pSFV1 vector (V001651) Gene synthesis in pSFV1 backbone

COA reports Sequencing Results MSDS Download GenBank File(.gb)

Price Information

Cat No.	Plasmid Name	Availability	Buy one, get one free! (?)
V001651	pSFV1	In stock, instant shipping

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

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.

Vector Name:: pSFV1

Antibiotic Resistance:: Ampicillin

Length:: 11029 bp

Type:: Mammalian Expression Vectors

Replication origin:: ori

Growth Strain(s):: Stbl3

Growth Temperature:: 37℃

pSFV1 vector Map

Copy Sequence View Map

Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5. Store the plasmid at -20 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

References

Pinnock EC, Jovanovic K, Pinto MG, Ferreira E, Dias Bda C, Penny C, Knackmuss S, Reusch U, Little M, Schatzl HM, Weiss SF. LRP/LR Antibody Mediated Rescuing of Amyloid-β-Induced Cytotoxicity is Dependent on PrPc in Alzheimer's Disease. J Alzheimers Dis. 2016;49(3):645-57.

pSFV1 vector Sequence

LOCUS       40924_39818       11029 bp DNA     circular SYN 24-NOV-2023
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11029)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             87..7382
                     /codon_start=1
                     /product="Nonstructural polyprotein from Semliki Forest
                     virus"
                     /label=Nonstructural polyprotein
                     /translation="MAAKVHVDIEADSPFIKSLQKAFPSFEVESLQVTPNDHANARAFS
                     HLATKLIEQETDKDTLILDIGSAPSRRMMSTHKYHCVCPMRSAEDPERLDSYAKKLAAA
                     SGKVLDREIAGKITDLQTVMATPDAESPTFCLHTDVTCRTAAEVAVYQDVYAVHAPTSL
                     YHQAMKGVRTAYWIGFDTTPFMFDALAGAYPTYATNWADEQVLQARNIGLCAASLTEGR
                     LGKLSILRKKQLKPCDTVMFSVGSTLYTESRKLLRSWHLPSVFHLKGKQSFTCRCDTIV
                     SCEGYVVKKITMCPGLYGKTVGYAVTYHAEGFLVCKTTDTVKGERVSFPVCTYVPSTIC
                     DQMTGILATDVTPEDAQKLLVGLNQRIVVNGRTQRNTNTMKNYLLPIVAVAFSKWAREY
                     KADLDDEKPLGVRERSLTCCCLWAFKTRKMHTMYKKPDTQTIVKVPSEFNSFVIPSLWS
                     TGLAIPVRSRIKMLLAKKTKRELIPVLDASSARDAEQEEKERLEAELTREALPPLVPIA
                     PAETGVVDVDVEELEYHAGAGVVETPRSALKVTAQPNDVLLGNYVVLSPQTVLKSSKLA
                     PVHPLAEQVKIITHNGRAGGYQVDGYDGRVLLPCGSAIPVPEFQALSESATMVYNEREF
                     VNRKLYHIAVHGPSLNTDEENYEKVRAERTDAEYVFDVDKKCCVKREEASGLVLVGELT
                     NPPFHEFAYEGLKIRPSAPYKTTVVGVFGVPGSGKSAIIKSLVTKHDLVTSGKKENCQE
                     IVNDVKKHRGKGTSRENSDSILLNGCRRAVDILYVDEAFACHSGTLLALIALVKPRSKV
                     VLCGDPKQCGFFNMMQLKVNFNHNICTEVCHKSISRRCTRPVTAIVSTLHYGGKMRTTN
                     PCNKPIIIDTTGQTKPKPGDIVLTCFRGWAKQLQLDYRGHEVMTAAASQGLTRKGVYAV
                     RQKVNENPLYAPASEHVNVLLTRTEDRLVWKTLAGDPWIKVLSNIPQGNFTATLEEWQE
                     EHDKIMKVIEGPAAPVDAFQNKANVCWAKSLVPVLDTAGIRLTAEEWSTIITAFKEDRA
                     YSPVVALNEICTKYYGVDLDSGLFSAPKVSLYYENNHWDNRPGGRMYGFNAATAARLEA
                     RHTFLKGQWHTGKQAVIAERKIQPLSVLDNVIPINRRLPHALVAEYKTVKGSRVEWLVN
                     KVRGYHVLLVSEYNLALPRRRVTWLSPLNVTGADRCYDLSLGLPADAGRFDLVFVNIHT
                     EFRIHHYQQCVDHAMKLQMLGGDALRLLKPGGILMRAYGYADKISEAVVSSLSRKFSSA
                     RVLRPDCVTSNTEVFLLFSNFDNGKRPSTLHQMNTKLSAVYAGEAMHTAGCAPSYRVKR
                     ADIATCTEAAVVNAANARGTVGDGVCRAVAKKWPSAFKGAATPVGTIKTVMCGSYPVIH
                     AVAPNFSATTEAEGDRELAAVYRAVAAEVNRLSLSSVAIPLLSTGVFSGGRDRLQQSLN
                     HLFTAMDATDADVTIYCRDKSWEKKIQEAIDMRTAVELLNDDVELTTDLVRVHPDSSLV
                     GRKGYSTTDGSLYSYFEGTKFNQAAIDMAEILTLWPRLQEANEQICLYALGETMDNIRS
                     KCPVNDSDSSTPPRTVPCLCRYAMTAERIARLRSHQVKSMVVCSSFPLPKYHVDGVQKV
                     KCEKVLLFDPTVPSVVSPRKYAASTTDHSDRSLRGFDLDWTTDSSSTASDTMSLPSLQS
                     CDIDSIYEPMAPIVVTADVHPEPAGIADLAADVHPEPADHVDLENPIPPPRPKRAAYLA
                     SRAAERPVPAPRKPTPAPRTAFRNKLPLTFGDFDEHEVDALASGITFGDFDDVLRLGRA
                     GAYIFSSDTGSGHLQQKSVRQHNLQCAQLDAVQEEKMYPPKLDTEREKLLLLKMQMHPS
                     EANKSRYQSRKVENMKATVVDRLTSGARLYTGADVGRIPTYAVRYPRPVYSPTVIERFS
                     SPDVAIAACNEYLSRNYPTVASYQITDEYDAYLDMVDGSDSCLDRATFCPAKLRCYPKH
                     HAYHQPTVRSAVPSPFQNTLQNVLAAATKRNCNVTQMRELPTMDSAVFNVECFKRYACS
                     GEYWEEYAKQPIRITTENITTYVTKLKGPKAAALFAKTHNLVPLQEVPMDRFTVDMKRD
                     VKVTPGTKHTEERPKVQVIQAAEPLATAYLCGIHRELVRRLNAVLRPNVHTLFDMSAED
                     FDAIIASHFHPGDPVLETDIASFDKSQDDSLALTGLMILEDLGVDQYLLDLIEAAFGEI
                     SSCHLPTGTRFKFGAMMKSGMFLTLFINTVLNITIASRVLEQRLTDSACAAFIGDDNIV
                     HGVISDKLMAERCASWVNMEVKIIDAVMGEKPPYFCGGFIVFDSVTQTACRVSDPLKRL
                     FKLGKPLTAEDKQDEDRRRALSDEVSKWFRTGLGAELEVALTSRYEVEGCKSILIAMAT
                     LARDIKAFKKLRGPVIHLYGGPRLVR"
     rep_origin      complement(8499..9087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9261..10118)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(10119..10223)
                     /label=AmpR promoter