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TC-FlAsH™ TC-ReAsH™ II In-Cell Tetracysteine Tag Detection Kit, with Mammalian TC-Tag Gateway™ Expression Vectors (Green Fluorescence) (Red Fluorescence) (Invitrogen™)

TC-FlAsH™ TC-ReAsH™ II In-Cell Tetracysteine Tag Detection Kit is an expression tag-based fluorescence labeling system for live-cell labeling. The expression construct is created when the Gateway entry clone (containing the gene of interest) is recombined with one of two destination vectors (pcDNA6.2/nTC-Tag-DEST for N-terminal tagging or pcDNA6.2/cTC-Tag-DEST for C-terminal tagging). The expression construct is then used to transform the host strain. The protein can be detected either with green-fluorescent FlAsH-EDT2 reagent or red-fluorescent ReAsH-EDT2 reagent. The tag is very small (6 amino acids) and the protein of interest is fluorescent only when the labeling reagent is added. BAL wash buffer replaces the previously supplied Disperse Blue 3 and EDT-based wash buffer as an olfactorily more agreeable reagent that yields superior signal to noise.

The kit contains FlAsH-EDT2 labeling reagent, ReAsH-EDT2 labeling reagent, BAL wash buffer, pcDNA6.2/cTC-Tag-DEST, pcDNA6.2/nTC-Tag-DEST, and pcDNA6.2/nTC-Tag-p64 Control Plasmid. BAL wash buffer should be stored at 4°C, and the rest of the kit stored at -20°C, protected from light. The kit is stable for 6 months when stored correctly.

Expressway™Lumio™ Expression and Detection System, without vector (Invitrogen™)

The Expressway™ Lumio™ Expression and Detection System takes advantage of the Lumio™ recognition sequence, a small, six amino acid sequence (Cys-Cys-Pro-Gly-Cys-Cys). The Lumio™ detection reagent binds the recognition sequence with high specificity and affinity, resulting in a bright fluorescent signal for real-time protein production analysis and immediate in-gel protein detection. In addition, Expressway’s specialized E. coli lysate, derived from a slyD mutant, eliminates nonspecific binding of the Lumio™ Green Detection Reagent to the endogenous SlyD protein and provides optimal background for detection of recombinant proteins (Figure 1). The Lumio™ Green Detection Kit is included in the system for real-time detection of protein synthesis as well as easy in-gel protein detection. The Lumio™ vectors (Figures 2 and 3) also feature:

attR sites for efficient recombination with any attL-flanked Gateway® entry vector
N-terminal Lumio™ tag (pEXP3-DEST vector) with TEV cleavage site for efficient removal of the Lumio™ sequence following purification
C-terminal Lumio™ tag (pEXP4-DEST vector) for easy, and immediate in-gel detection
T7 promoter, ribosome binding site, and T7 terminator optimally spaced for cell-free protein expression

pEF6/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pcDNA™6/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pEF6/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pTrcHis A, B, & C Bacterial Expression Vectors (Invitrogen™)

Our pTrcHis A, B, & C vectors are designed to offer enhanced translation initiation and high-level expression in E. coli. These vectors feature:

• High-level regulated transcription from the trc promoter
• Enhanced translation efficiency of eukaryotic genes in E. coli
• The lacO operator and lacIq repressor gene for transcriptional regulation in any E. coli strain

This particular vector offers:

• N-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an anti-HisG antibody
• N-terminal Xpress™ epitope for easy detection with an anti-Xpress™ antibody
• Enterokinase cleavage site for removal of fusion tag

For C-terminal polyhistidine tag and c-myc epitope, please see our pTrcHis2 A, B, & C Vector.

pcDNA™4/TO/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

A Tetracycline-Regulated Expression System without Viral Transactivators
The T-REx™ System yields higher levels of induced expression than any other regulated mammalian expression system. It utilizes the complete CMV promoter and adds control elements from the bacterial tetracycline resistance operon to effectively repress and derepress transcription from one of the strongest mammalian promoter sequences known (1,2).

Specific Activation
The T-REx™ System uses a repressor mechanism that blocks transcription from the powerful CMV promoter in the absence of tetracycline. Because the T-REx™ System elements do not use viral transactivators, you can achieve high-level expression from the complete CMV promoter without secondary, non-specific activation of host genes.

