OGS626
pSF-CMV-NEO-NH2-PPT-3XFLAG
(plasmid vector for molecular cloning)
Synonyme(s) :
cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, snapfast vector, vector
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About This Item
Code UNSPSC :
12352200
Nomenclature NACRES :
NA.85
Produits recommandés
Produit recombinant
expressed in human cells
Étiquette/Marqueur
3X FLAG tagged
Forme
buffered aqueous solution
Poids mol.
size 5955 bp
Sélection de bactéries
ampicillin
Origine de la réplication
pUC
Clivage des peptides
EKT
Localisation du marqueur peptidique
N-terminal
Promoteur
Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian
Conditions d'expédition
ambient
Température de stockage
−20°C
Description générale
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.
Séquence
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
Remarque sur l'analyse
To view the Certificate of Analysis for this product, please visit www.oxgene.com
Autres remarques
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.
Informations légales
Oxford Genetics is a trademark of Oxford Genetics Ltd
Oxford Genetics is a trademark of Oxford Genetics Ltd
Code de la classe de stockage
12 - Non Combustible Liquids
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
Certificats d'analyse (COA)
Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".
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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.
Diana Romero et al.
Carcinogenesis, 37(1), 18-29 (2015-10-28)
Dickkopf-3 (Dkk-3) is a secreted protein whose expression is downregulated in many types of cancer. Endogenous Dkk-3 is required for formation of acini in 3D cultures of prostate epithelial cells, where it inhibits transforming growth factor (TGF)-β/Smad signaling. Here, we
Geoffrey M Lynn et al.
Nature biotechnology, 33(11), 1201-1210 (2015-10-27)
The efficacy of vaccine adjuvants such as Toll-like receptor agonists (TLRa) can be improved through formulation and delivery approaches. Here, we attached small molecule TLR-7/8a to polymer scaffolds (polymer-TLR-7/8a) and evaluated how different physicochemical properties of the TLR-7/8a and polymer
Jin-Gyoung Jung et al.
PLoS genetics, 10(10), e1004751-e1004751 (2014-10-31)
The Notch3 signaling pathway is thought to play a critical role in cancer development, as evidenced by the Notch3 amplification and rearrangement observed in human cancers. However, the molecular mechanism by which Notch3 signaling contributes to tumorigenesis is largely unknown.
Alexander C Cerny et al.
PLoS genetics, 11(10), e1005578-e1005578 (2015-10-29)
Recycling of signaling proteins is a common phenomenon in diverse signaling pathways. In photoreceptors of Drosophila, light absorption by rhodopsin triggers a phospholipase Cβ-mediated opening of the ion channels transient receptor potential (TRP) and TRP-like (TRPL) and generates the visual
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