Shop All DNA Vectors

pJTI™ R4 CMV-TO MCS pA Vector

The pJTI™ R4 CMV-TO MCS pA vector is designed for the expression of your gene of interest under the control of the tet-inducible CMV promoter after restriction enzyme cloning and retargeting into the genomic R4 site of a Jump-In™ parental cell line.

This vector can be used for inducible or constitutive expression of your gene of interest, depending on which Jump-In™ parental cell line you use. When this vector is retargeted into a Jump-In™ T-REx™ parental cell line, gene expression is controlled by the tet-operon and can be induced by adding doxycycline to the growth media. When used with a Jump-In™ parental cell line such as the Jump-In™ GripTite™ HEK293 cell line, constitutive expression is achieved after retargeting. The R4 sites in the Jump-In™ parental cell lines result in a high retargeting efficiency, requiring less effort and less time than traditional cell engineering methods. Retargeting of Jump-In™ parental cell lines results in creation of an isogenic pool that is sufficient for cell-based experiments without the need for clonal selection. Alternatively, the high retargeting efficiency allows for easy selection of a positive stable clone for expressing your gene of interest.

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.

Gateway™ pMT-DEST48 Vector (Invitrogen™)

The pMT-DEST48 destination vector is designed for rapid cloning with a Gateway® entry clone using lambda phage site-specific recombination and subsequent expression in Drosophila S2 cells. As part of the DES® Expression System, pMT-DEST48 uses the Drosophila metallothionein gene promoter that is induced in S2 cells upon addition of copper sulfate or cadmium chloride to the culture medium. The pMT-DEST48 vector offers the following features:

• C-terminal V5 epitope tag for rapid detection with Invitrogens Anti-V5 Antibody
• C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin
• R sites for efficient recombination with any attL-flanked Gateway® entry vector

pcDNA™3.1/V5-His TOPO™ TA Expression Kit (Invitrogen™)

The pcDNA™3.1/V5-His TOPO® TA Expression Kit offers one-step cloning of Taq-amplified PCR products into a high-level expression vector. Topoisomerase activation of the pcDNA3.1/V5-His-TOPO® vector allows PCR products to be ligated in just 5 minutes on your bench top and results in 90% recombinants.

In addition, the vector includes the following features:

• Strong CMV promoter for high-level, constitutive expression.
• C-terminal V5 epitope tag for efficient detection of recombinant proteins with an Anti-V5 antibody.
• C-terminal polyhistidine (6xHis) sequence for purification using nickel-chelating resin and detection with an Anti-His (C-term) antibody.

TrueTag™ Donor DNA Kit, GFP (Invitrogen™)

The Invitrogen TrueTag Donor DNA Kit, GFP, provides a complete and rapid solution to produce transfection-ready donor DNA to drive high-efficiency gene tagging in genome editing experiments. Benefits of the TrueTag Donor DNA kits include:
• Simple PCR amplification to add homology arms to the kit-provided donor template
• Reduces time needed for cloning of donor templates from days to hours
• Same-day transfection-ready donor DNA
• Obtain up to 100% edited cells
• Works with CRISPR, TALENs, and zinc-finger nucleases

The TrueTag Donor DNA Kit, GFP, provides everything you need to produce a high-quality donor DNA to tag your gene with GFP—simply provide two oligos to target the gene. This comprehensive yet flexible kit enables tagging of the carboxy- (C-) or amino- (N-) terminus of the gene with a choice of puromycin or blasticidin resistance cassettes to drive up to 100% edited cells. Validated with our TrueCut Cas9 Protein v2 and TrueGuide Synthetic gRNAs, the TrueTag Donor DNA kits are compatible with current genome editing technologies, including CRISPR, TALENs, and zinc-finger nucleases.

Each TrueTag Donor DNA Kit includes:
• Four linear donor templates: N-terminus/puromycin resistance, N-terminus/blasticidin resistance, C-terminus/puromycin resistance, C-terminus/blasticidin resistance
• Phusion Flash High-Fidelity PCR Master Mix
• PCR cleanup columns and buffers
• Positive control primers to tag human beta-actin (ACTB)

Gateway™ pDEST™27 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.