The T-REx™ Mechanism
The T-REx™ transcriptional control elements are illustrated in Figure 1. Two tetracycline operator sequences (TetO2) have been inserted between the TATA box of the CMV promoter and the transcriptional start site. The TetO2 sequence itself has no effect on expression. When the tetracycline repressor protein (TR) is present, it effectively binds the TetO2 sites and blocks transcription initiation. Tetracycline added to the culture medium binds to, and changes the conformation of, the TR protein. This change causes the TR protein to release the TetO2 sites, derepressing transcription from the CMV promoter. The result is high-level expression of the gene of interest (Figure 2). Expression levels can be modulated based on the tetracycline concentration and can be induced to levels that are achieved with constitutive CMV expression vectors.
T-REx™ is a powerful inducible mammalian expression system that allows you to regulate expression from the complete human cytomegalovirus (CMV) enhancer-promoter. T-REx™inducible expression vectors offer the following features:

• Complete CMV enhancer-promoter sequence containing two copies of the tetracycline operator TetO2 sequence for high-level regulated expression
• Zeocin™ or hygromycin resistance gene for effective selection of stable mammalian cell lines
• Large multiple cloning site to simplify cloning

In addition, pcDNA™4/TO/myc-His offers a c-myc epitope for rapid detection of the recombinant protein with an Anti-myc Antibody and a polyhistidine (6xHis) sequence for simple purification of the recombinant protein with nickel-chelating resin and detection with Anti- His(C-term) Antibody.

The regulatory vector, pcDNA™6/TR, is provided for high-level expression of the tetracycline repressor (TR) protein. This vector expresses the Blasticidin resistance gene for rapid selection of mammalian cell lines that stably express the TR protein.

pcDNA™3.1/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pcDNA™6/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pcDNA™4/HisMax A, B, & C Mammalian Expression Vectors (Invitrogen™)

pcDNA™4/HisMax is specifically designed to maximize protein expression in a variety of mammalian cells. The vector contains the QBI SP163 translational enhancer to increase expression levels two- to five-fold above those seen with the promoter alone (Figure 1). In addition to SP163-enhanced expression, pcDNA™4/HisMax includes a cleavable N-terminal Xpress™ tag for rapid detection of recombinant protein with an Anti-Xpress™ Antibody (Figure 2). pcDNA™4/HisMax is available TOPO® Cloning-ready for five-minute cloning of Taqamplified PCR products.

pTrcHis2 A, B, & C Bacterial Expression Vectors (Invitrogen™)

Our pTrcHis A, B, & C vectors are designed to offer enhanced translation initiation and high-level expression in E. coli. These vectors feature:

• High-level regulated transcription from the trc promoter
• Enhanced translation efficiency of eukaryotic genes in E. coli
• The lacO operator and lacIq repressor gene for transcriptional regulation in any E. coli strain

This particular vector offers:

• C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an anti-His(C-term) antibody
• C-terminal c-myc epitope for easy detection of fusion proteins with an anti-myc antibody

For N-terminal polyhistidine tag and Xpress™ epitope, please see our pTrcHis A, B, & C Vector.

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.

pPICZ A, B, & C Pichia Vectors (Invitrogen™)

The pPICZ A, B, & C Pichia Vectors are designed for simple cloning and selection, high-level expression, and rapid detection and purification of the recombinant protein. These vectors contain the Zeocin™ resistance gene for direct selection of multi-copy integrant strains. By selecting with increasing amounts of Zeocin™, strains with multiple copies of your gene of interest integrated into the genome are obtained. Increasing the number of copies of the gene of interest in a recombinant Pichia strain can result in higher expression levels. The vectors are included in the EasySelect™ Pichia Expression Kit (Cat. No. K1740-01).

Features of the pPICZ vectors include:

• Inducible AOX1 promoter for high-level expression in Pichia pastoris
c-
myc epitope tag for convenient detection with an Anti-myc Antibody
• C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• Zeocin™ resistance for direct selection of multi-copy integrants

Mammalian Lumio™ Gateway™ Vectors, with Lumio™ Green In-Cell Detection 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.

pGAPZα A, B, & C Pichia pastoris Expression Vectors (Invitrogen™)

pGAPZ A, B, & C and pGAPZ A, B, & C are expression vectors designed for high-level, constitutive expression in Pichia pastoris. pGAPZ and pGAPZ were created by replacing the methanol-regulated AOX1 promoter with the constitutive, glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter in the backbone of the pPICZ vectors. Although the yield
of any protein constitutively expressed in the Pichia system is dependent on the toxicity of the protein to yeast, constitutive expression under the control of pGAPZ or pGAPZ vectors can sometimes produce greater yields than inducible expression (1).

