OGS626
pSF-CMV-NEO-NH2-PPT-3XFLAG
(plasmid vector for molecular cloning)
Synonym(s):
cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, snapfast vector, vector
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About This Item
UNSPSC Code:
12352200
NACRES:
NA.85
Recommended Products
recombinant
expressed in human cells
tag
3X FLAG tagged
form
buffered aqueous solution
mol wt
size 5955 bp
bacteria selection
ampicillin
Origin of replication
pUC
Peptide cleavage
EKT
Peptide tag location
N-terminal
Promoter
Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian
shipped in
ambient
storage temp.
−20°C
General description
The pSF-CMV-NEO-NH2-PPT-3XFLAG expression vector is a 6 kb derivative of pSF-CMV-Amp for transient or stable expression of secreted N-terminal 3XFLAG fusion proteins in mammalian cells. This vector encodes three FLAG epitopes in tandem (DYKDHDG-DYKDHDI-DYKDDDDK), which results in increased sensitivity when using the anti-FLAG M2 antibody (catalogue number F3165). The third epitope includes the enterokinase recognition sequence, allowing cleavage of the 3XFLAG peptide from the purified fusion protein. The preprotrypsin (PPT) leader sequence precedes the 3XFLAG sequence and directs secretion of the fusion protein into the culture medium. pSF-CMV-NEO-NH2-PPT-3XFLAG is a shuttle vector, containing both E. coli and SV40 origins of replication, for propagation in bacterial and mammalian cells. Efficiency of replication and genomic integration is optimal when using an SV40 T antigen expressing mammalian host. The promoter regulatory region of the human cytomegalovirus drives transcription of FLAG fusion constructs. The aminoglycoside phosphotransferase II gene (neo-R) confers resistance to aminoglycosides such as G418, allowing for selection of stable transfectants.
Application
Cloning in a gene: pSF-CMV-NEO-NH2-PPT-3XFLAG plasmid vector contains a gene within the main multiple cloning site (NotI-ClaI). Any plasmid that we sell where the gene is this configuration will be located in the exact same position in relation to the start and stop codon of the gene. The only exceptions to this rule are fusions proteins where the fusion gene may be positioned at the front or end of the MCS to allow gene fusion.By positioning all of our genes in the same location it allows them to be transferred between plasmids using the same cloning method and restriction sites regardless of the plasmid being used from our product range. Inserting a new gene into this plasmid should be easily possible using a range of standard restriction enzyme sites that flank the gene currently in the vector.Multiple cloning site notes:In the multiple cloning site there are two important restriction sites called BsgI and BseRI sites. These sites both cut the DNA at the same position and cleave the stop codon of the gene in the multiple cloning site in this plasmid thereby producing a TA overhang. This overhang is compatible with any of our peptide or reporter fusion tag plasmids also cut with either of these enzymes. This allows seamless C-terminal fusions to be made with the gene in this multiple cloning site using a single cloning step from our C-terminal peptide and reporter tag product range. Normally the easiest method is to clone the C-terminal tag from our other plasmid products into this plasmid using BsgI or BseRI and the downstream ClaI restriction site.BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding sites. This allows them to cut the upstream stop codon in the gene in this plasmid regardless of the gene sequence.
Sequence
Please select the file type you require. For reference most cloning programs will import a .gb (Genbank) file and will show all of the plasmids features automatically when downloaded and imported.Genebank Vector Sequence FileFASTA Vector Sequence FileFull Plasmid Map
Analysis Note
To view the Certificate of Analysis for this product, please visit www.oxgene.com
Other Notes
Looking for more vector options to move your experiments forward faster? Consider a custom cloning vector designed and built by Oxford Genetics™. Find out more at Oxford Genetics - Sigma′s partner for cloning and expression vectors for molecular biology and synthetic biology applications.
Legal Information
Oxford Genetics is a trademark of Oxford Genetics Ltd
Oxford Genetics is a trademark of Oxford Genetics Ltd
Storage Class Code
12 - Non Combustible Liquids
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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