| 质粒名称: | CoV2-Spike-EF1a-D614G-N501Y |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 177939 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 完整质粒大小: | 8055 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: | SARS-CoV-2 Spike protein (D614G, N501Y) |
| 物种来源: | Other |
| 基因突变: | D614G, N501Y |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | CoV2-Spike-EF1a-D614G-N501Y was a gift from Jennifer Doudna (Addgene plasmid # 177939 ; http://n2t.net/addgene:177939 ; RRID:Addgene_177939) |
| 参考文献部分: | Rapid assessment of SARS-CoV-2 evolved variants using virus-like particles. Syed AM, Taha TY, Tabata T, Chen IP, Ciling A, Khalid MM, Sreekumar B, Chen PY, Hayashi JM, Soczek KM, Ott M, Doudna JA. Science. 2021 Nov 4:eabl6184. doi: 10.1126/science.abl6184. 10.1126/science.abl6184 PubMed 34735219 |
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 CoV2-Spike-EF1a-D614G-N501Y 8055 bp ds-DNA circular SYN 16-NOV-2021
DEFINITION Transient mammalian expression of codon optimized SARS-CoV-2 Spike
proteins (D614G,N501Y). This version gives lower expression of Spike
and is more convenient for generating VLPs..
ACCESSION .
VERSION .
KEYWORDS CoV2-Spike-EF1a-D614G-N501Y
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8055)
AUTHORS Syed AM, Taha TY, Tabata T, Chen IP, Ciling A, Khalid MM, Sreekumar
B, Chen PY, Hayashi JM, Soczek KM, Ott M, Doudna JA
TITLE Rapid assessment of SARS-CoV-2 evolved variants using virus-like
particles.
JOURNAL Science. 2021 Nov 4:eabl6184. doi: 10.1126/science.abl6184.
PUBMED 34735219
REFERENCE 2 (bases 1 to 8055)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 16, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8055
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 164..1345
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 394..1336
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1293..1313
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 5207..5795
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5260..5280)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5678..5689)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5864..5881)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5870..6094
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(6230..6818)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6310..6329)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6989..7849)
/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 7612..7631
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7850..7954)
/gene="bla"
/label=AmpR promoter
primer_bind complement(8029..8048)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
61 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
121 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtgctccgg tgcccgtcag
181 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
241 accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
301 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
361 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg
421 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac gcccctggct
481 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
541 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcttg ggcgctgggg
601 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
661 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
721 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
781 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
841 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
901 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
961 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
1021 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
1081 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
1141 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
1201 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
1261 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
1321 ttttcttcca tttcaggtgt cgtgagctag catgtttgtc ttcctggtcc tgctgcctct
1381 ggtctcgtct cagtgcgtga acctgactac tagaacccag ctgcctcctg cctatactaa
1441 ctccttcacc cgcggcgtgt actacccaga caaggtgttc cgcagctccg tgctgcactc
1501 cacccaggat ctgttcctgc ccttcttcag caacgtgacc tggttccacg ccatccacgt
1561 gagcggcacc aatggcacca agcggttcga caatcccgtg ctgccattca acgatggcgt
1621 gtacttcgcc tccaccgaga agagcaacat catccgcggc tggatcttcg gcaccaccct
1681 ggactccaag acccagagcc tgctgatcgt gaacaatgcc accaacgtgg tcatcaaggt
1741 gtgcgagttc cagttctgca atgatccatt cctgggcgtg tactaccaca agaacaataa
1801 gtcctggatg gagagcgagt tccgcgtgta cagctccgcc aacaattgca ccttcgagta
1861 cgtgtcccag cccttcctga tggacctgga gggcaagcag ggcaatttca agaacctgcg
1921 cgagttcgtg ttcaagaata tcgatggcta cttcaagatc tactccaagc acacccccat
1981 caacctggtg cgcgacctgc cacagggctt cagcgccctg gagccactgg tggatctgcc
2041 aatcggcatc aacatcacca ggttccagac cctgctggcc ctgcaccgca gctacctgac
2101 cccaggcgac agctccagcg gatggaccgc tggagctgct gcctactacg tgggctacct
2161 gcagccccgc accttcctgc tgaagtacaa cgagaatggc accatcaccg acgccgtgga
2221 ttgcgccctg gatccactgt ccgagacaaa gtgcaccctg aagagcttca ccgtggagaa
2281 gggcatctac cagacctcca atttccgcgt gcagccaacc gagagcatcg tgcgcttccc
2341 caatatcacc aacctgtgcc cattcggcga ggtgttcaac gctaccaggt tcgccagcgt
2401 gtacgcttgg aatcgcaagc gcatctccaa ctgcgtggcc gactacagcg tgctgtacaa
2461 ctccgccagc ttctccacct tcaagtgcta cggcgtgtcc cccaccaagc tgaatgatct
2521 gtgcttcacc aacgtgtacg ccgatagctt cgtgatcagg ggcgacgagg tgcgccagat
2581 cgctccagga cagaccggca agatcgctga ctacaattac aagctgcccg acgatttcac
2641 cggctgcgtg atcgcctgga actccaacaa tctggatagc aaagtgggcg gcaactacaa
2701 ttacctgtac cgcctgttcc gcaagtccaa tctgaagcca ttcgagcgcg acatctccac
2761 cgagatctac caggctggaa gcaccccatg caatggagtg gagggcttca actgctactt
