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pBudCE4.1 Mammalian Expression Vector Invitrogen™

The pBudCE4.1 vector is designed for the independent expression of two genes from a single plasmid in mammalian cells. Using pBudCE4.1 to generate stable mammalian expression cell lines ensures that there is an equivalent copy number of each gene in the cell. This can eliminate variable expression due to differences in gene copy number. pBudCE4.1 provides expression cassettes withthe following features:

The CMV promoter for high-level transcription of genes with an optional c-myc epitope tag for rapid detection and 6xHis sequence for simple purification

• The human EF-1α promoter for high-level expression of genes with an optional V5 epitope tag for rapid detection and 6xHis sequence for simple purification
• The Sh ble (ZeoR) gene for efficient selection in both mammalian cells and E. coliwith the selection agent Zeocin™

TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells Invitrogen™

TOPO® TA Cloning® Dual Promoter kits are for fast, efficient cloning and subsequent in vitro transcription. These kits include the pCR™II-TOPO® TA vector with dual T7 and SP6 promoters. By eliminating time-consuming and tedious restriction site cloning, TOPO® cloning is the most reliable cloning method, featuring a 3-step protocol and 5-minute cloning reaction, and yielding up to 95% recombinants.

Convenient features of this TOPO® TA Cloning® Dual Promoter kit include:
• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and SP6 promoters for efficient in vitro transcription
• M13 forward and reverse primer sites for sequencing
• 16 convenient restriction sites, including EcoRI, flanking your insert for subsequent excision or subcloning
• Kanamycin and ampicillin resistance for your choice of selection in E. coli
• Easy blue/white colony screening for selection of recombinants
• Includes OneShot® TOP10 Competent cells for added convenience and highest cloning efficiencies

Kit Options: TOPO® TA Cloning® Dual Promoter Kits
TOPO® TA Cloning® Dual Promoter kits can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:
• General cloning: TOP10 cells (Cat. Nos. K4600-J10, K4600-01, K4600-40)
• High-efficiency cloning: TOP10 Electrocomp™ cells (Cat. Nos. K4660-01, K4660-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R cells (Cat. Nos. K4620-01, K4620-40)
• Fast growth: Mach1™ -T1R chemically competent E. coli (Cat. No. K4610-20)
• Repressor/induction needs: TOP10F’ cells (Cat. Nos. K4650-01, K4650-40)
• Provide your own cells (Cat. Nos. 451641, 450641)

pLenti6.3/V5-DEST™ Gateway™ Vector Kit Invitrogen™

Invitrogen™ 's pLenti6.3⁄V5-DEST™ Gateway® Vector Kit is part of our ViraPower™ HiPerform™ Lentiviral Gateway® Expression Kit (catalog K5330-00).

The pLenti6.3⁄V5-DEST™ Gateway® Vector Kit contains the Gateway®-adapted ViraPower™ HiPerform™ lentiviral expression vector, pLenti6.3⁄V5-DEST™ for easy recombination-based cloning and high-level expression of a target gene in dividing and non-dividing mammalian cells. The pLenti6.3⁄V5-DEST™ vector is equipped with two key genetic elements, making it a HiPerform™ vector: the Woodchuck Posttranscriptional Regulatory Element (WPRE) and the central Polypurine Tract (cPPT) sequence from the HIV-1 integrase gene to produce at least 4-fold increase in protein expression compared to vectors lacking these elements.

Advantages
• Stable expression
• Long-term experiments
• Accurate titer of functional virus
• Flexible and versatile Gateway® recombination cloning technology

Key Features
• HiPerform™ WPRE and cPPT elements
• CMV promoter
• V5 epitope tag at C terminus
• Blasticidin selection

Kit includes
• A lacZ vector as a positive control, pLenti6.3⁄V5-GW⁄lacZ
• Stbl3™ competent cells

For research use only. Not intended for any therapeutic or diagnostic use.

