| 质粒名称: | pCAG-FAST-p10 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89152 |
| 存储实验室: | Takeshi Kobayashi |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGGS |
| 空载体大小: | 5713 bp |
| 完整质粒大小: | 6001 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 嘌呤霉素 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
|
插入基因信息 |
|
| 插入基因名称: | NBV p10-FAST |
| 物种来源: | 内尔森湾正呼肠孤病毒 |
| 插入大小: | 288 bp |
| GenBank编号 | AB521793 |
| 启动子: | CAG promotor |
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | CTAGAGCCTCTGCTAACCATGTTC |
| 3’测序引物 | TCAGTGGTATTTGTGAGCCAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-FAST-p10 was a gift from Takeshi Kobayashi (Addgene plasmid # 89152 ; http://n2t.net/addgene:89152 ; RRID:Addgene_89152) |
| 参考文献部分: | Entirely plasmid-based reverse genetics system for rotaviruses. Kanai Y, Komoto S, Kawagishi T, Nouda R, Nagasawa N, Onishi M, Matsuura Y, Taniguchi K, Kobayashi T. Proc Natl Acad Sci U S A. 2017 Jan 30. pii: 201618424. doi: 10.1073/pnas.1618424114. 10.1073/pnas.1618424114 PubMed 28137864 |
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 pCAG-FAST-p10 6001 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express p10-FAST in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pCAG-FAST-p10
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6001)
AUTHORS Kanai Y, Komoto S, Kawagishi T, Nouda R, Nagasawa N, Onishi M,
Matsuura Y, Taniguchi K, Kobayashi T
TITLE Entirely plasmid-based reverse genetics system for rotaviruses.
JOURNAL Proc Natl Acad Sci U S A. 2017 Jan 30. pii: 201618424. doi:
10.1073/pnas.1618424114.
PUBMED 28137864
REFERENCE 2 (bases 1 to 6001)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6001
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 382..657
/label=chicken beta-actin promoter
intron 658..1672
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1596..1618
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1680..1699
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
primer_bind complement(2146..2165)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2211..2266
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2265..2284)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(2618..2638)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
primer_bind 2865..2884
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3045..3644
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3045..3064)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3541..3561
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 3874..4008
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3911..3930)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3965..3984
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4076..4093)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4247..4835)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4327..4346)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5006..5866)
/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 5629..5648
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5867..5971)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
481 tgggggcggg gggggggggg gggggcgcca ggcggggcgg ggcggggcga ggggcggggc
541 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
601 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
661 gtcgctgcgt tgccttcgcc ccgtgccccg ctccgcgccg cctcgcgccg cccgccccgg
721 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
781 tgtaattagc gcttggttta atgacggctc gtttcttttc tgtggctgcg tgaaagcctt
841 aaagggctcc gggagggccc tttgtgcggg ggggagcggc tcggggggtg cgtgcgtgtg
901 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
961 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
1021 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
1081 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
1141 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
1201 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
1261 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
1321 tcgaggcgcg gcgagccgca accattgcct tttatggtaa tcgtgcgaga gggcgcaagg
1381 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
1501 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
1561 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
1621 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
1681 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattaatta agtttaaacg
1741 gcgcgccgat atcttaagat ctatgagcgg tgattgcgct ggtctggttt cagtgtttgg
1801 gagtgttcac tgtcagtcgt ctaagaataa agccggtggg gatctgcaag cgacttctgt
1861 tctaaccaca tattggccgc atcttgcgat tggtggcagt atcatcttga tcatcctcct
1921 gctgggtcta ttttactgtt gttatctcaa gtggaagacg tcacatattc gtcgtactta
1981 tcacaaggag ttggtggcgc taactcgcgg ctatgtcagg ccgatcccgg cggacgttac
2041 cagtgtctga agatcttcga attcactcct caggtgcagg ctgcctatca gaaggtggtg
2101 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atcgatcttt ttccctctgc
2161 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
2221 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
2281 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
2341 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
2401 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
2461 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
2521 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
2581 cttatggaga tccctcgacc tgcagcccaa gctctgtgga atgtgtgtca gttagggtgt
2641 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
2701 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
2761 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
2821 cccagtttcc gcccttctcc gccccatggc tgactaattt tttttattta tgcagaggcc
2881 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
2941 ggcttttgca aaaagcttgc atgcctgcag gtcggccgcc acgaccggtg ccgccaccat
3001 cccctgaccc acgcccctga cccctcacaa ggagacgacc ttccatgacc gagtacaagc
3061 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
3121 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg
3181 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
3241 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
3301 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
3361 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
3421 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
3481 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
3541 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
3601 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc
3661 cgcagcgccc gaccgaaagg agcgcacgac cccatggctc cgaccgaagc cacccggggc
3721 ggccccgccg accccgcacc cgcccccgag gcccaccgac tctagaggat cataatcagc
3781 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
3841 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
3901 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
3961 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat
4021 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
4081 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
4141 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
4201 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
4261 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
4321 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
4381 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
4441 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
4501 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
4561 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
4621 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
4681 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
4741 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
4801 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
4861 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
4921 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
4981 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
5041 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
5101 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
5161 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
5221 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
5281 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
5341 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
5401 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
5461 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
5521 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
5581 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
5641 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
5701 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
5761 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
5821 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
5881 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
5941 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt
6001 c
//
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