TOPO™ TA Cloning™ Kit for Subcloning, with One Shot™ Mach1™ T1 Phage-Resistant Chemically Competent E. coli (Invitrogen™)

TOPO® TA Cloning® Kits for Subcloning provide a highly efficient, 5-minute, one-step cloning strategy ("TOPO® cloning") for the direct insertion of Taq polymerase–amplified PCR products into a plasmid vector for subcloning. Each kit uses the pCR™ 2.1-TOPO® TA vector with convenient restriction sites for subcloning. The cloning kits are available with a variety of competent cells, or without competent cells, depending upon your needs and budget. Features of the TOPO® TA Cloning® Kits for Subcloning:

Fast and easy—go from PCR to clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—obtain up to 95% clones with correct insert
Proven—reliable performance for over a decade with over 4,000 citations
Simple—no ligase, post-PCR procedures, or PCR primers containing specific sequences are required

TOPO® TA Cloning® Kits for Subcloning—overview

Vector: pCR™ 2.1-TOPO® TA vector—subcloning vector with 15 convenient restriction sites flanking your insert for easy, directional subcloning

Cloning method: TOPO® TA Cloning® —Topoisomerase I–based, 5-minute ligation of PCR products with 3′-A overhangs (Taq polymerase amplified) to TOPO® vector with T overhangs

Competent cells: various options—choose from kits with either general, high-efficiency, bacteriophage T1–resistant, fast-growing competent cells, or use your own

pCR™ 2.1-TOPO® TA vector—cloning and subcloning simplicity
The pCR™ 2.1-TOPO® TA vector is linearized with 3'-thymidine (T) overhangs for direct ligation of Taq polymerase–amplified PCR products (TA cloning®) and is "activated" with covalently bound Topoisomerase I. There is no need to add ligase and cloning is complete in 5 minutes. EcoRI sites flanking the PCR product insertion site allow for easy excision of inserts or use any combination of 15 convenient restriction sites flanking your PCR insert for easy, directional subcloning.

pCR™ 2.1-TOPO® TA clone selection and manipulation
The pCR™ 2.1-TOPO® TA vector contains both ampicillin and kanamycin resistance markers and the LacZα gene for blue/white screening.

Simplified TOPO® -based cloning
Using TOPO® cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transform E. coli competent cells.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen less clones, saving time and money. The pCR™ 2.1-TOPO® TA vector used in this kit comes with 3'-T overhangs for efficient ligation of Taq polymerase–amplified PCR products, which contain 3'-A overhangs.

The standard in cloning
When it comes to cloning, TOPO® cloning technology has been a reliable partner for thousands of scientists for over ten years. Fast, simple-to-use, and efficient, TOPO® cloning has been applied to many different vectors for a wide array of applications.

TOPO® TA Cloning® Kits for Subcloning—kit options
The TOPO® TA Cloning® Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:

• General cloning: TOP10 cells (Cat. No. K4500-01, K4500-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K4560-01, K4560-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R cells (Cat. No. K4520-01, K4520-40)
• Fast growth: Mach1™ -T1R Chemically Competent E. Coli (Cat. No. K4510-20)
• Provide your own: for flexibility and to save money (Cat. No. 450641)

We also offer two versions of the kit that include a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K4500-02 and K4510-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.

pcDNA™3.1 (+) Mammalian Expression Vector (Invitrogen™)

This pcDNA™3.1(+) vector is designed for high-level, constitutive expression in a variety of mammalian cell lines. It contains a Geneticin® selectable marker and a forward-orientation multiple cloning site.

The pcDNA™3.1 Expression Vector Family
Three untagged versions of pcDNA™3.1 (available separately), each with a different selectable marker (Geneticin®, Zeocin™, or Hygromycin), are for use alone or in co-transfections. All three vectors offer the following features:

• Cytomegalovirus (CMV) enhancer-promoter for high-level expression
• Large multiple cloning site in either forward (+) or reverse (-) orientations
• Bovine Growth Hormone (BGH) polyadenylation signal and transcription termination sequence for enhanced mRNA stability
• SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen (i.e., COS-1 and COS-7)
• Ampicillin resistance gene and pUC origin for selection and maintenance in E. coli

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

Vivid Colors™ pcDNA™6.2/C-EmGFP-GW/TOPO™ Mammalian Expression Vector (Invitrogen™)

The Vivid Colors™ pcDNA™6.2 Fluorescent Protein TOPO® Expression Vectors (Figure 1) allow you to rapidly clone your gene and fuse it to the widely used and well characterized Fluorescent Proteins (FPs) from the jellyfish Aequorea victoria (1, 2). These powerful TOPO® cloning vectors contain the Emerald Green Fluorescent Protein (EmGFP) or the Yellow Fluorescent Protein (YFP) for simple, non-invasive detection of recombinant protein (Figure 2). Both FPs have been humanized for optimal mammalian expression (3). The Vivid Colors™ pcDNA™6.2 Fluorescent Protein TOPO® Expression Vectors offer:
• Topoisomerase I for one-step, 5-minute TOPO® cloning of your PCR-amplified gene of interest
• CMV promoter for high-level expression of the recombinant fluorescent fusion protein
• Ability to fuse EmGFP or YFP to the N- or C-terminus of your protein
• Bsd resistance marker for rapid selection of stable cell lines

pYES2/NT A, B, & C Yeast Expression Vectors (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.

