Sort By

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.

Bac-to-Bac™ HT Vector Kit (Gibco™)

The Bac-to-Bac® HT Vector is designed for use as part of the Bac-to-Bac® Baculovirus Expression System (Cat. No. 10359-016) for the expression and purification of histidine-tagged recombinant proteins in Sf9, Sf21, or High Five™ Cells following bacmid generation in E. coli. The pFastBac™ HT vector offers the following features:

• Strong polyhedrin promoter for protein expression
• Three reading frames for simplified cloning
• N-terminal 6xHis tag for simple purification of recombinant fusion proteins
• TEV protease cleavage site for removal of the histidine tag following protein purification

pTK-Green Renilla Luc Vector for Luciferase Assays (Thermo Scientific™)

The Thermo Scientific pCMV-Green Renilla Luc vector is a derivative of pMCS-Green Renilla Luc. It contains the luciferase gene under the control of the Herpes Simplex Virus (HSV) thymidine kinase (TK) promoter, creating a constitutive expression vector that can be used as a normalization control to account for experimental variation in combination with other reporters.

Features of the pTK-Green Renilla Luc vector:

• Weak (TK) constitutive promoter for co-transfection and normalization
• Intracellular Green Renilla luciferase gene, optimized for high expression in mammalian systems
• 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

More Product Data
Luciferase assays in hard-to-transfect Jurkat cells

Related Products
pMCS-Green Renilla Luc Vector for Luciferase Assays
pCMV-Green Renilla Luc Vector for Luciferase Assays

pEXP4-DEST Vector Kit (Invitrogen™)

Expressway™ Tag-On-Demand™ Cell-Free E. coliExpression System with Lumio™ Technology enables convenient, in vitro expression of tagged or untagged protein from a single vector, real-time detection of protein synthesis, and easy in-gel protein detection. With this system, you can:

• Quickly screen for expression levels of full-length protein
• Save time and effort by cloning only once to express tagged or native protein
• Express a C-terminally tagged protein from an Ultimate™ ORF Clone in an in vitro system

The Expressway™ Tag-On-Demand™ System includes a specialized E. colilysate derived from a slyD mutant for cell-free synthesis. This lysate eliminates non-specific binding of the Lumio™ Green Detection Reagent to the endogenous SlyD protein, providing an optimal background for recombinant protein detection.

How it works
Simply clone your gene of interest containing a TAG (amber) stop codon into the provided pEXP4-DEST (Figure 1). The pEXP4-DEST vector features attR sites for efficient Gateway® recombination, C-terminal Lumio™ and 6xHis tags for simplified identification and purification, and components for cell-free protein synthesis. In the presence of the Expressway™ Tag-On-Demand™ Suppressor tRNA, the TAG codon is not recognized and translation continues through the Lumio™ and 6xHis tags. In the absence of the Suppressor tRNA, the TAG stop codon is recognized and native protein is produced (Figure 2). From the same vector, you can rapidly determine the expression level of the C-terminal-tagged, full-length protein, and then express the native protein free from any adverse affects the tag may have on its structure, activity, or molecular interactions.

pTK-Red Firefly Luc Vector for Luciferase Assays (Thermo Scientific™)

Thermo Scientific pTK-Red Firefly Luc is a constitutive expression vector having the luciferase gene under the Herpes Simplex Virus (HSV) thymidine kinase (TK) promoter for co-transfection and normalization control to account for experimental variation in combination with other reporters in a gene regulation study using the intracellular red firefly luciferase reporter with excellent light intensity..

Features of Red Firefly Luc:
• Red Firefly Luc Vectors contains a mutant form of the firefly luciferase gene that has a red-shifted emission spectrum.
• pMCS vector contains a multiple cloning site for cloning a promoter to study its regulatory potential.
• pCMV and pTK vectors have the luciferase gene under the CMV (Cytomegalovirus) promoter and HSV-TK promoter, respectively.
• These pCMV and pTK constitutive expression vectors can be used as normalization controls to account for experimental variation in combination with other reporters.

