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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.

pTracer™-SV40 Mammalian Expression Vector (Invitrogen™)

Three pTracer™ mammalian expression vectors are available that express GFP fused to the selectable marker Zeocin™ . Each vector uses a different set of promoters to express the gene of interest and the Cycle 3 GFP-Zeocin™ fusion.

All three pTracer™ vectors have the following features:

• Cycle 3 GFP-Zeocin™ fusion for selection in mammalian cell lines
• BGH polyA signal and transcription termination sequences to enhance mRNA stability

The pTracer™-SV40 vector offers:
• SV40 promoter for high-level constitutive expression of your gene of interest
• CMV promoter for high-level constitutive expression of the Cycle 3 GFP-Zeocin™ fusion
• SV40 origin for episomal replication and simple vector rescue in cell lines expressing the SV40 large T antigen

The pTracer™-CMV2 vector offers:
• CMV promoter for high-level constitutive expression of your gene of interest
• EF-1α promoter for high-level constitutive expression of the Cycle 3 GFP-Zeocin™ fusion
• Ampicillin resistance gene for selection in E. coli

The pTracer™-EF vectors offer:
• EF-1α promoter for high-level constitutive expression of your gene of interest
• CMV promoter for high-level constitutive expression of the Cycle 3 GFP-Zeocin™ fusion
• C-terminal V5 epitope tag for simple detection of recombinant fusion proteins with an Anti-V5 Antibody
• C-terminal 6xHis tag for rapid purification on nickel-chelating resin and simple detection with an Anti-His(C-term) Antibody
• Multiple cloning site supplied in three reading frames to simplify cloning in frame with the C-terminal tag

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

TA Cloning™ Kit, with pCR™2.1 Vector, without competent cells (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.

pAO815 Pichia Expression Vector (Invitrogen™)

pAO815 is a Pichia expression vector designed to clone multiple copies of a gene into a single vector. The vector contains a Bgl II site upstream of the 5´ AOX1 gene and a unique BamH I site downstream of the 3´ AOX1 transcription termination (TT) signal. Four steps are required to generate multiple copies of the gene of interest:
1. The gene is cloned into the unique EcoR I site in the vector.
2. The construct is digested with BamH I and Bgl II to release the "expression cassette" containing the AOX1 promoter, gene of interest, and 3´ AOX1 TT.
3. Concatemers of the expression cassette are generated by ligation in vitro.
4. The multiple copies are inserted back into the pAO815 vector and transformed into Pichia. This vector is included in the Multi-Copy Pichia Expression Kit.

pBAD-DEST49 Gateway™ Destination Vector (Invitrogen™)

The pBAD-DEST49 Gateway® destination vector is designed for rapid cloning with a Gateway® entry clone using lambda phage site-specific recombination and subsequent high-level, tightly regulated expression in E. coli. The pBAD-DEST49 vector features:

• The araBAD promoter for tightly regulated expression in E. coli
• HP-thioredoxin fusion for improved protein yield and solubility
• Enterokinase cleavage site for efficient cleavage of the N-terminal fusion with EKMax™
• C-terminal V5 and 6xHis tags for efficient detection and purification of fusion proteins
• R sites for efficient recombination with any attL-flanked Gateway® entry vector

pRSET-BFP Bacterial Expression Vector (Invitrogen™)

The pRSET-EmGFP, pRSET-CFP and pRSET-BFP vectors allow you to fuse a protein of interest to the widely used and well-characterized fluorescent proteins from the jellyfish Aequorea victoria (1,2) using the pRSET expression vector. These vectors contain the next-generation EGFP, Emerald Green Fluorescent Protein (EmGFP), which has been further optimized for mammalian expression, as well as Cyan (CFP) and Blue (BFP) Fluorescent Proteins for simple, noninvasive detection of recombinant protein.

Champion™ pET300/NT-DEST and pET301/CT-DEST Gateway™ Vector Kit (Invitrogen™)

The Champion™ pET300/NT-DEST and pET301/CT-DEST Gateway® Vector Kit is designed for rapid cloning with a Gateway® entry clone and subsequent high-level prokaryotic expression controlled by the strong bacteriophage T7 promoter. In addition to the T7 promoter, each vector contains only the necessary functional elements and an N- or C-terminal 6xHis tag (pET300/NT-DEST and pET301/CT-DEST, respectively) for convenient purification and detection (Figure 1). The vector kit is ideal for structural biologists who desire no or minimal modifications to their protein of interest. To maximize expression, use with MagicMedia™ E. coli Expression Medium.



Contents and Storage:
The Champion™ pET300/NT-DEST and pET301/CT-DEST Gateway® Vector Kit includes 6 µg each of pET300/NT-DEST and pET301/CT-DEST vectors and 10 µg of control vector. Store at -20“C. Guaranteed stable for 6 months when properly stored.

pcDNA™6.2-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-DEST vector offers the following key features:

•Cytomegalovirus (CMV) promoter for high-level expression
attR sites for Gateway® cloning, enabling recombination with attL-flanked fragments
•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™ RF 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.

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.

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.

pMCS minP-Tluc16-DD Vector for Luciferase Assays (Thermo Scientific™)

The Thermo Scientific™ pMCS minP-Tluc16-DD Vector is a multiple cloning site (MCS) vector designed to accept a response element sequence lacking promoter activity for study of gene regulation using the intracellular TurboLuc™16 (Tluc16) luciferase reporter.