TOPO™ TA, Dual TOP10 (Supply Center Packaging) Invitrogen™

The TOPO® TA Cloning® Dual Promoter Kit is for fast, efficient cloning and subsequent in vitro transcription/translation. This kit includes the pCR™II-TOPO® TA vector (see figure). By eliminating time-consuming and tedious restriction site cloning, TOPO® cloning is the most reliable method, taking only 5 minutes using a 3-step protocol, and yielding up to 95% recombinants. Convenient features of the TOPO® TA Cloning® Dual Promoter Kit include:

• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and SP6 promoters for efficient in vitro transcription
• M13 forward and reverse primer sites for sequencing
• 16 convenient restriction sites, including EcoRI, flanking your insert for subsequent excision or subcloning
• Kanamycin and ampicillin resistance for your choice of selection in E. coli
• Easy blue/white colony screening for selection of recombinants
• Includes OneShot® TOP10 Competent cells for added convenience and highest cloning efficiencies

This product comes in Supply Center packaging, which features an additional outer box that holds both the TOPO® vector box and the competent cells box. This special packaging helps ensure that visitors to a Supply Center obtain the complete kit.

pCMV-Cypridina Luc Vector for Luciferase Assays Thermo Scientific™

The Thermo Scientific pCMV-Cypridina Luc Vector is a multiple cloning site plasmid designed to accept a promoter sequence for study of gene regulation using the naturally secreting Cypridina luciferase reporter.

The Cypridina Luc Vectors contain a gene cloned from the marine ostracod, Cypridina noctiluca. The gene encodes a naturally-secreted bioluminescent Cypridina luciferase (62kDa), which enables measurement of the reporter activity in media (for real-time assays) and in cell lysates. The pCMV vector has the luciferase gene under the CMV (Cytomegalovirus) promoter, and this constitutive expression vector can be used as a normalization control to account for experimental variation in combination with other reporters.

Features of the Cypridina Luc Vectors:

• Naturally-secreting Cypridina luciferase gene, optimized for high expression in mammalian systems
• Multiple cloning site (MCS) provides versatility for transfer of regulatory elements from one plasmid to another
• Transcription termination site (Ter), Lac operator (Lac O1), and transcriptional pause site (TPS) used to minimize background by reducing transcriptional read-through
• Both puromycin (Pur) and ampicillin (Amp) markers for drug selection in mammalian and bacterial cells, respectively
• High-copy pUC bacterial DNA replication origin
• Two control vectors available with strong (CMV) or weak (TK) constitutive promoters for co-transfection and normalization

The pMCS-, pCMV- and pTK-Cypridina Luc Vectors encode the Cypridina luciferase reporter with excellent light intensity.

These vectors are subject to a limited use label license.

More Product Data
Highly sensitive multiplex luciferase reporter assays
Luciferase assays in hard-to-transfect Jurkat cells
Monitoring neuronal differentiation using multiplexed luciferase reporters
Activation of the antioxidant response pathway by pesticide chemicals

Related Products
pMCS-Cypridina Luc Vector for Luciferase Assays
pTK-Cypridina Luc Vector for Luciferase Assays

pLenti7.3/V5-DEST™ Gateway™ Vector Kit Invitrogen™

The pLenti7.3⁄V5-DEST™ Gateway® Vector Kit contains the Gateway®-adapted ViraPower™ HiPerform™ lentiviral expression vector, pLenti7.3⁄V5-DEST™ for easy recombination-based cloning and high-level expression of a target gene in dividing and non-dividing mammalian cells. The pLenti7.3⁄V5-DEST™ vector is equipped with two key genetic elements, making it a HiPerform™ vector: the Woodchuck Posttranscriptional Regulatory Element (WPRE) and the central Polypurine Tract (cPPT) sequence from the HIV-1 integrase gene to produce at least 4-fold increase in protein expression compared to vectors lacking these elements. In addition, the vector allows for an accurate determination of titer of functional lentivirus in just two days using Emerald Green Fluorescent Protein (EmGFP).

Advantages
• Transient expression
• Short-term experiments
• High-throughput screening
• 2 day titer of functional virus using EmGFP
• Flexible and versatile Gateway® recombination cloning technology

Key Features
• WPRE (Woodchuck Posttranscriptional Regulatory Element) from the woodchuck hepatitis virus, allows increased transgene expression and cPPT (Polypurine Tract) from the HIV-1 integrase gene, increases the copy number of lentivirus integrating into the host genome
• Human cytomegalovirus (CMV) immediate early promoter to control high-level expression of the gene of interest
• SV40 promoter driving expression of EmGFP
• Emerald Green Fluorescent Protein (EmGFP) allows easy determination of Lentiviral titers by flow cytometry and monitor transduced cells.