These vectors are subject to a limited use label license.

More Product Data
Highly sensitive multiplex luciferase reporter assays
Monitoring neuronal differentiation using multiplexed luciferase reporters
Activation of the antioxidant response pathway by pesticide chemicals

Related Products
pMCS-Red Firefly Luc Vector for Luciferase Assays
pCMV-Red Firefly Luc Vector for Luciferase Assays

pT7CFE1-Cftag Vector for Mammalian Cell-Free Protein Expression (Thermo Scientific™)

Thermo Scientific pT7CFE1-CFtag is a cloning plasmid optimized to use with the Thermo Scientific 1-Step Human In Vitro Protein Expression System for in vitro translation (IVT) of tagged fusion proteins. pT7CFE1-CFtag Vector is available with single affinity Ftag tag at the C-terminus to facilitate protein purification and detection.

Features of pT7CFE1:
• EMCV IRES at the 5' UTR promotes high-level translation of mRNAs
• MCS accommodates gene insertion via ten different restriction sites: Msc1, Nde1, BamH1, EcoR1, EcoRV, Pac1, Pst1, Sac1, Sal1, Not1 and Xho1
• Poly A sequence in the 3' region promotes mRNA stabilization and protection from nucleases
• T7 terminator ensures synthesis of accurate size mRNA transcripts
• Plasmid linearization may be accomplished with restriction sites between Poly A sequence and the T7 terminator region

pT7CFE1 Expression Vectors contain the Encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES) element that is critical for high levels of cap-independent protein expression in the Human In Vitro Translation System. Each vector features a highly-compatible multiple cloning site (MCS) to facilitate easy insertion of protein coding sequences into and between vectors. The pT7CFE1 Vector is available with single or tandem affinity tags at the N- or C- terminus to facilitate protein purification and detection. The pT7CFE Vectors are suitable for insertion of cloned genes, cDNAs, ORFs or PCR products for in vitro transcription and translation. Custom cloning services are also available.

More Product Data
Choosing a vector and purification method for in vitro protein expression

Related Products
pT7CFE1-NFtag Vector for Mammalian Cell-Free Protein Expression
pT7CFE1-CGST-HA-His Vector for Mammalian Cell-Free Protein Expression
pT7CFE1-NHis-GST Vector for Mammalian Cell-Free Protein Expression
pT7CFE1-NHis-GST-CHA Vector for Mammalian Cell-Free Protein Expression

pLenti4/V5-DEST™ Gateway™ Vector (Invitrogen™)

The pLenti4⁄V5-DEST™ Gateway® Vector is a Gateway®-adapted ViraPower™ lentiviral expression vector for lentiviral-based expression of a target gene in dividing and non-dividing mammalian cells. The vector has the CMV promoter for driving constitutive expression of the target gene and the Zeocin™ selection marker for stable selection in mammalian cells.

Advantages
• Lentivirus based expression of a target gene in dividing and non-dividing mammalian cells

Key Features
• Flexible and versatile Gateway® recombination cloning technology
• Constitutive high expression with CMV promoter
• Zeocin™ selection marker for stable selection
• C terminal V5 tag for quick detection

Kit includes
• pLenti4⁄V5-DEST™ Gateway® Vector
• One Shot® Stbl3™ Chemically Competent E. coli (C7373-03)

Related SKUs
• pLenti6⁄UbC⁄V5-DEST™ Gateway® Vector (V49910)
• pLenti6⁄V5-DEST™ Gateway® Vector (V49610)
• pLenti6⁄V5 Directional TOPO® Cloning Kit (K4955-10)
• ViraPower™ Lentiviral Directional TOPO® Expression Kit (K495000)
• ViraPower™ Lentiviral Gateway® Expression Kit (K4960-00)
• ViraPower™ HiPerform™ Lentiviral Gateway® Expression Kit (K5330-00)