• Intracellular Tlac16 luciferase gene, optimized for high expression in mammalian systems
• Optimized minimal core promoter (minP) and 5′ UTR region for efficient expression of Tluc16 luciferase
• Multiple cloning sites provide versatility for transfer of regulatory elements from one vector to another
• Dual-destabilization (DD) technology reduces accumulation of Tluc16 luciferase mRNA and protein in cells, enhancing the responsiveness of the assay
• Synthetic polyA terminator and Transcriptional Pause Site (TPS) included upstream of MCS to minimize non-specific transcriptional read-through

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 Tluc16 luciferase expression vector also contains dual-destabilization (DD) technology that reduces accumulation of 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 MCS to minimize non-specific transcriptional read-through. The Tluc16 luciferase activity can be measured using the Thermo Scientific TurboLuc™ One-Step Glow Assay Kit.

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

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

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pT7CFE1-NHis-GST-CHA Vector for Mammalian Cell-Free Protein Expression

pcDNA™5/FRT/TO Vector Kit (Invitrogen™)

pcDNA™FRT⁄TO vector is a 5.1 kb inducible expression vector designed for use with the Flp-In™ T-REx™ System. The control plasmid, pcDNA™5⁄FRT⁄TO⁄CAT, is also included for use as a positive control for transfection and expression in your Flp-In™ T-REx™ host cell line of choice.

The pcDNA™5⁄FRT⁄TO vector contains the following elements:

• A hybrid human cytomegalovirus (CMV)⁄TetO2 promoter for high-level, tetracycline-regulated expression of the gene of interest in a wide range of mammalian cells
• Multiple cloning site with 10 unique restriction sites to facilitate cloning the gene of interest
• FLP Recombination Target (FRT) site for Flp recombinase-mediated integration of the vector into the Flp-In™ T-REx™ host cell line
• Hygromycin resistance gene for selection of stable cell lines

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

Thermo Scientific pT7CFE1-NHA 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-NHA Vector is available with single affinity HA tag at the N-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

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pT7CFE1-NHis-GST-CHA Vector for Mammalian Cell-Free Protein Expression

Vivid Colors™ pcDNA™6.2/N-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

Gateway™ pENTR™ 4 Dual Selection Vector (Invitrogen™)

Gateway® entry vectors are designed to clone DNA sequences using restriction endonucleases and ligase to create a Gateway® entry clone. The resulting entry clone is ready for recombination with a destination vector to create an expression clone.

The Gateway® entry vectors (Table 1) offer the following:
• attL1 and attL2 sites for site-specific recombination of the entry clone with a Gateway® destination vector to ensure cloning of the gene of interest in the correct orientation for expression
• Kozak consensus sequence for efficient translation initiation in eukaryotic systems
• Ribosome binding site for efficient translation initiation in prokaryotic systems (pENTR™ 1A Dual Selection, pENTR™3C Dual Selection, and pENTR™11 Dual Selection vectors only)
• rrnB transcription termination sequences to prevent basal expression of the PCR product of interest in E. coli
• pUC origin for high-copy replication and maintenance of the plasmid in E. coli
• Kanamycin resistance gene for selection in E. coli
• The ccdB⁄chloramphenicol fusion gene located between the two attL sites for
o negative selection and
o Chloramphenicol selection in E. coli
• Kanamycin resistance gene for selection in E. coli

pMCS Tluc16-DD Hygro Vector for Luciferase Assays (Thermo Scientific™)

The Thermo Scientific™ pMCS Tluc16-DD Hygro Vector is a multiple cloning site (MCS) vector designed to accept a promoter sequence for study of gene regulation using the intracellular TurboLuc™16 (Tluc16) luciferase reporter.

• Intracellular Tluc16 luciferase gene, optimized for high expression in mammalian systems
• Multiple cloning sites provide versatility for transfer of regulatory elements from one vector to another
• Dual-destabilization (DD) technology reduces accumulation of Tluc16 luciferase mRNA and protein in cells, enhancing the responsiveness of the assay
• Synthetic polyA terminator and Transcriptional Pause Site (TPS) included upstream of MCS to minimize non-specific transcriptional read-through
• Hygromycin resistance gene for use as selection marker to generate mammalian stable cell lines

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 pMCS Tluc16-DD Hygro 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 MCS to minimize non-specific transcriptional read-through. This vector also contains the hygromycin resistance gene, which can be used as a mammalian selection marker to generate stable cell lines. The Tluc16 luciferase activity can be measured using the Thermo Scientific TurboLuc™ One-Step Glow Assay Kit.

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.

pCMV-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 CMV (Cytomegalovirus) 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 pCMV-Green Renilla Luc vector:

• Strong (CMV) 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

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

Thermo Scientific pCMV-Red Firefly Luc is a constitutive expression vector having the luciferase gene under the CMV (Cytomegalovirus) 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 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

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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

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.

TA Cloning™ Kit, Dual Promoter, with pCR™II Vector, without competent cells (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.

Gateway™ pDEST™8 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.

pRSET-CFP Bacterial Expression Vector (Invitrogen™)

The pRSET-EmGFP, pRSET-CFP and pRSET-BFP vectors allow you to fuse a protein of interest to the widely used and well-characterized fluorescent proteins from the jellyfish Aequorea victoria (1,2) using the pRSET expression vector. These vectors contain the next-generation EGFP, Emerald Green Fluorescent Protein (EmGFP), which has been further optimized for mammalian expression, as well as Cyan (CFP) and Blue (BFP) Fluorescent Proteins for simple, noninvasive detection of recombinant protein.

Gateway™ pcDNA™-DEST47 Vector (Invitrogen™)

The pcDNA™ vectors are designed for high-level, constitutive expression in a variety of mammalian cell lines. The Gateway® pcDNA™-DEST47 vector offers the following key features:

•C-terminal Cycle 3 GFP tag for rapid detection of recombinant protein
•Cytomegalovirus (CMV) promoter for high-level expression
attR sites for Gateway® cloning, enabling recombination with attL-flanked fragments
•Neomycin resistance gene for 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.

pEF5/FRT/V5-DEST™ 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.