Kit Includes
• pLenti7.3⁄V5-DEST™ Gateway® Vector box
• One Shot® Stbl3™ chemically competent E. coli (C7373-03)

Related SKUs
• 293FT Cell Line (R70007; R70007)
• ViraPower™ Lentiviral Support Kit (K4970-00)
• ViraPower™ Lentiviral Packaging Mix (K4975-00)
• Lipofectamine® 2000 (11668-019; 11668-027)

For research use only. Not intended for any therapeutic or diagnostic use.

pREP4 Mammalian Expression Vector Invitrogen™

These vectors are designed for high-level, constitutive expression from either the CMV or RSV promoters. Both vectors contain the EBNA-1 gene for episomal expression in primate and canine cell lines.

ViraPower™ HiPerform™ T-REx™ Gateway™ Expression System Invitrogen™

The ViraPower™ HiPerform™ T-REx™ Gateway® Expression System includes all the components needed to generate lentivirus, including vector kit, 293FT cell line, and the support kit. This kit combines Invitrogen’s ViraPower™ HiPerform™ Lentiviral, T-REx™ and Gateway® technologies to facilitate easy recombination-based cloning and lentiviral-based, regulated (Tetracycline-inducible), high-level expression of a target gene in dividing and non-dividing mammalian cells. The pLenti6.3⁄ TO⁄ V5-DEST vector is equipped with two key genetic elements, making it a HiPerform™ vector: the Woodchuck Posttranscriptional Regulatory Element (WPRE) and the central Polypurine Tract (cPPT) sequence from the HIV-1 integrase gene to produce at least 4-fold increase in protein expression compared to vectors lacking these elements.

Advantages
• Generates replication-incompetent lentivirus for transducing dividing and non-dividing mammalian cells
• Easy recombination-based cloning using Gateway® technology
• Stable, long-term, tetracycline-regulated expression
• Enhanced protein expression, up to 4-fold or greater, compared to traditional lentiviral expression systems

Key Features
• WPRE (Woodchuck Posttranscriptional Regulatory Element) from the woodchuck hepatitis virus, increases transgene expression and cPPT (central Polypurine Tract) from the HIV-1 integrase gene, increases the copy number of lentivirus integrating into the host genome, thus increasing viral titer. WPRE and cPPT together produce at least a four-fold increase in protein expression in most cell types, compared to other vectors that do not contain these elements.
• Hybrid promoter consisting of the human cytomegalovirus (CMV) promoter and two tetracycline operator 2 (TetO2) sites for high-level, regulated expression of the target gene
• Blasticidin selection marker for stable selection under control of SV40 promoter

Kit Includes
• ViraPower™ HiPerform™ T-Rex™ Gateway® Vector Kit (Cat # A11144)
• ViraPower™ Bsd Lentiviral Support Kit (Cat # K497000)
• 293FT Cell Line (Cat # R70007)
• Gateway® LR Clonase® II Plus Enzyme Mix (Cat # 12538120)
• One Shot® Stbl3™ Chemically Competent E. coli (Cat # C737303)
• Geneticin® (Cat # 10131035)
• pENTR™ Gus positive control plasmid

For research use only. Not intended for any therapeutic or diagnostic use.

pCRE Tluc16-DD Vector for Luciferase Assays Thermo Scientific™

The pCRE Tluc16-DD Vector is a transcriptional reporter vector designed to monitor the activation of cAMP-binding protein (CREB) and cAMP-mediated signal transduction pathways in mammalian cells. The vector encodes the smallest known luciferase, TurboLuc™16 (Tluc16) luciferase, as the reporter under the control of a combination of an optimized minimal core promoter and 5 tandem repeats of the c-AMP response element (CRE).

• Intracellular Tluc16 luciferase gene, optimized for high expression in mammalian systems
• Dual-destabilization (DD) technology reduces accumulation of Tluc16 luciferase mRNA and protein in cells, enhancing the responsiveness of the assay
• Monitor the activation of cAMP-binding protein (CREB) and cAMP-mediated signal transduction pathways in mammalian cells

Tluc16 luciferase is a 16 kDa, novel, intracellular luciferase derived from the marine copedod Metridia luciferase family. The wild-type luciferase has been modified to reduce its size, increase its brightness, and enable its intracellular expression.

The pCRE Tluc16-DD luciferase expression vector also contains dual-destabilization (DD) technology that reduces accumulation of the Tluc16 luciferase mRNA and protein in cells, enhancing the responsiveness of the assay. A synthetic polyA terminator and a Transcriptional Pause Site (TPS) are included upstream of the cAMP response elements to minimize non-specific transcriptional read-through. The Tluc16 luciferase activity can be measured using the Thermo Scientific TurboLuc™ One-Step Glow Assay Kit.