NT-GFP Fusion TOPO™ Expression Kit (Invitrogen™)

The GFP Fusion TOPO® TA Expression Kits are designed to allow fusion of a protein of interest to the Cycle 3 GFP protein. These kits provide topoisomerase-I activated pcDNA3.1/NT-GFP-TOPO® or pcDNA3.1/CT-GFP-TOPO® vectors (Figure 1). Taq-amplified DNA fragments are ligated into these vectors in a simple 5-minute reaction right on your bench top. The vectors are designed for high-level expression of transient or stable GFP fusion proteins in a wide range of mammalian cells (Figure 2). Expression is easily detected in living cells using fluorescence. In addition, recombinant proteins expressed from these vectors can be detected on western blots using GFP Antiserum.

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

Thermo Scientific pT7CFE1-CHA 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-CHA Vector is available with single affinity HA 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-NHA 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

TrueTag™ Donor DNA Kit, RFP (Invitrogen™)

The Invitrogen TrueTag Donor DNA Kit, RFP, provides a complete and rapid solution to produce transfection-ready donor DNA to drive high-efficiency gene tagging in genome editing experiments. Benefits of the TrueTag Donor DNA kits include:
• Simple PCR amplification to add homology arms to the kit-provided donor template
• Reduces time needed for cloning of donor templates from days to hours
• Same-day transfection-ready donor DNA
• Obtain up to 100% edited cells
• Works with CRISPR, TALENs, and zinc-finger nucleases

The TrueTag Donor DNA Kit, RFP, provides everything you need to produce a high-quality donor DNA to tag your gene with RFP—simply provide two oligos to target the gene. This comprehensive yet flexible kit enables tagging of the carboxy- (C-) or amino- (N-) terminus of the gene with a choice of puromycin or blasticidin resistance cassettes to drive up to 100% edited cells. Validated with our TrueCut Cas9 Protein v2 and TrueGuide Synthetic gRNAs, the TrueTag Donor DNA kits are compatible with current genome editing technologies, including CRISPR, TALENs, and zinc-finger nucleases.

Each TrueTag Donor DNA Kit includes:
• Four linear donor templates: N-terminus/puromycin resistance, N-terminus/blasticidin resistance, C-terminus/puromycin resistance, C-terminus/blasticidin resistance
• Phusion Flash High-Fidelity PCR Master Mix
• PCR cleanup columns and buffers
• Positive control primers to tag human beta-actin (ACTB)

pcDNA™6.2/GW/D-TOPO™ Expression Kit (Invitrogen™)

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

•Cytomegalovirus (CMV) promoter for high-level expression
•Adapted for Directional TOPO® Cloning, enabling you to use proofreading polymerases and to clone your PCR products in a specific orientation
•Blasticidin resistance gene for efficient stable selection
•C-terminal V5 tag for easy detection
•Ampicillin resistance gene and pUC origin for selection and maintenance in E. coli

TOPO® Cloning
Using restriction enzymes to clone your gene into an expression vector often forces you to compromise the final sequence of your insert (Figure 1A), especially when there are no useful restriction sites close to your genes coding sequence. This may result in suboptimal spacing of expression elements or incorporation of non-native amino acid residues, which can reduce your expression levels and/or cause the production of non-functional protein.

In addition to being a more effective way to clone, TOPO® Cloning eliminates these potential expression problems. TOPO® Expression Vectors enable you to insert the exact DNA sequence you require simply by performing PCR with appropriately designed primers. Your PCR product is cloned at a high efficiency in only five minutes into a topoisomerase I-activated expression vector. The resulting recombinant expression vector contains your exact DNA sequence without any non-coding regions (Figure 1B).

Many of our powerful expression vectors are available adapted for one-step TOPO® cloning and expression of PCR products. In addition, several expression vectors are now adapted for Directional TOPO® Cloning, enabling you to use proofreading polymerases and to clone your PCR products in a specific orientation.