pEF4/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pGAPZ A, B, & C Pichia pastoris Expression Vectors (Invitrogen™)

pGAPZ A, B, & C and pGAPZ A, B, & C are expression vectors designed for high-level, constitutive expression in Pichia pastoris. pGAPZ and pGAPZ were created by replacing the methanol-regulated AOX1 promoter with the constitutive, glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter in the backbone of the pPICZ vectors. Although the yield
of any protein constitutively expressed in the Pichia system is dependent on the toxicity of the protein to yeast, constitutive expression under the control of pGAPZ or pGAPZ vectors can sometimes produce greater yields than inducible expression (1).

pcDNA™4/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pRSET A, B, & C Bacterial Expression Vectors (Invitrogen™)

The pRSET vector is designed for high-level prokaryotic expression controlled by the strong bacteriophage T7 promoter. Expression is induced by the production of T7 RNA polymerase in BL21(DE3) E. coli. These cells also produce T7 lysozyme to reduce basal expression of target genes. The pRSET vector offers:

• Bacteriophage T7 promoter for high-level expression
• T7 gene 10 sequence to provide protein stability
• N-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-HisG Antibody
• N-terminal Xpress™ epitope for detection with the Anti-Xpress™ Antibody
• Enterokinase cleavage site for removal of fusion tag

A set of three vectors is provided (A, B, and C). Each has the N-terminal tag coding sequence in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.

Episomal iPSC Reprogramming Vectors (Invitrogen™)

Episomal iPSC Reprogramming Vectors are a cost-effective mixture of three vectors designed to provide the optimal system for generating transgene-free and virus-free induced pluripotent stem cells (iPSCs) in a feeder-free environment. Originally developed by Junying Yu and James Thomson(1) and further optimized by Cellular Dynamics International, these Episomal iPSC Reprogramming Vectors have proven successful in reprogramming a number of different somatic cell types.
• Safe for all stages of your iPSC research—transgene-free and viral-free reprogramming allows use from basic to pre-clinical research
• Reprogram a variety of somatic cell types—provides flexibility in somatic cell selection
• Optimized for feeder-free reprogramming—allows for defined and feeder-free reprogramming when used with Essential 8™ Medium

Create Transgene- and Virus-free iPSCs
The Episomal iPSC Reprogramming Vectors are a well-described system for producing transgene-free, virus-free iPSCs, providing a source of iPSCs for all stages of your pluripotent stem cell research. Other reprogramming methods, such as lentivirus, contain transgenes that can integrate into the host genome, potentially disrupting the genome or causing unpredictable results. These episomal vectors are introduced into the cell by electroporation. As oriP/EBNA1 vectors, they contain all 6 reprogramming factors (Oct4, Sox2, Nanog, Lin28, Klf4 and lMyc) and replicate extrachromosomally only once per cell cycle. At this replication rate, the episomes are lost at a rate of approximately 5% per cell generation.

Generate iPSCs from a Wide Variety of Somatic Cell Types
iPSCs have been generated with episomal vectors from a range of somatic cells including fibroblasts, bone marrow mononuclear cells, PBMCs, lymphoblast B cells, and various disease-type fibroblasts and PBMCs. Each kit provides enough material for 6 reprogramming experiments.

Optimized for Feeder-free Reprogramming with Essential 8™ Medium
The Episomal iPSC Reprogramming Vectors were designed in the laboratories of James Thomson and Cellular Dynamics International for use with Essential 8™ Medium, thus providing an optimal environment for defined, feeder-free reprogramming.

Commercialized in Partnership with Cellular Dynamics International.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

Reference:
1. Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences; Yu J, Hu K, Smuga-Otto K, Tian S, Stewart R, Slukvin II, Thomson J. Science. 2009; 324:797-801.

pcDNA™3.1/His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pcDNA™3.1(-)/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pEF1/His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pPICZα A, B, & C Pichia Vectors (Invitrogen™)

pPICα A, B, and C vectors are 3.6 kb vectors used to express and secrete recombinant proteins in Pichia pastoris. Recombinant proteins are expressed as fusions to an N-terminal peptide encoding the Saccharomyces cerevisiae á-factor secretion signal. These vectors allow high-level, methanol inducible expression of the gene of interest in Pichia, and can be used in any Pichia strain including X-33, SMD1168H, and KM71H. pPICα vectors contain the following elements:


• Contains AOX1 promoter for tightly regulated, methanol-induced expression of the gene of interest
• All three reading frames (A, B, C versions) are provided to facilitate in-frame cloning with the C-terminal peptide
• α-factor secretion signal for directing secreted expression of the recombinant protein
• Zeocin resistance gene for selection in both E. coli and Pichia
• C-terminal peptide containing the c-myc epitope and a polyhistidine (6xHis) tag for detection and purification of a recombinant fusion protein

pMT/BiP/V5-His A, B, & C Drosophila Expression Vectors (Invitrogen™)

The DES®-Inducible/Secreted Kit provides the vector pMT/BiP/V5-His for inducible, secreted expression of recombinant proteins. This vector offers the inducible metallothionein promoter that is induced upon addition of copper sulfate or cadmium chloride. The N-terminal signal sequence from the insect BiP gene is provided to direct the recombinant fusion protein through the secretory pathway of S2 cells into the culture medium. The pMT/BiP/V5-His vector offers the following additional features:

• Small size (3.6 kb) to improve DNA yields and increase subcloning efficiency
• 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

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the BiP signal sequence.

pcDNA™3.1/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pcDNA™4/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pPIC6α A, B, & C Pichia Vectors (Invitrogen™)

pPIC6 A, B, & C are Pichia pastoris expression vectors designed for simple selection,
high-level expression, and rapid protein purification and detection. The pPIC6 A,
B, & C vectors are designed for high-level secreted expression. Both sets of vectors
have the following features:
Blasticidin resistance for direct selection of multi-copy integrants
Inducible AOX1 promoter for high-level expression in Pichia pastoris
C-terminal c-myc epitope for convenient detection with an Anti-myc Antibody
C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating
resin and detection with an Anti-His(C-term) Antibody
pPIC6 also features:
An É -factor secretion signal for efficient transport of proteins to the medium

pUB6/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pEF1/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pMT/V5-His A, B, & C Drosophila Expression Vectors (Invitrogen™)

The DES®-Inducible Kit provides the expression vector pMT/V5-His for expression of recombinant proteins. This vector 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/V5-His vector offers the following features:

• Small size (3.5 kb) to improve DNA yields and increase subcloning efficiency
• C-terminal V5 epitope tag for rapid detection with Invitrogen's Anti-V5 Antibody
• C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the coding sequence of the C-terminal tag.

pPIC6 A, B, & C Pichia Vectors (Invitrogen™)

pPIC6 A, B, & C are Pichia pastoris expression vectors designed for simple selection,
high-level expression, and rapid protein purification and detection. The pPIC6 A,
B, & C vectors are designed for high-level secreted expression. Both sets of vectors
have the following features:
Blasticidin resistance for direct selection of multi-copy integrants
Inducible AOX1 promoter for high-level expression in Pichia pastoris
C-terminal c-myc epitope for convenient detection with an Anti-myc Antibody
C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating
resin and detection with an Anti-His(C-term) Antibody
pPIC6 also features:
An É -factor secretion signal for efficient transport of proteins to the medium

pEF1/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pAc5.1/V5-His A, B, & C Vectors (Invitrogen™)

pAC5.1/V5-His vectors are designed for use in Drosophila cells to achieve high-level transient expression of recombinant proteins. The vector offers the strong, constitutive promoter from the Drosophila actin 5C gene. The pAC5.1/V5-His vectors can be used with the DES™-Inducible Kits (K512001 and K412001) for constitutive expression of your protein of interest. The pAc5.1/V5-His vectors also offer the following features:

• Small size (5.4 kb) to improve DNA yields and increase subcloning efficiency
• 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

To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the coding sequence of the C-terminal tag.

pSecTag2/Hygro A, B, & C Mammalian Expression Vectors (Invitrogen™)

pSecTag2 and pSecTag2/Hygro are mammalian expression vectors designed for the secretion, purification, and detection of fusion proteins. Each vector has a large multiple cloning site in three reading frames to simplify cloning in frame with the N-terminal secretion signal. The vectors (Figure 1) offer the following features:

• Secretion signal from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Figure 2)
• Cytomegalovirus (CMV) promoter for high-level constitutive expression
• C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• C-terminal c-myc epitope for detection with an Anti-myc Antibody
• Bovine Growth Hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability
• SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen (e.g., COS-1, COS-7)

The pSecTag2 vectors carry the Zeocin™ resistance gene for cost-effective selection in mammalian cells. Zeocin™ selection can also be used in E. coli.