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

pIZ/V5-His Vector Kit (Invitrogen™)

The InsectSelect™ Kit features the pIZ/V5-His vector for high-level expression in a variety of insect cells. This exceptionally small vector has several features that facilitate expression, analysis, and detection of recombinant protein in insect cells:

• The OpIE2 promoter for constitutive expression
• The Zeocin™ resistance gene for rapid selection of stably transfected cell lines
• C-terminal V5 epitope and polyhistidine (6xHis) sequence for detection with Invitrogen's Anti-V5 Antibody and rapid purification using nickel-chelating resin

BacMam pCMV-Dest Vector (Invitrogen™)

The BacMam pCMV-Dest Vector combines Gateway® cloning and BacMam gene expression technologies for easy recombination-based cloning and baculovirus-based expression of a target gene in variety of cell types. With BacMam technology, a modified insect cell virus (baculovirus) is used as a vehicle to efficiently deliver and transiently express genes in mammalian cells with minimum effort and toxicity. This technology allows for control over the level of expression through the increase or decrease of viral particle concentration. When combined with Gateway® cloning technology, a variety of sizes of target genes can be cloned and expressed. Sizes ranging from an insert for RNAi to a large gene with introns (38 kb) have been cloned and expressed successfully using the BacMam pCMV-Dest Vector. This plasmid accommodates various cloning schemes, including Gateway® , Seamless Assembly, and traditional cloning using restriction enzymes.

The BacMam pCMV-Dest Vector contains a CMV promoter to drive constitutive expression of the target gene. It also contains VSV-G elements, which enable baculovirus to have high transduction efficiency, and a WPRE element, which elongates transient expression in mammalian systems. To facilitate successful cloning, AmpR and gentamicin have been incorporated for positive selection, as well as Cm(R) (chloramphenicol resistance gene) for negative selection. Upon successful cloning, the region containing the CmR is replaced by the insert, selecting against any bacteria that still have a CmR-containing region.

TOPO™ TA Cloning™ Kit, with pCR™2.1-TOPO™, One Shot TOP10 Chemically Competent E. coli, and PureLink™ Quick Plasmid Miniprep Kit (Invitrogen™)

TOPO® TA Cloning® Kits are designed for cloning PCR products directly from a PCR reaction in just 5 minutes (1). They use a pCR™-TOPO® Vector with covalently bound topoisomerase I for fast cloning and recombinants. pCR™-TOPO® Vectors include:
• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• Choice of T7 (pCR™2.1-TOPO®) or T7 and SP6 (pCR™II-TOPO®) promoters for in vitro RNA transcription and sequencing. Each vector also contains M13 forward and reverse primer sites for sequencing.
EcoR I sites flanking the PCR product insertion site for easy excision of inserts
• Kanamycin and ampicillin resistance genes for your choice of selection in E. coli
• Easy blue/white colony screening for selection of recombinants

TOPO® TA Cloning® Kits are available in combo kits combined with a PureLink™ Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO®-cloned inserts for downstream analysis.

pcDNA™6.2/V5-PL-DEST Mammalian Expression Vector (Invitrogen™)

The pcDNA™ vectors are designed for high-level, constitutive expression in a variety of mammalian cell lines. The pcDNA6.2/V5-pL-DEST vector offers the following key features:

•Promoterless version of the pcDNA™6.2⁄V5-DEST vector (cat. no. 12489027)
attR sites for Gateway® cloning
•Compatible with MultiSite Gateway® Pro kits (e.g. cat. no. 12537100)
•C-terminal V5 tag for easy detection
•Blasticidin resistance gene for efficient stable selection
•Ampicillin resistance gene and pUC origin for selection and maintenance in E. coli

Gateway® Cloning
To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ ORF Clone. The following table lists a variety of available destination vectors.

