pBG210 plasmid Sindbis virus辛德毕斯病毒质粒载体 BioVector NTCC质粒载体菌种细胞基因保藏中心

pBG210 plasmid Sindbis virus辛德毕斯病毒质粒载体 BioVector NTCC质粒载体菌种细胞基因保藏中心
pBG210 Recombinant Sindbis Virus Expression Plasmid – Gateway CompatibleRecombinant plasmid pBG210 containing Sindbis virus (SINV) allows for fast gene of interest addition with Gateway recombination.Highlights:    Contains a Gateway recombination cassette to allow for efficient addition of genes of interest behind 3’ subgenomic promoterThe entire Sindbis virus genome was inserted into the pcDNA3.1 Zeo+ expression vector such that the transcription start site of the CMV promoter overlaps with the first viral nucleotide of the viral genome. A Hepatitis D virus (HDV) ribozyme was added to the 3' end of the viral genomic 35bp polyA sequence. A Gateway recombination cassette is present behind the 3' subgenomic promoter to allow genes of interest in pDONR plasmids to be easily inserted into the replicon. Transfection of Gateway recombined versions of construct into mammalian cells produces a self-replicating Sindbis virus genome that expresses the genes of interest that will be expressed only when the virus genome is replicated. Transfection of Gateway recombined variants of this construct into mammalian cells produces an infectious self-replicating Sindbis virus genome that can express genes of interest in vitro or in vivo.
Product Type:  PlasmidGene/insert name:  Sindbis virus repliconOrganism:  Sindbis Virus Strain TE3'2J (AR339)Antibiotic Resistance:  Amp/chloramphenciolGrow in E. coli at 37 C:  DB3.1 or similar ccdB Survival E. Coli Cells prior to Gateway Recombination, DH5-alpha or XL10-Gold E-coli after Gateway Recombination, 37CCloning Site 5':  MluICloning Site 3':  XhoIInsert Size:  12,169 bpVector Backbone and Size:  pcDNA3.1 Zeo+High or low copy:  HighStorage:  Room Temperature (Dried), -20C (liquid)Shipped:  Room Temperature
Plasmid constructs. The full-length double subgenomic Sindbis genome TE3'2J was engineered behind a cytomegalovirus promoter (CMV), and a 3' hepatitis delta virus ribozyme (HDV) was added after a 35 base adenine repeat (A35) to cleave the viral RNA from the over length transcript. pBG167 possess a unique XbaI site after the second SGP, and is analogous to pTE3'2J [17]. pBG218 is pBG167 with GFP inserted at XbaI site via ligation between the unique XbaI site in pBG167 and engineered NheI sites flanking the GFP ORF. pBG210 and pBG211 have the attR1/attR2 Gateway Recombination Cassettes ligated into the pBG167 XbaI site in forward and reverse orientation, respectively. pBG212 is the product of a LR Clonase reaction between pBG210 and pBG440 and places GFP in the sense orientation. pBG213 is a recombination between pBG211 and pBG440, and places GFP in the reverse orientation. pBG451 is the product of a recombination between pBG210 and pBG403, and places Renilla luciferase in the sense orientation. pBG452 is the product of a recombination between pBG210 and pBG344 and places mCherry in the sense orientation.
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