| 质粒名称: | pCAGGS_NP_EBOV |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 103049 |
| 存储实验室: | Elke Mühlberger |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGGs |
| 载体出品公司: | Clontech |
| 空载体大小: | 7132 bp |
| 完整质粒大小: | 7132bp |
| 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: | NP gene of Ebola virus、nucleoprotein gene of Ebola virus |
| 物种来源: | Zaire ebolavirus, Mayinga |
| 插入大小: | 951 bp |
| 基因突变: | silent mutation (T to C) at position 928 of NP gene |
| 启动子: | CAG |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Sac 1 (unknown if destroyed) |
| 3’克隆位点: | Nhe 1 (unknown if destroyed) |
| 5’测序引物 | CAG-F GCAACGTGCTGGTTATTGTG |
|
文献引用方法 |
|
| 材料和方法部分: | pCAGGS_NP_EBOV was a gift from Elke Mühlberger (Addgene plasmid # 103049 ; http://n2t.net/addgene:103049 ; RRID:Addgene_103049) |
| 参考文献部分: | An RNA polymerase II-driven Ebola virus minigenome system as an advanced tool for antiviral drug screening. Nelson EV, Pacheco JR, Hume AJ, Cressey TN, Deflube LR, Ruedas JB, Connor JH, Ebihara H, Muhlberger E. Antiviral Res. 2017 Aug 12;146:21-27. doi: 10.1016/j.antiviral.2017.08.005. 10.1016/j.antiviral.2017.08.005 PubMed 28807685 |
买家导航
pCAGGS_NP_EBOV载体质粒基本信息
pCAGGS_NP_EBOV质粒图谱载体图谱和pCAGGS_NP_EBOV载体序列质粒序列多克隆位点信息
pCAGGS_NP_EBOV质粒载体简介
pCAGGS_NP_EBOV质粒序列载体序列
LOCUS pCAGGS_NP_EBOV 7132 bp DNA circular SYN 14-APR-2025
DEFINITION Encodes Ebola virus NP gene (Zaire ebolavirus, Mayinga) under
control of the CAG promoter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7132)
AUTHORS Nelson EV, Pacheco JR, Hume AJ, Cressey TN, Deflube LR, Ruedas JB,
Connor JH, Ebihara H, Muhlberger E
TITLE An RNA polymerase II-driven Ebola virus minigenome system as an
advanced tool for antiviral drug screening.
JOURNAL Antiviral Res. 2017 Aug 12;146:21-27. doi:
10.1016/j.antiviral.2017.08.005.
PUBMED 28807685
REFERENCE 2 (bases 1 to 7132)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Antiviral
Res."; date: "2017-08-12"; volume: "146:21-27. doi"; pages: "
10.1016/j.antiviral.2017.08.005"
COMMENT Sequence Label: pCAGGS_NP_EBOV
FEATURES Location/Qualifiers
source 1..7132
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
intron 662..1674
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1598..1620
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1682..1701
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 3335..3346
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(4155..4174)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 4220..4275
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4274..4293)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4635..4651)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4635..4651)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4648..4670)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4659..4675
/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(join(4683..4689,4690..4707,4708..4713))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4728..4749
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4808..5003
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4854..4989
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 5009..5143
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5046..5065)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5100..5119
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5211..5228)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5382..5970)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5462..5481)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(6141..6932,6933..7001))
/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 6764..6783
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7002..7106)
/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 tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcaag
1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
1621 ggcagctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcct cgagggtacc
1741 cccggggcgg ccgcgagctc actagtggat ccacaaaatt ccgagtatgg attctcgtcc
1801 tcagaaaatc tggatggcgc cgagtctcac tgaatctgac atggattacc acaagatctt
1861 gacagcaggt ctgtccgttc aacaggggat tgttcggcaa agagtcatcc cagtgtatca
1921 agtaaacaat cttgaagaaa tttgccaact tatcatacag gcctttgaag caggtgttga
1981 ttttcaagag agtgcggaca gtttccttct catgctttgt cttcatcatg cgtaccaggg
2041 agattacaaa cttttcttgg aaagtggcgc agtcaagtat ttggaagggc acgggttccg
2101 ttttgaagtc aagaagcgtg atggagtgaa gcgccttgag gaattgctgc cagcagtatc
2161 tagtggaaaa aacattaaga gaacacttgc tgccatgccg gaagaggaga caactgaagc
2221 taatgccggt cagtttctct ccttcgcaag tctattcctt ccgaaattgg tagtaggaga
2281 aaaggcttgc cttgagaagg ttcaaaggca aattcaagta catgcagagc aaggactgat
2341 acaatatcca acagcttggc aatcagtagg acacatgatg gtgattttcc gtttgatgcg
2401 aacaaatttt ctgatcaaat ttctcctaat acaccaaggg atgcacatgg ttgccgggca
2461 tgatgccaac gatgctgtga tttcaaattc agtggctcaa gctcgttttt caggcttatt
2521 gattgtcaaa acagtacttg atcatatcct acaaaagaca gaacgaggag ttcgtctcca
