc
2821 tggaatagaa agagaatcag caattgtgtg gccgactact ccgtgctgta taacagcgcc
2881 tctttcagca ccttcaagtg ctacggcgtt tccccaacaa agctgaatga cctgtgcttc
2941 accaacgtgt acgccgactc cttcgtaatt agaggcgatg aggtgcggca gatcgcacca
3001 ggccagaccg gtaagatcgc tgactacaac tataagctgc ctgatgattt tacaggctgc
3061 gtgatcgcct ggaactctaa caacctggat agcaaggtgg gcggcaacta caactacctg
3121 taccggctgt ttcgcaagtc taacctgaaa cctttcgaga gagacatctc cacagagatc
3181 taccaggccg gttctacacc ttgtaacggg gtggaaggct tcaactgtta cttccctctg
3241 caaagctacg gcttccagcc taccaatgga gtcggctacc agccataccg ggtggtcgtg
3301 ctgtccttcg agttactcca cgcccccgcc accgtctgcg gtcctaagaa gtccaccaat
3361 ctggttaaga acaaatgcgt gaacttcaac ttcaacggcc tgaccgggac cggcgtgctg
3421 accgaaagca acaaaaagtt cctccccttc cagcagttcg gccgtgatat cgctgacacc
3481 acagatgccg tcagagatcc acagaccctg gaaatcctgg atattacacc ctgctccttc
3541 ggaggagttt ctgtgatcac ccccgggacc aataccagca accaggtggc tgtgctgtac
3601 caagatgtta actgcaccga ggttcctgtg gccatccacg ccgatcagct gacacctact
3661 tggagagtgt actccactgg ctccaatgtg ttccagacca gggccggatg tctgatcggc
3721 gccgagcacg tgaataacag ttacgagtgc gacatcccta tcggcgccgg catctgtgcc
3781 agctaccaga cccagacaaa cagccctctg gaggtgctgt tccagggccc atctgtagct
3841 agccagagca tcatcgccta caccatgagc ctgggcgcag agaacagcgt ggcctattcc
3901 aacaactcta tcgccattcc caccaacttt acaattagcg tcacaacaga gatcctgccc
3961 gtgagcatga ccaagaccag cgtggactgt acaatgtaca tctgtggcga cagcactgaa
4021 tgcagcaacc tgctgctgca atacggctcc ttttgcaccc aactgaaccg ggcgctgacc
4081 ggaatcgccg tggaacagga caaaaatacc caggaggtgt tcgcccaagt gaagcagatc
4141 tacaagaccc cacctatcaa ggacttcggc ggctttaact ttagccagat tctccctgat
4201 ccttctaagc ctagcaagcg gagctttatc gaggatctgc tgttcaacaa ggtcaccctg
4261 gccgatgccg gctttatcaa acagtatggc gattgcctgg gcgacatagc cgccagagat
4321 ctgatctgcg cccagaaatt caacggcctg acagttctcc cacctctgct gaccgacgag
4381 atgatcgctc agtacacctc tgccctgctg gctggcacca tcacatctgg gtggacattt
4441 ggcgccggcg ccgccctgca gatccccttt gccatgcaga tggcctatag attcaacgga
4501 atcggcgtga cccagaacgt gctgtatgaa aaccagaagc tgatcgctaa ccagttcaat
4561 tctgccatcg gcaagatcca ggactccctc tcctctaccg ccagcgccct gggcaaactg
4621 caggacgtgg tgaatcagaa cgcccaagcc ctgaacaccc tggtgaagca gctcagcagc
4681 aattttggcg ccatcagctc tgtgctgaac gatatcctgt ctagactgga caaggtggaa
4741 gccgaagtcc agatcgatag actgatcaca ggcagactgc agtccctgca aacctacgtg
4801 acccaacagc tgatcagggc cgctgaaata agagccagcg ccaatctcgc cgctaccaag
4861 atgtccgagt gtgtgctggg acagtctaaa cgcgttgact tctgcggcaa aggctatcac
4921 ctgatgagct tcccccagag cgcgccgcac ggcgtggtgt tcctgcatgt gacatacgtg
4981 cctgcccaag agaagaattt cacaaccgcc cctgccatct gccacgacgg caaggcccac
5041 ttcccaagag agggcgtttt cgtttccaat ggcacacact ggttcgtgac acaaagaaac
5101 ttctacgaac cccagattat caccaccgac aacaccttcg tgagtggcaa ttgtgacgtg
5161 gtcatcggaa tcgtgaacaa cacagtgtac gaccctctgc aacctgagct ggactctttt
5221 aaggaagagc tggacaagta ctttaaaaac cacaccagcc ccgatgtgga cctgggcgac
5281 atcagtggca ttaacgccag cgtggtgaac atccaaaagg aaatcgacag actgaacgag
5341 gtggccaaga acctgaacga gtccctgatc gacctgcagg agctcggcaa atacgagcag
5401 ggatccggat acatccccga ggcccccaga gatggccagg cctacgtgcg gaaggacggc
5461 gagtgggtac tgctgagcac attcctgggc agatccggcc atcaccacca tcaccaccat
5521 catagcgcct ggtcccaccc ccagttcgag aagggcggcg gtagtggagg gggcggatct
5581 ggcggctcag cttggagcca cccccagttc gaaaagtgat aatgactcga gcgataattc
5641 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct
5701 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc
5761 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
5821 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag
5881 aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa
5941 ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc
6001 catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt
6061 tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc
6121 tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc
6181 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
6241 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
6301 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
6361 ccagcggatc gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
6421 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
6481 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
6541 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
6601 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
6661 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
6721 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
6781 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
6841 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
6901 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
6961 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
7021 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
7081 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
7141 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
7201 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
7261 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
7321 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
7381 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
7441 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
7501 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
7561 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
7621 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
7681 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
7741 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
7801 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
7861 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
7921 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
7981 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
8041 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
8101 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
8161 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
8221 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
8281 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
8341 atttccccga aaagtgccac ctg
//
