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  • 產品名稱：pVision-RFP-NVector

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  • 產品廠商：Biovision
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簡單介紹：
pVision-RFP-NVector
詳情介紹：
Characteristics pVisionRFP-N is an eukaryotic (mammalian) expression vector encoding red fluorescent protein VisionRFP from sea anemone Entacmaea quadricolor. The vector allows to generate fusions to the VisionRFP N-terminus and to express VisionRFP fusions or VisionRFP alone in eukaryotic (mammalian) cells. pVisionRFP-N vector carries synthetic version of the VisionRFP gene which codon usage is humanized, i.e. optimized for high expression in mammalian cells. pVisionRFP-N vector backbone contains immediate early promoter of cytomegalovirus (P CMV IE ) for protein expression, SV40 origin for replication in mammalian cells, pUC origin of replication for propagation in E. coli, and f1 origin for single-stranded DNA production. SV40 early promoter provides neomycin resistance gene expression to select stably transfected eukaryotic cells using G418. Bacterial promoter (P) provides kanamycin resistance gene expression in E. coli. To increase VisionRFP mRNA translation efficiency, Kozak consensus translation initiation site is generated upstream of VisionRFP coding sequence. Multiple cloning site (MCS) is located between P CMV IE and VisionRFP coding sequence. MCS Nhel Bgl II Sac I Hind III EcoR I Sal I Kpn I Apa I GCT A.GC G.CT A.CCG.GAC.TC A.GAT. CT C. GAG. CTC. AAG.CTT. C GA.ATT. C TG.CA G. TCG.AC G.GTA. CC G.C GG. Eco47 III Xho I Pst I Sac II BamH I Age I RFP GCC.C G G.G AT.CC A.CCG.GT C.GCC.ACC. ATG.AGC.GAG SmaI/XmaI *
Application Notes Generation of VisionRFP-tagged fusions: A localization signal or a gene of interest should be cloned into MCS of the vector. It will be expressed as a fusion to VisionRFP N-terminus when inserted in the same reading frame as VisionRFP and no in-frame stop codons are present. VisionRFP-tagged fusions retain fluorescent properties of the native protein allowing fusion localization in vivo. Unmodified pVisionRFP-N vector will express VisionRFP when transfected into eukaryotic (mammalian) cells.
Note: Despite its dimeric structure, VisionRFP is still suitable for generation of fusions with proteins of interest.
Expression in Mammalian Cells: pVisionRFP-N can be transfected into mammalian cells by any known transfection method. CMV promoter provides strong, constitutive expression of VisionRFP or VisionRFP-tagged fusions in many cell types. If required, stable transformants can be selected using G418.
Propagation in E. coli: Suitable host strains: DH5alpha, HB101, and other general purpose strains. Single-stranded DNA production requires a host containing an F plasmid such as JM109 or XL1-Blue.
Selectable marker: plasmid confers resistance to kanamycin (30 μg/mL) to E. coli hosts.
E. coli replication origin: pUC
Copy number: approx. 500
Plasmid incompatibility group: pMB1/ColE1.
Restrictions For Research Use only
Storage -20 °C