2821 ccccctgcag agctacggct tccagccaac ctacggagtg ggataccagc catacagggt
2881 ggtggtgctg tccttcgagc tgctgcacgc tccagctacc gtgtgcggac caaagaagag
2941 caccaatctg gtgaagaaca agtgcgtgaa cttcaatttc aacggcctga ccggaaccgg
3001 cgtgctgacc gagtccaaca agaagttcct gccattccag cagttcggaa gggacatcgc
3061 tgataccacc gacgccgtgc gcgacccaca gaccctggag atcctggata tcaccccatg
3121 ctccttcggc ggcgtgagcg tgatcacccc aggaaccaat accagcaacc aggtggccgt
3181 gctgtaccag ggcgtgaatt gcaccgaggt gccagtggct atccacgctg atcagctgac
3241 cccaacctgg cgcgtgtaca gcaccggatc caacgtgttc cagacccgcg ccggatgcct
3301 gatcggagct gagcacgtga acaattccta cgagtgcgac atcccaatcg gagctggaat
3361 ctgcgccagc taccagaccc agaccaactc cccaaggagg gctcgcagcg tggccagcca
3421 gtccatcatc gcctacacca tgtccctggg cgccgagaat agcgtggcct acagcaacaa
3481 ttccatcgcc atcccaacca acttcaccat ctccgtgacc accgagatcc tgcccgtgtc
3541 catgaccaag accagcgtgg actgcaccat gtacatctgc ggcgattcca ccgagtgcag
3601 caacctgctg ctgcagtacg gcagcttctg cacccagctg aatcgcgccc tgaccggaat
3661 cgctgtggag caggataaga acacccagga ggtgttcgcc caggtgaagc agatctacaa
3721 gaccccccca atcaaggact tcggcggctt caatttcagc cagatcctgc ccgatccaag
3781 caagccctcc aagcgcagct tcatcgagga cctgctgttc aacaaggtga ccctggccga
3841 tgccggcttc atcaagcagt acggcgattg cctgggcgac atcgctgccc gcgacctgat
3901 ctgcgcccag aagttcaatg gcctgaccgt gctgccacca ctgctgaccg atgagatgat
3961 cgctcagtac acctccgccc tgctggccgg aaccatcacc agcggatgga ccttcggcgc
4021 tggagccgcc ctgcagatcc ccttcgccat gcagatggcc taccgcttca acggcatcgg
4081 cgtgacccag aatgtgctgt acgagaacca gaagctgatc gccaatcagt tcaactccgc
4141 catcggcaag atccaggact ccctgtccag caccgccagc gccctgggca agctgcagga
4201 tgtggtgaat cagaacgccc aggccctgaa taccctggtg aagcagctgt ccagcaactt
4261 cggcgccatc tccagcgtgc tgaatgatat cctgagccgc ctggacaagg tggaggctga
4321 ggtgcagatc gataggctga tcaccggccg cctgcagtcc ctgcagacct acgtgaccca
4381 gcagctgatc agggctgctg agatcagggc cagcgccaat ctggctgcta ccaagatgtc
4441 cgagtgcgtg ctgggacaga gcaagagggt ggacttctgc ggcaagggct accacctgat
4501 gtccttccca cagagcgccc cacacggagt ggtgttcctg cacgtgacct acgtgccagc
4561 ccaggagaag aacttcacca ccgctccagc tatctgccac gatggcaagg ctcacttccc
4621 acgcgagggc gtgttcgtgt ccaacggcac ccactggttc gtgacccagc gcaatttcta
4681 cgagccccag atcatcacca ccgacaatac cttcgtgagc ggcaactgcg acgtggtcat
4741 cggaatcgtg aacaataccg tgtacgatcc cctgcagcca gagctggact ccttcaagga
4801 ggagctggat aagtacttca agaatcacac cagccccgac gtggatctgg gcgacatctc
4861 cggcatcaat gccagcgtgg tgaacatcca gaaggagatc gaccgcctga acgaggtggc
4921 caagaatctg aacgagtccc tgatcgatct gcaggagctg ggcaagtacg agcagtacat
4981 caagtggcca tggtacatct ggctgggctt catcgccggc ctgatcgcca tcgtgatggt
5041 gaccatcatg ctgtgctgca tgacctcctg ctgcagctgc ctgaagggct gctgctcctg
5101 cggcagctgc tgcaagttcg atgaggacga tagcgagccc gtgctgaagg gcgtcaaact
5161 gcactataca tagctcgaca tataaggatc cactcgacga ctcgacaatc aacctctgga
5221 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
5281 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
5341 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
5401 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
5461 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
5521 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
5581 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
5641 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
5701 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg catcttcgcc ttcgccctca
5761 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattcga gctcggtacc
5821 ttgattctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
5881 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
5941 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
6001 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
6061 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagtcaa
6121 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
6181 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
6241 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
6301 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
6361 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
6421 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
6481 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
6541 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
6601 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
6661 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
6721 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
6781 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
6841 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
6901 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
6961 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
7021 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
7081 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
7141 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
7201 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
7261 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
7321 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
7381 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
7441 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
7501 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
7561 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
7621 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
7681 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
7741 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
7801 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
7861 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
7921 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
7981 acctccacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
8041 tgatgccgca tagtt
//
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