Promega's Cookie Policy

We use cookies and similar technologies to make our website work, run analytics, improve our website, and show you personalized content and advertising. Some of these cookies are essential for our website to work. For others, we won’t set them unless you accept them. To find out more about cookies and how to manage cookies, read our Cookie Policy.

CloneWeaver® Workflow Purchasing Tool

Powering Your Cloning Workflow

Product selection has never been easier

Molecular Cloning is the process of producing recombinant DNA and transforming into host organisms to replicate and make more copies. Every cloning project is unique. Read More

Once you have designed the cloning scheme, gathering all of the required reagents to get you from construct to expression and analysis is not trivial. CloneWeaver helps you build a customized cloning "kit" with all of the items your cloning scheme requires. Purchase your selection instantly, save it or email it to a purchasing agent. Soon you'll have everything you require to create the construct you need.

Previous Next

Vectors: Mammalian Expression Change

Name Description Part Number
CheckMate Mammalian Two-Hybrid System Open/Close Add
Mammalian system for studying interactions in cell line of choice
Study post-translational modifications including glycosylation, phosphorylation and acylation

Two-hybrid systems are extremely powerful methods for detecting protein:protein interactions in vivo. The basis of two-hybrid systems is the modular domains found in some transcription factors: a DNA-binding domain, which binds to a specific DNA sequence, and a transcriptional activation domain, which interacts with the basal transcriptional machinery. A transcriptional activation domain in association with a DNA-binding domain will promote the assembly of RNA polymerase II complexes at the TATA box and increase transcription. In the CheckMate Mammalian Two-Hybrid System the DNA-binding domain and the transcriptional activation domain, produced by separate plasmids, are closely associated when one protein (X) fused to a DNA-binding domain interacts with a second protein (Y) fused to a transcriptional activation domain. In this system, interaction between proteins X and Y results in transcription of a reporter gene.

Visit Product Page »
CheckMate/Flexi Vector Mammalian Two-Hybrid System Open/Close Add
Confirm, validate and study suspected protein or domain interactions
Generate stable cell lines for cell-based assays
Enables mammalian protein expression and post-translational modification studies in situ

The CheckMate/Flexi Vector Mammalian Two-Hybrid System provides a means to confirm, validate and study suspected interactions between two proteins or domains and can be used to generate stable cell lines for cell-based assays. Developed primarily for mammalian proteins of interest, the system can allow protein expression and post-translational modifications in an environment mimicking the native cell milieu. It is patterned on the yeast two-hybrid system with one protein of interest (X) fused to a DNA-binding domain and the other protein (Y) fused to a transcriptional activation domain. The system relies upon three plasmids that are co-transfected into mammalian cells, each plasmid having unique features. This system differs from the original CheckMate Mammalian Two-Hybrid System in that the vectors are compatible with the Flexi Vector System, which allows directional cloning and rapid, efficient and high-fidelity transfer of protein coding regions between a variety of Flexi Vectors.

Visit Product Page »
pAdVAntage Vector Open/Close Add
Increases translation initiation in cells transiently transfected with protein expression constructs
Can be used in a variety of cell lines

Co-transfection of mammalian cells with the pAdVAntage Vector enhances transient protein expression in a variety of cell types by increasing translation initiation. Transfection of mammalian cells with an expression vector often results in suboptimal expression of the protein of interest. Double-stranded RNA (dsRNA) generated during transfection is thought to activate the dsRNA-activated inhibitor (DAI), one of several enzymes involved in the host cell's antiviral defense system. DAI phosphorylates the translation initiation factor eIF-2, halting translation and therefore protein production. However, DAI translation inhibition can be overcome with the adenoviral Virus Associated I RNA (VAI RNA) produced by RNA polymerase III following co-transfection with the pAdVAntage Vector. The VAI RNA binds to DAI, preventing its activation, thereby allowing translation and protein expression.

Visit Product Page »
pCI-neo Mammalian Expression Vector Open/Close Add
Select for stably transfected cells with neomycin phosphotransferase gene
Increase the steady-state level of RNA with the late SV40 polyadenylation signal approximately fivefold more than the early SV40 polyadenylation signal
Synthesize transcripts in vitro using the T7 RNA polymerase promoter or generate ssDNA in E. coli using the f1 origin of replication

The pCI-neo Mammalian Expression Vector carries the human cytomegalovirus (CMV) immediate-early enhancer/promoter region to promote constitutive expression of cloned DNA inserts in mammalian cells. This vector also contains the neomycin phosphotransferase gene, a selectable marker for mammalian cells. The pCI-neo Vector can be used for transient or stable expression by selecting transfected cells with the antibiotic G-418.

