pSecTag2 A, B, & C Mammalian Expression Vectors

pSecTag2 and pSecTag2/Hygro are mammalian expression vectors designed for the secretion, purification, and detection of fusion proteins. Each vector hasRead more

Catalog Number	Quantity
V90020	20 μg

Catalog number V90020

Price (USD)

1,346.00

20 µg

Add to cart

Quantity:

20 μg

Price (USD)

1,346.00

20 µg

Add to cart

pSecTag2 and pSecTag2/Hygro are mammalian expression vectors designed for the secretion, purification, and detection of fusion proteins. Each vector has a large multiple cloning site in three reading frames to simplify cloning in frame with the N-terminal secretion signal. The vectors (Figure 1) offer the following features:

• Secretion signal from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Figure 2)
• Cytomegalovirus (CMV) promoter for high-level constitutive expression
• C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• C-terminal c-myc epitope for detection with an Anti-myc Antibody
• Bovine Growth Hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability
• SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen (e.g., COS-1, COS-7)

The pSecTag2 vectors carry the Zeocin™ resistance gene for cost-effective selection in mammalian cells. Zeocin™ selection can also be used in E. coli.

The pSecTag2/Hygro vectors have the Hygromycin B resistance gene for selection of stable mammalian cell lines.

For Research Use Only. Not for use in diagnostic procedures.

Specifications

Delivery TypeTransfection

For Use With (Application)Secreted Expression

Product TypeMammalian Expression Vector

Quantity20 μg

Selection Agent (Eukaryotic)Zeocin™

VectorpSec

Cloning MethodRestriction Enzyme/MCS

PromoterCMV

Protein TagHis Tag (6x), c-Myc Epitope Tag, IgK Leader Sequence

Unit Size20 µg

Contents & Storage

20 μg of each vector and a PSA expression control are supplied supercoiled and lyophilized. Store at -20°C. Vectors are guaranteed stable for 6 months when properly stored.

Frequently asked questions (FAQs)

I used the pSecTag2 vector and am seeing cellular but not secreted expression of the protein of interest. Can you offer some tips?

If the gene of interest has a start codon in the context of a perfect Kozak sequence, it may be preferentially translated over the vector’'s initiation codon, resulting in no leader sequence and no glycosylation, and hence no targeting to the endoplasmic reticulum and no secretion. This is rare, but it has been observed. If it is a problem, we recommend using PCR to delete the start codon.

Can I propagate my pSecTag2 vector in E. coli containing the Tn5 gene?

pSectag2 vectors have the Zeocin antibiotic-resistance marker for selection in E. coli, and any E. coli strain that contains the complete Tn5 transposable element (i.e., DH5alphaF'IQ, SURE, SURE2) encodes the ble (bleomycin) resistance gene that confers resistance to the Zeocin antibiotic. Hence, for the most efficient selection, we highly recommend choosing an E. coli strain that does not contain the Tn5 gene.

I see that the pSectag2 vectors carry the Zeocin antibiotic-resistance gene driven by the EM7 promoter. Can I use Zeocin antibiotic for selection when I propagate these vectors in E. coli?

Yes, you can use Zeocin antibiotic for selection in E. coli. However, keep in mind that for Zeocin antibiotic to be active, the salt concentration of the medium must remain low (<90 mM) and the pH must be 7.5. Prepare LB broth and LB agar plates using low-salt (5 g NaCl/liter) LB.

What is the difference between pSecTag2 and pSecTag2/Hygro vectors?

The only difference between these vectors is that the pSecTag2 vectors have the Zeocin antibiotic-resistance gene for stable selection, whereas the pSecTag2/Hygro vectors have the Hygromycin B resistance gene for stable selection.

I would like to get secreted expression of my protein. What are your recommendations?

You can insert an N-terminal secretion signal or leader sequence upstream of your gene and in-frame with the gene sequence to facilitate secreted expression of the protein. We actually offer the pSecTag2 (Cat. No. V90020) and pSecTag2/Hygro (Cat. No. V91020) vectors designed for this purpose. These vectors contain the N-terminal murine Ig kappa-chain secretion signal for secreted expression of the protein of interest.

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