Vivid Colors™ pLenti6.3/V5-GW/EmGFP Expression Control Vector Invitrogen™

The Vivid Colors™ pLenti6.3⁄V5-GW⁄EmGFP Expression Control Vector is a ViraPower™ HiPerform™ positive control lentiviral vector containing Emerald Green Fluorescent Protein (EmGFP). It is designed for use with the ViraPower™ HiPerform™ Lentiviral Expression Systems as a positive control to enable the detection of higher levels of EmGFP fluorescence following transfection in 293FT cells, as a titer control to produce an EmGFP-expressing lentivirus stock and as a transduction control following transduction in both dividing and non-dividing mammalian cells. The vector has the CMV promoter for driving constitutive expression of EmGFP and the SV40 promoter for driving expression of the blasticidin stable selection marker. The vector is equipped with two key genetic elements, making them Hiperform™ vectors: the Woodchuck Posttranscriptional Regulatory Element (WPRE) and the central Polypurine Tract (cPPT) sequence from the HIV-1 integrase gene to produce at least 4-fold increase in EmGFP expression compared to vectors lacking these elements. This control vector is not designed for generating EmGFP fusion proteins and does not express the V5 epitope.

Advantages
• Lentivirus based expression of EmGFP in dividing and non-dividing mammalian cells
• Serve as a quick positive control for transfection and lentiviral production
• Serve as a quick titer control in determination of lentivirus titer

Key Features
• Constitutive expression with CMV promoter
• High level expression of EmGFP without V5 epitope
• WPRE and cPPT sequences produce at least 4-fold increase in EmGFP expression compared to other Lenti vectors without these elements
• Blasticidin selection marker for stable selection

Kit Includes
• pLenti6.3⁄V5-GW⁄EmGFP Expression Control Vector

Related SKUs
• ViraPower™ HiPerform™ Lentiviral TOPO® Expression Kit (K531000)
• ViraPower™ HiPerform™ Lentiviral Gateway® Expression Kit (K533000)
• ViraPower™ HiPerform™ T-Rex™ Gateway® Expression System (A11141)
• ViraPower™ HiPerform™ Promoterless Gateway® Expression System (A11145)

For research use only. Not intended for any therapeutic or diagnostic use.

pcDNA™3.1 Directional TOPO™ Expression Kit Invitrogen™

The pcDNA™3.1 Directional TOPO® Expression Kit uses linearized, topoisomerase I-activated pcDNA3.1D/V5-His-TOPO® for five-minute directional cloning and subsequent high-level expression.

Directional Cloning technology facilitates expression experiments because:

• A proofreading enzyme is used for fewer errors in cloned genes
• Greater than 90% of the clones are in the correct orientation for expression, reducing time spent colony screening for clone orientation

In addition, pcDNA3.1D/V5-His-TOPO® provides strong expression levels from the CMV promoter and the option of a C-terminal V5-His fusion tag for easy detection of recombinant protein with an Anti-V5 Antibody and rapid purification on nickel-chelating resin.

GeneArt™ pYES1L Vector with Sapphire™ Technology Invitrogen™

GeneArt® pYES1L Vector with Sapphire Technology™ is a ready-to-use, 9.3 Kb linearized BAC⁄YAC shuttle vector, used for the assembly of DNA fragments with the GeneArt® High-Order Genetic Assembly System (cat#A13285 & A13286). The vector has the capacity to clone up to 110 Kb of DNA fragments. This product only includes the linearized GeneArt® pYES1L Vector with Sapphire Technology™.

Some of the key features and elements of the GeneArt® pYES1L Vector with Sapphire Technology™ include:
TRP1 to allow selection of yeast transformants in tryptophan-deficient medium
• ARS⁄CEN origin to allow stable maintenance of the construct in yeast
• F’ for conjugation
• oriT for maintenance of large constructs in E.coli
• Spectinomycin resistance for selection in E.coli
• 6 Convenient and validated restriction sites flanking the cloning region

TOPO™ XL-2 Complete PCR Cloning Kit, with One Shot™ OmniMAX™ 2 T1R Chemically Competent E. coli Cells Invitrogen™

The TOPO™ XL-2 Complete PCR Cloning Kit provides all the necessary elements for efficient cloning of extra-long PCR products (up to 13 kb). The kit uses the linearized and topoisomerase 1-activated pCR™-XL-2-TOPO™ vector, which is compatible with the cloning of blunt-end PCR fragments. Amplification of long PCR fragments is enabled by Platinum™ SuperFi™ Green PCR Master Mix, which is included in the kit. Topoisomerase I activation of the vector allows PCR products to be ligated in just 5 minutes on your bench–top, resulting in high cloning efficiency (up to 90% positive recombinants with 10 kb fragments).