Visit Product Page »
pCMVTNT and pTNT Vectors Open/Close Add
Express cloned genes in vitro or in vivo from SP6- or T7-based coupled in vitro transcription/translation systems

The pCMVTNT and pTNT Vectors are designed for convenient expression of cloned genes in vitro or in vivo. SP6 and T7 promoters allow expression from SP6- or T7-based coupled in vitro transcription/translation systems. The presence of RNA phage promoters also allows highly efficient synthesis of RNA in vitro. Both vectors contain a 5´ beta-globin leader sequence and synthetic poly(A)30 tail, which have been shown to enhance expression of certain genes. For in vivo expression, the pCMVTNT Vector contains a CMV enhancer/promoter region, which allows strong constitutive expression in many cell types.

Visit Product Page »
pFN21A and pFN21K HaloTag CMV Flexi Vectors Open/Close Add
Enables constitutive protein expression in mammalian cells
Includes CMV immediate-early enhancer/promoter
Suited to stable or transient gene expression

These vectors are designed for expression of N-terminal-tagged HaloTag fusion proteins in mammalian cells. Once expressed, the HaloTag fusion protein may be used for cell imaging of protein localization or trafficking in conjunction with the fluorescent HaloTag Ligands. In addition, the HaloTag fusion protein can be purified or pulled down as a complex with its protein partners. We offer two types of HaloTag fusion vectors to accommodate your cloning preferences: pHT Vector Series: Simple Multiple Cloning Site (MCS) plasmids for traditional cloning. pF Vector Series: Flexi Vector Cloning System---a directional cloning method for protein-coding sequences. It is based on two rare-cutting restriction enzymes, SgfI and PmeI, and provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between a variety of Flexi Vectors without the need to resequence.

Visit Product Page »
pFN28A and pFN28K HaloTag CMV-neo Flexi Vectors Open/Close Add
Enables constitutive protein expression in mammalian cells
Includes CMV immediate-early enhancer/promoter
Suited to stable or transient gene expression

These vectors are designed for expression of N-terminal-tagged HaloTag fusion proteins in mammalian cells. Once expressed, the HaloTag fusion protein may be used for cell imaging of protein localization or trafficking in conjunction with the fluorescent HaloTag Ligands. In addition, the HaloTag fusion protein can be purified or pulled down as a complex with its protein partners. We offer two types of HaloTag fusion vectors to accommodate your cloning preferences: pHT Vector Series: Simple Multiple Cloning Site (MCS) plasmids for traditional cloning. pF Vector Series: Flexi Vector Cloning System---a directional cloning method for protein-coding sequences. It is based on two rare-cutting restriction enzymes, SgfI and PmeI, and provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between a variety of Flexi Vectors without the need to resequence.

Visit Product Page »
pTARGET Mammalian Expression Vector System Open/Close Add
Clone PCR products with T overhangs
CMV promoter for robust, consistent expression
Neomycin marker for selection of stable transfectants

The pTARGET Mammalian Expression Vector System is a convenient system for cloning PCR products and expressing cloned PCR products in mammalian cells. Prepare the vector by digesting with EcoRV and adding a 3' terminal thymidine to each end. These single 3'-T overhangs at the insertion site greatly improve the efficiency of ligation of a PCR product into the plasmid in two ways. First, the overhangs prevent recircularization of the vector; second, they provide a compatible overhang for PCR products generated by thermostable polymerases that add a single deoxyadenosine, in a template-independent fashion, to the 3'-ends of amplified fragments. This vector contains a modified coding sequence of the alpha-peptide of beta-galactosidase, which allows recombinants to be selected using blue/white screening, and carries the human cytomegalovirus (CMV) immediate-early enhancer/promoter region to promote constitutive expression of cloned DNA inserts in mammalian cells.

Visit Product Page »

Save

Your selections can be accessed anytime at the link you will receive via email:


Share

To share your selections via email, complete the short form below: