The pSFV1 Eukaryotic Expression Vector is a novel DNA expression system based on the Semliki Forest Virus (SFV) replicon which has a broad host range and a high level of expression efficiency (1). In contrast to other protein expression systems, the SFV expression system has several advantages, including direct cloning into the vector and proper glycosylation of the recombinant protein. The SFV expression system is easy to use. The DNA of interest is cloned into the pSFV1 plasmid vector that serves as a template for in vitro synthesis of recombinant RNA, and the RNA is then transfected into tissue culture cells with DMRIE-C Reagent, Lipofectin Reagent or by electroporation. The recombinant RNA in the cell drives its own replication and capping, resulting in production of large amounts of heterologous protein while inhibiting host protein synthesis. High transfection efficiencies and expression of heterologous proteins have been achieved by transfection of pSFV1 in vitro transcribed RNA into CHO-K1 and BHK-21 cells (2). The plasmid pSFV3- lacZ contains the E. coli #-galactosidase gene, lacZ, and is included as a control plasmid to optimize transfection conditions.
| 载体基本信息 | |||
| 载体名称 | pSFV1 | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 11033 bp | 载体类型 | Mammalian Expression Vectors |
pSFV1 载体载体图谱
pSFV1 载体载体序列
LOCUS Exported 11033 bp ds-DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11033)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..11033
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(8503..9091)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9262..10122)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(10123..10227)
/gene="bla"
/note="AmpR promoter"
promoter 11017..2
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
