| 质粒名称: | SARS-CoV-2 S HexaPro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 154754 |
| 存储实验室: | Jason McLellan |
|
载体骨架基本信息 |
|
| 载体名称: | pαH |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4746 bp |
| 完整质粒大小: | 8370 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Spike (HexaPro variant) |
| 物种来源: | Severe acute respiratory syndrome coronavirus 2 |
| 插入大小: | 3624 bp |
| 基因突变: | Ectodomain only (AAs 1-1208); 682-685 (furin site) replaced with 'GSAS'; F817P, A892P, A899P, A942P, K986P, V987P |
| GenBank编号 | MN908947 |
| 启动子 | -- |
| 标签/融合蛋白: |
HRV 3C cleavage site (before tags)（主干上的C末端）
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: | SARS-CoV-2 S HexaPro was a gift from Jason McLellan (Addgene plasmid # 154754 ; http://n2t.net/addgene:154754 ; RRID:Addgene_154754) |
| 参考文献部分: | Structure-based design of prefusion-stabilized SARS-CoV-2 spikes. Hsieh CL, Goldsmith JA, Schaub JM, DiVenere AM, Kuo HC, Javanmardi K, Le KC, Wrapp D, Lee AG, Liu Y, Chou CW, Byrne PO, Hjorth CK, Johnson NV, Ludes-Meyers J, Nguyen AW, Park J, Wang N, Amengor D, Lavinder JJ, Ippolito GC, Maynard JA, Finkelstein IJ, McLellan JS. Science. 2020 Sep 18;369(6510):1501-1505. doi: 10.1126/science.abd0826. Epub 2020 Jul 23. 10.1126/science.abd0826 PubMed 32703906 |
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 SARS-CoV-2 S HexaPro 8376 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression vector for expression of the SARS-CoV-2 spike
HexaPro variant.
ACCESSION .
VERSION .
KEYWORDS SARS-CoV-2 S HexaPro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8376)
AUTHORS Hsieh CL, Goldsmith JA, Schaub JM, DiVenere AM, Kuo HC, Javanmardi
K, Le KC, Wrapp D, Lee AG, Liu Y, Chou CW, Byrne PO, Hjorth CK,
Johnson NV, Ludes-Meyers J, Nguyen AW, Park J, Wang N, Amengor D,
Lavinder JJ, Ippolito GC, Maynard JA, Finkelstein IJ, McLellan JS
TITLE Structure-based design of prefusion-stabilized SARS-CoV-2 spikes.
JOURNAL Science. 2020 Sep 18;369(6510):1501-1505. doi:
10.1126/science.abd0826. Epub 2020 Jul 23.
PUBMED 32703906
REFERENCE 2 (bases 1 to 8376)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8376
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 662..1678
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1602..1624
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
CDS 5486..5509
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 5513..5536
/codon_start=1
/product="8xHis affinity tag"
/label=8xHis
/translation="HHHHHHHH"
CDS 5543..5566
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 5606..5629
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
primer_bind complement(5731..5750)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 5796..5851
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(5850..5869)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(6212..6228)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6212..6228)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6225..6247)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6236..6252
/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(6260..6290)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6305..6326
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6455..6472)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6626..7214)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6706..6725)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7385..8245)
/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 8008..8027
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8246..8350)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggggcgc gccrgggggg gggggggggg gggggggggg
541 ggggggggsg rggsggagag gkgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
1261 ggggccgcct cgggccgggg rgggctcggg ggaggggcgc ggcggccccc ggagcgccgg
1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
1621 gaccggcggc tctagcgcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tgcggccgtc
1741 tcaggccgag ttcggtaccg ccaccatgtt cgtgttcctg gtgctcctgc ctctggtgag
1801 cagccagtgc gtgaacctga ccacccgaac ccagctccca ccagcctaca ccaacagctt
1861 tacacggggc gtgtactacc ctgacaaggt gttcagatct agcgtcctgc acagcactca
1921 ggacctcttc ctgccgttct tcagcaacgt gacatggttc cacgccatcc acgtgagcgg
1981 cacaaacgga accaagcggt ttgataaccc cgtcctgcca ttcaatgatg gagtttactt
2041 cgccagtacc gagaagagta acatcatccg gggctggatc ttcggcacca ccctggatag
2101 caaaacacag agcctcctga tcgtgaacaa tgccacgaac gtcgtgatca aggtgtgcga
2161 gttccagttt tgcaatgatc ctttcctggg tgtgtactac cacaagaaca acaagagctg
2221 gatggaaagc gagttcagag tctacagcag cgccaacaac tgcacattcg agtacgtctc
2281 tcagcctttt ctgatggacc ttgaggggaa acaaggcaac ttcaagaacc tgagagaatt
2341 cgtgttcaag aacatcgacg gctacttcaa aatctactcc aagcacacac ccatcaacct
2401 ggtccgggac ctccctcagg gcttcagcgc cctggaaccc ctggtcgacc tgcccatagg
2461 catcaacata acgcggttcc aaaccctgct ggccctgcat agatcctacc tgactcctgg
2521 cgacagcagc agcggatgga ccgccggagc tgcagcctac tatgtgggct acctgcaacc
2581 tagaaccttc ctgctgaagt acaacgagaa cggcacaatc acagacgccg tcgactgcgc
2641 cctggaccct ctctctgaga caaagtgcac cctgaagtcc ttcaccgtgg aaaagggcat
2701 ctaccagacc agcaacttcc gggtgcagcc tacagagagc atcgtgcgat ttccaaacat
2761 taccaacctc tgccccttcg gcgaggtgtt taacgccaca agatttgcct ccgtttacgc
2821 ctggaataga aagagaatca gcaattgtgt ggccgactac tccgtgctgt ataacagcgc
2881 ctctttcagc accttcaagt gctacggcgt ttccccaaca aagctgaatg acctgtgctt
2941 caccaacgtg tacgccgact ccttcgtaat tagaggcgat gaggtgcggc agatcgcacc
3001 aggccagacc ggtaagatcg ctgactacaa ctataagctg cctgatgatt ttacaggctg
3061 cgtgatcgcc tggaactcta acaacctgga tagcaaggtg ggcggcaact acaactacct
3121 gtaccggctg tttcgcaagt ctaacctgaa acctttcgag agagacatct ccacagagat
3181 ctaccaggcc ggttctacac cttgtaacgg ggtggaaggc ttcaactgtt acttccctct
3241 gcaaagctac ggcttccagc ctaccaatgg agtcggctac cagccatacc gggtggtcgt
3301 gctgtccttc gagttactcc acgcccccgc caccgtctgc ggtcctaaga agtccaccaa
3361 tctggttaag aacaaatgcg tgaacttcaa cttcaacggc ctgaccggga ccggcgtgct
3421 gaccgaaagc aacaaaaagt tcctcccctt ccagcagttc ggccgtgata tcgctgacac
3481 cacagatgcc gtcagagatc cacagaccct ggaaatcctg gatattacac cctgctcctt
3541 cggaggagtt tctgtgatca cccccgggac caataccagc aaccaggtgg ctgtgctgta
3601 ccaagatgtt aactgcaccg aggttcctgt ggccatccac gccgatcagc tgacacctac
3661 ttggagagtg tactccactg gctccaatgt gttccagacc agggccggat gtctgatcgg
3721 cgccgagcac gtgaataaca gttacgagtg cgacatccct atcggcgccg gcatctgtgc
3781 cagctaccag acccagacaa acagccctgg gtctgcttcc tctgtagcta gccagagcat
3841 catcgcctac accatgagcc tgggcgcaga gaacagcgtg gcctattcca acaactctat
3901 cgccattccc accaacttta caattagcgt cacaacagag atcctgcccg tgagcatgac
3961 caagaccagc gtggactgta caatgtacat ctgtggcgac agcactgaat gcagcaacct
4021 gctgctgcaa tacggctcct tttgcaccca actgaaccgg gcgctgaccg gaatcgccgt
4081 ggaacaggac aaaaataccc aggaggtgtt cgcccaagtg aagcagatct acaagacccc
4141 acctatcaag gacttcggcg gctttaactt tagccagatt ctccctgatc cttctaagcc
4201 tagcaagcgg agccctatcg aggatctgct gttcaacaag gtcaccctgg ccgatgccgg
4261 ctttatcaaa cagtatggcg attgcctggg cgacatagcc gccagagatc tgatctgcgc
4321 ccagaaattc aacggcctga cagttctccc acctctgctg accgacgaga tgatcgctca
4381 gtacacctct gccctgctgg ctggcaccat cacatctggg tggacatttg gcgccggccc
4441 cgccctgcag atcccctttc ccatgcagat ggcctataga ttcaacggaa tcggcgtgac
4501 ccagaacgtg ctgtatgaaa accagaagct gatcgctaac cagttcaatt ctgccatcgg
4561 caagatccag gactccctct cctctacccc cagcgccctg ggcaaactgc aggacgtggt
4621 gaatcagaac gcccaagccc tgaacaccct ggtgaagcag ctcagcagca attttggcgc
4681 catcagctct gtgctgaacg atatcctgtc tagactggac cctccagaag ccgaagtcca
4741 gatcgataga ctgatcacag gcagactgca gtccctgcaa acctacgtga cccaacagct
4801 gatcagggcc gctgaaataa gagccagcgc caatctcgcc gctaccaaga tgtccgagtg
4861 tgtgctggga cagtctaaac gcgttgactt ctgcggcaaa ggctatcacc tgatgagctt
4921 cccccagagc gcgccgcacg gcgtggtgtt cctgcatgtg acatacgtgc ctgcccaaga
4981 gaagaatttc acaaccgccc ctgccatctg ccacgacggc aaggcccact tcccaagaga
5041 gggcgttttc gtttccaatg gcacacactg gttcgtgaca caaagaaact tctacgaacc
5101 ccagattatc accaccgaca acaccttcgt gagtggcaat tgtgacgtgg tcatcggaat
5161 cgtgaacaac acagtgtacg accctctgca acctgagctg gactctttta aggaagagct
5221 ggacaagtac tttaaaaacc acaccagccc cgatgtggac ctgggcgaca tcagtggcat
5281 taacgccagc gtggtgaaca tccaaaagga aatcgacaga ctgaacgagg tggccaagaa
5341 cctgaacgag tccctgatcg acctgcagga gctcggcaaa tacgagcagg gatccggata
5401 catccccgag gcccccagag atggccaggc ctacgtgcgg aaggacggcg agtgggtact
5461 gctgagcaca ttcctgggca gatccctgga ggtgctgttc cagggcccag gccatcacca
5521 ccatcaccac catcatagcg cctggtccca cccccagttc gagaagggcg gcggtagtgg
5581 agggggcgga tctggcggct cagcttggag ccacccccag ttcgaaaagt gataatgact
5641 cgagcgataa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
5701 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
5761 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
5821 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
5881 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
5941 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
6001 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
6061 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
6121 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
6181 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
6241 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
6301 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
6361 gaaacctgtc gtgccagcgg atcgatccgc tgcattaatg aatcggccaa cgcgcgggga
6421 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
6481 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
6541 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
6601 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
6661 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
6721 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
6781 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
6841 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
6901 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
6961 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
7021 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
7081 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
7141 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
7201 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
7261 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
7321 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
7381 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
7441 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
7501 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
7561 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
7621 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
7681 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
7741 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
7801 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
7861 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
7921 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
7981 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
8041 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
8101 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
8161 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
8221 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
8281 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
8341 gggttccgcg cacatttccc cgaaaagtgc cacctg
//