Additional materials required, available separately: Gateway® entry clone, appropriate Gateway® LR Clonase® enzyme mix, and reaction buffer.

pMCS-Green Renilla Luc Vector for Luciferase Assays (Thermo Scientific™)

The Thermo Scientific pMCS-Renilla Luc vector is a multiple cloning site plasmid to designed to accept a promoter sequence for study of gene regulation using the intracellular Renilla luciferase reporter.

Features of the pMCS-Renilla Luc vector:

• Intracellular Green Renilla 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

The Green Renilla Luc Vectors contain a gene cloned from the sea pansy, Renilla reniformis. The gene encodes a bioluminescent Green Renilla luciferase (36kDa). The pMCS vector contains a multiple cloning site for cloning a promoter to study its regulatory potential.

These vectors are subject to a limited use label license.

More Product Data
Luciferase assays in hard-to-transfect Jurkat cells

Related Products
pCMV-Green Renilla Luc Vector for Luciferase Assays
pTK-Green Renilla Luc Vector for Luciferase Assays

pMCS-Red Firefly Luc Vector for Luciferase Assays (Thermo Scientific™)

Thermo Scientific pMCS-Red Firefly Luc is a multiple cloning site plasmid designed to accept a promoter sequence for the study of gene regulation using the intracellular red firefly luciferase reporter with excellent light intensity..

Features of pMCS-Red Firefly Luc:

• Intracellular Red Firefly 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

Features of Red Firefly Luc:
• Red Firefly Luc Vectors contains a mutant form of the firefly luciferase gene that has a red-shifted emission spectrum.
• The pMCS vector contains a multiple cloning site for cloning a promoter to study its regulatory potential.
• The pCMV and pTK vectors have the luciferase gene under the CMV (Cytomegalovirus) promoter and Herpes Simplex Virus (HSV) thymidine kinase (TK) promoter, respectively.
• These pCMV and pTK constitutive expression vectors can be used as normalization controls to account for experimental variation in combination with other reporters.

These vectors are subject to a limited use label license.

More Product Data
Highly sensitive multiplex luciferase reporter assays
Monitoring neuronal differentiation using multiplexed luciferase reporters
Activation of the antioxidant response pathway by pesticide chemicals

Related Products
pCMV-Red Firefly Luc Vector for Luciferase Assays
pTK-Red Firefly Luc Vector for Luciferase Assays

pAd/BLOCK-iT™-DEST RNAi Gateway Vector (Invitrogen™)

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately: Gateway® entry clone, Gateway® LR Clonase® enzyme mix, and reaction buffer.

TA Cloning™ Kit, with pCR™2.1 Vector and One Shot™ INVαF' Chemically Competent E. coli (Invitrogen™)

The TA Cloning® Kit with pCR™2.1 vector provides a quick, one-step cloning strategy for directly inserting a Taq-amplified PCR product into a plasmid vector. The TA Cloning® Kit uses the pCR™2.1 cloning vector and ExpressLink™ T4 DNA Ligase to generate a ligation product in a fifteen-minute, room-temperature ligation step. Reactions typically yield >80% recombinants containing inserts.

Features of the TA Cloning® Kit with pCR™2.1 vector:
Fast & convenient—15-minute, room-temperature ligation
Efficient—blue/white screening and >80% clones with correct insert
Flexible—choice of kanamycin or ampicillin resistance for flexible antibiotic selection
Hassle-free—eliminates any enzymatic modifications of the PCR product
Streamlined—does not require the use of PCR primers that contain restriction sites

The pCR™2.1 vector provides:
• 3'-T overhangs for direct ligation of Taq-amplified PCR products
• T7 promoter for in vitro RNA transcription and sequencing
• A versatile polylinker with flanking EcoR I sites for easy excision of inserts
• M13 forward and reverse primer sites for sequencing

How TA Cloning® Works
Taq polymerase has a non-template-dependent activity that adds a single deoxyadenosine (A) to the 3' ends of PCR products. The linearized vector supplied in this kit has single 3' deoxythymidine (T) residues. This allows PCR inserts to ligate efficiently with the vector.