The pCR-XL-2-TOPO vector includes:
• ccdB gene for positive selection
• EcoR I site flanking the PCR product insertion site for easy excision of inserts
• Ampicillin- and kanamycin-resistance genes for your choice of selection in E. coli
• T7 promoter/priming site for in vitro RNA transcription and sequencing
• T7, T3, and M13 forward and reverse primer sites for sequencing

The TOPO XL-2 Complete PCR Cloning Kit includes:
• TOPO XL-2 PCR Cloning Kits containing the pCR-XL-2-TOPO vector
Platinum SuperFi Green PCR Master Mix—featuring a proofreading DNA polymerase (>300X fidelity compared to Taq) with high processivity and a density gradient to generate accurate, long PCR amplicons ready to load onto an agarose gel for gel extraction
PureLink™ Quick Gel Extraction and PCR Purification Combo Kit—designed to purify DNA fragments from agarose gels or direct PCR purification using a silica-based spin cartridge in < 30 minutes
One Shot™ OmniMAX™ 2 T1R Chemically Competent E. coli Cells—an improved chemically competent cell line, perfect for use in all cloning applications.

pOG44 Flp-Recombinase Expression Vector Invitrogen™

In addition to the original pcDNA™5/FRT vector, four more Flp-In™ expression vectors-pcDNA5/FRT/V5-His-TOPO®, pSecTag/FRT/V5-His-TOPO®, pEF5/FRT/V5-DEST™, and pEF5/FRT/V5-D-TOPO® (Figure 1)-are available for cloning and expressing in the Flp-In™ System. These vectors offer a variety of cloning options and different promoter types. Vectors include the following features:

• Flp Recombinase Target (FRT) site for efficient integration into Flp-In™ Cell Lines
• Hygromycin resistance gene for convenient selection of integrants
• C-terminal V5 tag for easy detection of fusion proteins with the Anti-V5 Antibody
• 6xHis tag (pcDNA5/FRT/V5-His-TOPO® and pSecTag/FRT/V5-His-TOPO® vectors only) for rapid purification of fusion proteins on nickel-chelating resin

All vectors feature either the high-level CMV or EF-1α promoter. In addition pSecTag/FRT/V5-His-TOPO® contains the Igκ leader sequence for secretion of recombinant protein.

Entry Options
pcDNA™5/FRT is suitable for restriction digest-mediated cloning. The other four Flp-In™ expression vectors offer two time-saving options for cloning into a Flp-In™ expression vector:
Five-minute TOPO® Cloning

The pcDNA™5/FRT/V5-His and pSecTag/FRT/V5-His TOPO® TA Expression Kits offer five-minute cloning of Taq-amplified PCR products directly into a Flp-In™ expression vector. The pEF5/FRT/V5 Directional TOPO® Expression Kit allows five-minute directional cloning of PCR product generated with a proofreading enzyme. Using Directional TOPO® Cloning, >90% of the resulting clones will be in the correct orientation for expression, reducing time spent colony screening.
Easy Recombination into a Gateway® vector

The pEF5/FRT/V5-DEST™ vector is compatible with Gateway® Technology* for easy transfer of your gene of interest into different vectors or host systems. The fast, efficient Gateway® recombination reaction can be simultaneously performed with multiple destination vectors, saving you time when working with different systems.

Choice of Promoters
Flp-In™ expression vectors are available with the CMV or EF-1α promoter. The pEF5/FRT/V5-DEST™ and pEF5/FRT/V5-D-TOPO® vectors contain the human EF-1É promoter for driving expression of the gene of interest. The EF-1α promoter expresses in a wide range of mammalian cell types, including those where the CMV promoter expression is absent or inconsistent. This promoter may be more appropriate for long-term gene expression in some cell types and is recommended for expression in the BHK and 3T3 pre-made Flp-In™ Cell Lines. The pcDNA5/FRT/V5-His-TOPO® and pSecTag/FRT/V5-His-TOPO® vectors carry the CMV promoter for high-level constitutive expression of your gene of interest in most cell types.

pCEP4 Mammalian Expression Vector Invitrogen™

These vectors are designed for high-level, constitutive expression from either the CMV or RSV promoters. Both vectors contain the EBNA-1 gene for episomal expression in primate and canine cell lines.
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