| 质粒名称: | CoV2-N-WT-Hu1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 177937 |
| 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: | 6658 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 Nucleocapsid (original Wuhan Hu1 sequence) |
| 物种来源: | Other |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | CoV2-N-WT-Hu1 was a gift from Jennifer Doudna (Addgene plasmid # 177937 ; http://n2t.net/addgene:177937 ; RRID:Addgene_177937) |
| 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-N-WT-Hu1 6658 bp ds-DNA circular SYN 16-NOV-2021
DEFINITION Transient mammalian expression of codon optimized SARS-CoV-2
Nucleocapsid (original Wuhan Hu1 sequence). Used for generating RNA
packaging virus-like particles..
ACCESSION .
VERSION .
KEYWORDS CoV2-N-WT-Hu1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6658)
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 6658)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 16, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6658
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 72..451
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 452..655
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 606..626
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 700..719
/label=T7
/note="T7 promoter, forward primer"
promoter 700..718
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2086..2103)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2092..2316
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2362..2790
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2449..2468)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2659..2680
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2799..2819)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2804..3133
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2984..3119
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3046..3065
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3200..3994
/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(3254..3273)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3864..3883
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4168..4289
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4205..4224)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4259..4278
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4338..4354)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4338..4354)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4351..4373)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4362..4378
/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(4386..4416)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4431..4452
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4569..4586)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4740..5328)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4820..4839)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5499..6359)
/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 6122..6141
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6360..6464)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6539..6558)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
61 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
121 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
181 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
241 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
301 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
361 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
421 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
481 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
541 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
601 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
661 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
721 gacccaagct ggctagcctt ccatttcagg tgtcgtgagg atctatttcc ggtgaattcg
781 ccgccaccat gagcgataac ggcccccaaa accaacggaa tgcccctcgg ataacatttg
841 gtggccctag tgattccaca ggaagtaacc aaaacggaga acggtctgga gcccgctcta
901 aacagaggcg accacaggga ttgccaaata acacagcgtc atggtttaca gccctgactc
961 aacatggaaa ggaggacttg aaatttccac gaggacaggg tgtaccaata aacacaaaca
1021 gtagtccaga cgatcagatc gggtattata gaagggccac gagacgcatc agaggtggag
1081 atggtaaaat gaaagatctc agtccgcgct ggtactttta ttatttggga accggcccag
1141 aagcgggcct cccatatgga gccaataagg acgggattat atgggtggct acagagggag
1201 ccctcaatac gccaaaagat cacatcggaa caagaaaccc tgctaataat gccgcaatcg
1261 tgctgcagtt gcctcagggg acgacgcttc ctaaaggctt ttacgcagaa ggatcacgcg
1321 gcggcagcca agcatcctca aggtctagtt ccagaagtcg aaacagctcc agaaactcca
1381 caccagggtc cagtaggggc acaagtccgg cgcggatggc gggcaacggc ggagacgccg
1441 cactcgctct gttgctcctg gaccggctca accaacttga atccaagatg agtggtaagg
1501 gacagcagca acaaggtcaa accgtaacca agaaaagcgc tgcagaagct tctaaaaaac
1561 ctcgacaaaa acggaccgct acgaaggcat ataacgttac tcaagccttc gggagacgag
1621 ggccggagca aacccagggg aattttggag atcaggagct catccggcaa gggacagatt
1681 acaaacattg gcctcaaatc gctcagttcg cgccgtccgc aagtgctttt ttcggaatgt
1741 ctcgcatcgg tatggaagta acaccatccg ggacatggct cacttatacc ggcgccatca
1801 agctggacga caaagatcca aatttcaaag atcaggtgat tctcttgaat aaacacatag
1861 acgcgtataa gacgtttcca cccaccgaac ccaagaagga caaaaagaaa aaagctgatg
1921 aaacacaagc gttgccccag cgacagaaga agcagcaaac cgtcaccctc ctgccagccg
1981 ctgatctcga tgacttctcc aagcaacttc aacagtccat gagctcagcc gattctactc
2041 aggcgtaact cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
2101 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
2161 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
2221 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
2281 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
2341 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
2401 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
2461 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
2521 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
2581 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
2641 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
2701 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
2761 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
2821 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
2881 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
2941 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
3001 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
3061 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
3121 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
3181 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
3241 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
3301 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
3361 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
3421 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
3481 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
3541 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
3601 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
3661 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
3721 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
3781 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
3841 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
3901 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
3961 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
4021 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
4081 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
4141 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
4201 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
4261 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
4321 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
4381 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
4441 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
4501 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
4561 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
4621 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
4681 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
4741 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
4801 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
4861 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
4921 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
4981 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
5041 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
5101 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
5161 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
5221 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
5281 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
5341 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
5401 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
5461 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
5521 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
5581 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
5641 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
5701 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
5761 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
5821 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
5881 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
5941 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
6001 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
6061 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
6121 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
6181 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
6241 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
6301 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
6361 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
6421 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
6481 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
6541 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
6601 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaa
//