Kit Configurations
The TA Cloning® Kit is offered in a variety of configurations: without competent cells (K2020-20 and K2020-40), with One Shot® INVF' Chemically Competent E. coli (K2000-01 and K2000-40), with One Shot® TOP10F' Chemically Competent E. coli (K2030-01 and K2030-40), and with One Shot® TOP10 Chemically Competent E. coli (K2040-01 and K2040-40) in 20- and 40-reaction kit sizes.

Vivid Colors™ pcDNA™6.2/EmGFP-Bsd/V5-DEST Vector (Invitrogen™)

The Vivid Colors™ pcDNA™ 6.2/EmGFP-Bsd/V5-DEST vector allows you to rapidly clone your gene using Gateway® technology and simultaneously express it with Emerald Green Fluorescent Protein (EmGFP) for easy identification of transfected cells. The EmGFP is derived from well-characterized fluorescent proteins of the jellyfish Aequorea victoria (1, 2) and has been humanized for optimal mammalian expression (3) (Figure 1). If cloned with the TAG (amber) codon, your gene will be expressed in its native configuration and fully compatible with Tag-On-Demand" technology. If cloned without a stop codon, a V5-epitope tag will be fused to the carboxy-terminal end of your protein of interest. In either case, expression of EmGFP provides a bright fluorescent marker to easily identify cells co-expressing your protein. This marker allows you to focus your functional analysis on specific cells, or to sort and enrich for transfected cells in a derived population.

The Vivid Colors™ pcDNA™ 6.2/EmGFP-Bsd/V5-DEST vector provides:

• PGK promoter for high-level expression of EmGFP fused to the Blasticidin (bsd)-resistance marker
• CMV promoter for high-level expression of your protein
• V5-epitope tag option to fuse to the C-terminal end of your protein, if desired

Gateway™ pDEST™20 Vector (Invitrogen™)

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately: Gateway® entry clone, Gateway® LR Clonase® enzyme mix, and reaction buffer.

pDisplay™ Mammalian Expression Vector (Invitrogen™)

pDisplay™ is a mammalian expression vector designed to target recombinant proteins to the surface of mammalian cells. Displayed proteins can be analyzed for their ability to interact with known or putative ligands. Proteins of interest are targeted and anchored to the cell surface by cloning the gene of interest in frame with the vectors unique N-terminal secretion signal and the C-terminal transmembrane anchoring domain of platelet-derived growth factor receptor (PDGFR). In contrast to phage display vectors which operate exclusively in prokaryotic cells, the pDisplay™ vector offers the advantage of having the displayed protein of interest processed in mammalian cells. Therefore, recombinant proteins of eukaryotic origin that are expressed from pDisplay™ more closely resemble their native form. This may favor more accurate ligand binding interactions. In addition to the N-terminal cell surface targeting signal and C-terminal transmembrane anchoring domain, the pDisplay™ vector contains the following key features:

• T7 promoter/priming site for in vitro transcription of sense RNA and for sequencing of inserts
• Neomycin resistance marker for stable selection in mammalian cells using Geneticin®
• SV40 origin for replication and simple vector rescue in cell lines expressing the large T antigen (e.g., COS-1 and COS-7)
• Ampicillin resistance gene for selection in E. coli

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

pEXP1-DEST Vector Kit (Invitrogen™)

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately: Gateway® entry clone, Gateway® LR Clonase® enzyme mix, and reaction buffer.

pT7CFE1-CGST-HA-His Vector for Mammalian Cell-Free Protein Expression (Thermo Scientific™)

Thermo Scientific pT7CFE1-CGST-HA-His is a cloning plasmid optimized to use with the Thermo Scientific 1-Step Human In Vitro Protein Expression System for in vitro translation (IVT) of tagged fusion proteins. pT7CFE1-CGST-HA-His Vector is available with tandem affinity tags, GST,HA and 6xHis at the C-terminus, to facilitate protein purification and detection. pT7CFE1-CGST-HA-His also has a cleavable tag, HRV 3C, available on the C-terminus.