2581 tcctcttgca aggaccgcca aggtaaaaaa tgaggtgaac tcctttaagg ctgcactcag
2641 ctccctggcc aagcatggag agtatgctcc tttcgcccga cttttgaacc tttctggagt
2701 aaataatctt gagcatggtc ttttccctca actatcggca attgcactcg gagtcgccac
2761 agcacacggg agtaccctcg caggagtaaa tgttggagaa cagtatcaac aactcagaga
2821 ggctgccact gaggctgaga agcaactcca acaatatgca gagtctcgcg aacttgacca
2881 tcttggactt gatgatcagg aaaagaaaat tcttatgaac ttccatcaga aaaagaacga
2941 aatcagcttc cagcaaacaa acgctatggt aactctaaga aaagagcgcc tggccaagct
3001 gacagaagct atcactgctg cgtcactgcc caaaacaagt ggacattacg atgatgatga
3061 cgacattccc tttccaggac ccatcaatga tgacgacaat cctggccatc aagatgatga
3121 tccgactgac tcacaggata cgaccattcc cgatgtggtg gttgatcccg atgatggaag
3181 ctacggcgaa taccagagtt actcggaaaa cggcatgaat gcaccagatg acttggtcct
3241 attcgatcta gacgaggacg acgaggacac taagccagtg cctaatagat cgaccaaggg
3301 tggacaacag aagaacagtc aaaagggcca gcatatagag ggcagacaga cacaatccag
3361 gccaattcaa aatgtcccag gccctcacag aacaatccac cacgccagtg cgccactcac
3421 ggacaatgac agaagaaatg aaccctccgg ctcaaccagc cctcgcatgc tgacaccaat
3481 taacgaagag gcagacccac tggacgatgc cgacgacgag acgtctagcc ttccgccctt
3541 ggagtcagat gatgaagagc aggacaggga cggaacttcc aaccgcacac ccactgtcgc
3601 cccaccggct cccgtataca gagatcactc tgaaaagaaa gaactcccgc aagacgagca
3661 acaagatcag gaccacactc aagaggccag gaaccaggac agtgacaaca cccagtcaga
3721 acactctttt gaggagatgt atcgccacat tctaagatca caggggccat ttgatgctgt
3781 tttgtattat catatgatga aggatgagcc tgtagttttc agtaccagtg atggcaaaga
3841 gtacacgtat ccagactccc ttgaagagga atatccacca tggctcactg aaaaagaggc
3901 tatgaatgaa gagaatagat ttgttacatt ggatggtcaa caattttatt ggccggtgat
3961 gaatcacaag aataaattca tggcaatcct gcaacatcat cagtgaatga gcatggaaca
4021 atgggatgat tcaaccgaca aatagctaac attaagtagt caaggaacga aaacaggaag
4081 aatttttgat gtctaaggtg tgaattatta tcacaataaa agtgattctt atttttgaat
4141 ttaaagctag cagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
4201 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
4261 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
4321 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
4381 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
4441 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
4501 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
4561 tactcccagt catagctgtc cctcttctct tatgaagatc cctcgacctg cagcccaagc
4621 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
4681 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
4741 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
4801 cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
4861 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
4921 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
4981 ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa
5041 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
5101 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc
5161 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
5221 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
5281 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
5341 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
5401 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
5461 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
5521 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
5581 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
5641 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
5701 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
5761 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
5821 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
5881 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
5941 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
6001 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
6061 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
6121 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
6181 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
6241 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
6301 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
6361 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
6421 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
6481 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
6541 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
6601 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
6661 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
6721 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
6781 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
6841 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
6901 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
6961 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
7021 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
7081 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tg
//
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