Features of pT7CFE1:
• EMCV IRES at the 5' UTR promotes high-level translation of mRNAs
• MCS accommodates gene insertion via ten different restriction sites: Msc1, Nde1, BamH1, EcoR1, EcoRV, Pac1, Pst1, Sac1, Sal1, Not1 and Xho1
• Poly A sequence in the 3' region promotes mRNA stabilization and protection from nucleases
• T7 terminator ensures synthesis of accurate size mRNA transcripts
• Plasmid linearization may be accomplished with restriction sites between Poly A sequence and the T7 terminator region

pT7CFE1 Expression Vectors contain the Encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES) element that is critical for high levels of cap-independent protein expression in the Human In Vitro Translation System. Each vector features a highly-compatible multiple cloning site (MCS) to facilitate easy insertion of protein coding sequences into and between vectors. The pT7CFE1 Vector is available with single or tandem affinity tags at the N- or C- terminus to facilitate protein purification and detection. The pT7CFE Vectors are suitable for insertion of cloned genes, cDNAs, ORFs or PCR products for in vitro transcription and translation. Custom cloning services are also available.

More Product Data
Choosing a vector and purification method for in vitro protein expression

Related Products
pT7CFE1-NHA-CHis Vector for Mammalian Cell-Free Protein Expression
pT7CFE1-CGFP-HA-His Vector for Mammalian Cell-Free Protein Expression
pT7CFE1-NHis-GST Vector for Mammalian Cell-Free Protein Expression
pT7CFE1-NHis-GST-CHA Vector for Mammalian Cell-Free Protein Expression

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.

pShooter™ Mammalian Expression Vector (pCMV/myc/nuc) (Invitrogen™)

The pShooter™ vectors are designed to localize recombinant proteins to the nucleus, mitochondria, endoplasmic reticulum, or cytoplasm. In addition to the localization signal, each pShooter™ vector contains:

• Strong mammalian promoter (CMV or EF-1É) for constitutive expression of your protein of interest
• C-terminal c-myc epitope for detection with an Anti-myc Antibody
• Bovine Growth Hormone (BGH) polyadenylation site for mRNA stability
• f1 origin for single-stranded rescue of sense DNA
• Neomycin resistance gene for selection of mammalian cells with Geneticin¤ selection agent
• Ampicillin resistance and pUC origin for selection and maintenance in E. coli

Each vector is provided with a positive control plasmid. The positive control expresses GFP and targets it to the same subcellular location as the pShooter™ vector.

Antibody-Expressing Positive Control Vector (Gibco™)

The Antibody-Expressing Positive Control Vector is a mammalian expression control that expresses a complete, full-length rabbit IgG. It can be used as a positive expression control for transient expression systems such as the Expi293™ Expression System, the FreeStyle™ 293 Expression System, and the FreeStyle™ MAX CHO Expression System.

Expresses rabbit IgG at ~250 mg/L in the Expi293™ Expression System
Contains sufficient material for transfection of up to 150 mL of suspension culture in the Expi293™ and FreeStyle™ systems
Protocols are provided to quantitate rabbit IgG expression from the Positive Control Vector

The Antibody-Expressing Positive Control Vector contains an optimized mix of IgG heavy chain and light chain genes. This control is included in the Expi293™ Expression System Kit.

pJTI™ R4 EXP CMV-TO EmGFP pA Vector

The pJTI™ R4 Exp CMV-TO EmGFP pA vector is a positive control vector for assessing retargeting efficiency when retargeting a Jump-In™ T-REx™ parental cell line. When co-transfected with the integrase vector (pJTI™ R4 Int vector included in the Jump-In™ parental kits) and after antibiotic selection, the EmGFP can be inducibly expressed with doxycycline and the successfully retargeted cells will fluoresce green.

Ensure the Success of Your Jump-In™ T-REx™ Retargeting Reactions
Successful retargeting of Jump-In™ T-REx™ parental cell lines like the Jump-In™ T-REx™ HEK293 Kit is dependent on a variety of factors, such as:

• Transfection efficiency
• Cell confluency
• Antibiotic selection conditions
• Quality and concentration of DNA
• Retargeting vector to integrase vector ratio

We strongly recommend including a positive control retargeting reaction using the pJTI™ R4 Exp CMV-TO EmGFP pA vector in your Jump-In™ experiment, along with negative controls (no plasmid DNA, no integrase vector), so you can easily visualize the results and optimize the retargeting conditions.

TA Cloning™ Kit, Dual Promoter, with pCR™II Vector and One Shot™ TOP10F' Chemically Competent E. coli (Invitrogen™)

The TA Cloning® Kit Dual Promoter with pCR™II vector provides a quick, one-step cloning strategy for directly inserting a Taq-amplified PCR product into a plasmid vector. The T7 and Sp6 promoters of the pCR™II vector allow in vitro transcription of the insert to produce sense or anti-sense products. The TA Cloning Kit Dual Promoter uses the pCR™II cloning vector and ExpressLink™ T4 DNA Ligase to generate a ligation product in a fifteen-minute, room-temperature ligation step. Reactions typically yield >80% recombinants containing inserts.

Features of the TA Cloning® Kit Dual Promoter:
Fast & convenient—15-minute, room-temperature ligation
Efficient—blue/white screening and >80% clones with correct insert
Flexible—choice of kanamycin or ampicillin resistance for flexible antibiotic selection
Hassle-free—eliminates any enzymatic modifications of the PCR product
Streamlined—does not require the use of PCR primers that contain restriction sites

The pCR™II vector provides:
• 3'-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and Sp6 promoters for in vitro RNA transcription and sequencing
• Versatile polylinker with flanking EcoR I sites for easy excision of inserts
• M13 forward and reverse primer sites for sequencing

How TA Cloning® Works
Taq polymerase has a non-template-dependent activity that adds a single deoxyadenosine (A) to the 3' ends of PCR products. The linearized vector supplied in this kit has single 3' deoxythymidine (T) residues. This allows PCR inserts to ligate efficiently with the vector.

Kit Configurations
The TA Cloning® Kit is offered in a variety of configurations: with One Shot® INVF' Chemically Competent E. coli (K2050-01 and K2050-40), One Shot® TOP10F' Chemically Competent E. coli (K2060-01 and K2060-40), and without competent cells (K2750-20 and K2750-40) in 20- and 40-reaction kit sizes.

pYES2/CT Yeast Expression Vector (Invitrogen™)

pYES2/NT and pYES2/CT are S. cerevisiae expression vectors derived from the parental pYES2 vector. Both vectors offer the URA3 gene for selection in yeast. pYES2/NT features an N-terminal Xpress™ epitope for detection with Invitrogen's Anti-Xpress™ Antibody and a polyhistidine (6xHis) tag for purification with nickel-chelating resin. pYES2/CT contains a C-terminal V5 epitope for detection and a polyhistidine (6xHis) tag for purification.

ZeoCassette™ Vector (pCMV/Zeo) (Invitrogen™)

The ZeoCassette™ vectors allow you to incorporate the Zeocin™ resistance gene into any vector. Each vector carries a ZeoCassette™ which contains the Zeocin™ resistance marker driven by a strong constitutive promoter (Table 1). Zeocin™ is a highly effective selection agent that kills a wide variety of cell types including bacterial, yeast, plant, and mammalian cells. The ZeoCassette™ vectors can be used in a variety of ways:

• Insert your sequence of interest into one of the ZeoCassette™ vectors to create your own Zeocin™-resistant vector
• Incorporate one of the ZeoCassettes™ into a vector that youve already constructed
• Clone your promoter of choice upstream of the EM-7 promoter in pEM7/Zeo and use Zeocin™ selection in your organism of choice

Each ZeoCassette™ is flanked by large multiple cloning sites. This allows removal of the ZeoCassette™ for incorporation into your vector of choice. In addition, the Zeocin™ resistance gene in pSV40/Zeo, pCMV/Zeo, and pTEF1/Zeo is also driven by the bacterial EM-7 promoter. This allows you to use Zeocin™ to select E. colitransformants that contain the ZeoCassette™ and eliminates the need to screen colonies by restriction enzyme analysis.

pIZT/V5-His Vector Kit (Invitrogen™)

The InsectSelect™ Glow Kit features the pIZT/V5-His vector for high-level expression of your
gene of interest. This vector has the following features:

• The OpIE2 promoter for constitutive expression
• The Zeocin™ resistance gene for rapid selection of stably transfected cell lines
• C-terminal V5 epitope and polyhistidine (6xHis) sequence for detection with Invitrogen's Anti-V5 Antibody and rapid purification with nickel-chelating resin

In addition, pIZT/V5-His expresses a fusion of the green fluorescent protein and the Zeocin™ resistance
protein (Zeo-GFP). The Zeo-GFP fusion protein permits rapid selection of stably transfected cell lines
with Zeocin™ and confers a fluorescent phenotype that simplifies identification of transfected cells.

GeneArt™ CRISPR Nuclease Vector with OFP Reporter Kit (with competent cells) (Invitrogen™)

The GeneArt® CRISPR Nuclease Vector with OFP Reporter Kit is a vector system for expression of the functional components needed for CRISPR/Cas9 genome editing in mammalian cells with an orange fluorescent protein (OFP) reporter. The OFP reporter allows for fluorescence-based tracking of transfection efficiency, as well as FACS-based sorting/enrichment of Cas9 and CRISPR expressing cells. The linearized GeneArt® CRISPR nuclease vectors provide a rapid and efficient way to clone double-stranded oligonucleotides encoding a crRNA representing a desired target into an expression cassette that allows sequence-specific targeting of the Cas9 nuclease. This version of the kit includes OneShot® TOP10 Competent Cells. A version without competent cells is also available (Cat. No. A21174).

The GeneArt® CRISPR Nuclease Vector system with competent cells offers:

• Easy-to-design genome engineering system
• Affordable, ready-to-use cloning vectors
• Enrichment for hard to transfect or difficult to modify cell lines
• Competent cells included for added convenience and highest cloning efficiencies

All-in-One Vector System for CRISPR-based Genome Editing
The GeneArt® CRISPR Nuclease Vector kit offers a simple, ready-to-use, all-in-one expression vector system consisting of both a Cas9 nuclease expression cassette and a guide RNA (gRNA) cloning cassette for rapid and efficient cloning of DNA that encodes target-specific CRISPR RNA (crRNA). This system allows you to edit and engineer a genomic locus of choice in a sequence-specific manner from a single plasmid. After relevant targets have been identified with fast and easy-to-use GeneArt® CRISPR vectors, the biologically relevant mutations can be precisely created with GeneArt® Precision TALs, with high specificity and low off-target effects.

Need assistance with CRISPR gRNA design?
Our new CRISPR Search & Design tool allows you to search our database of >600,000 predesigned CRISPR gRNAs in human and mouse genes or analyze your sequence of interest for de novo gRNA designs using our proprietary algorithms. CRISPR sequences are optimized for gene knockout and target the first three transcribed exons for each gene. Search results include the top 6 CRISPR sequences with PAM sites, exon maps with gRNA binding sites, and potential off-target binding sites for each CRISPR sequence. The tool will design the correct gRNA format for your preferred CRISPR-Cas9 product, including oligos for your GeneArt® CRISPR Nuclease Vector. Start designing today >