The pSecTag2/Hygro vectors have the Hygromycin B resistance gene for selection of stable mammalian cell lines.

pSecTag2 A, B, & C Mammalian Expression Vectors (Invitrogen™)

pSecTag2 and pSecTag2/Hygro are mammalian expression vectors designed for the secretion, purification, and detection of fusion proteins. Each vector has a large multiple cloning site in three reading frames to simplify cloning in frame with the N-terminal secretion signal. The vectors (Figure 1) offer the following features:

• Secretion signal from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Figure 2)
• Cytomegalovirus (CMV) promoter for high-level constitutive expression
• C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• C-terminal c-myc epitope for detection with an Anti-myc Antibody
• Bovine Growth Hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability
• SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen (e.g., COS-1, COS-7)

The pSecTag2 vectors carry the Zeocin™ resistance gene for cost-effective selection in mammalian cells. Zeocin™ selection can also be used in E. coli.

The pSecTag2/Hygro vectors have the Hygromycin B resistance gene for selection of stable mammalian cell lines.

pTracer™-EF A, B, & C Mammalian Expression Vectors (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

ViraPower™ HiPerform™ Promoterless Gateway™ Vector Kit (Invitrogen™)

The ViraPower™ HiPerform™ Promoterless Gateway® Vector Kit contains the MultiSite Gateway®-adapted ViraPower™ HiPerform™ promoterless lentiviral expression vector, pLenti6.4⁄R4R2⁄V5-DEST™ for easy recombination-based cloning and lentiviral-based high-level expression of a target gene from any promoter of choice in dividing and non-dividing mammalian cells. The pLenti6.4⁄R4R2⁄V5-DEST™ vector is equipped with two key genetic elements, making it a HiPerform™ vector: the Woodchuck Posttranscriptional Regulatory Element (WPRE) and the central Polypurine Tract (cPPT) sequence from the HIV-1 integrase gene to produce at least 4-fold increase in protein expression compared to vectors lacking these elements.

Advantages
• Generates replication-incompetent lentivirus for transducing dividing and non-dividing mammalian cells
• Easy, simultaneous, recombination-based cloning of multiple DNA fragments in a defined order and orientation using MultiSite Gateway® technology
• Expression of the target gene under the control of a promoter of choice
• Stable, long-term expression
• Enhanced protein expression, up to 4-fold or greater, compared to traditional lentiviral expression systems

Key Features
• WPRE from the woodchuck hepatitis virus, increases transgene expression and cPPT from the HIV-1 integrase gene, increases the copy number of lentivirus integrating into the host genome, thus increasing viral titer. WPRE and cPPT together produce at least a four-fold increase in protein expression in most cell types, compared to other vectors that do not contain these elements.
• Promoterless vector to express the target gene under the control of a promoter of choice
• Blasticidin selection marker for stable selection under control of PGK promoter for long-term, persistent expression

Kit includes
• pLenti6.4⁄R4R2⁄V5-DEST™ Kit
• pENTR™ 5’ TOPO® TA Cloning Kit (Cat # K59120)
• One Shot® Stbl3™ Chemically Competent E. coli (Cat # C737303)
• pENTR® Gus positive control plasmid

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

pcDNA™6/V5-His Mammalian Expression Vector, with 50 mg blasticidin (Invitrogen™)

All pcDNA™ vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

TA Cloning™ Kit, with pCR™2.1 Vector and One Shot™ TOP10F' 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.

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.

pcDNA™6/TR vector Mammalian Expression Vector (Invitrogen™)

The pcDNA™6⁄TR vector is part of the T-REx™ System (catalog K102001), which yields higher levels of induced expression than any other regulated mammalian expression system. It utilizes the complete CMV promoter and adds control elements from the bacterial tetracycline resistance operon to effectively repress and derepress transcription from one of the strongest mammalian promoter sequences known (1,2).

The regulatory vector, pcDNA™6⁄TR, is provided for high-level expression of the tetracycline repressor (TR) protein. This vector expresses the Blasticidin resistance gene for rapid selection of mammalian cell lines that stably express the TR protein.

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 reverse-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

TA Cloning™ Kit, with pCR™2.1 Vector and One Shot™ TOP10 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.

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
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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.

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
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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

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.

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.

PichiaPink™ Vector Kit (Invitrogen™)

The PichiaPink™ Vector Kit contains the pPINK-HC and pPINK-LC vectors for use with the PichiaPink™ Yeast Expression System. The pPINK vectors are built around the ADE2 gene for complementing the ade2 deletion in the PichiaPink™ strains. You can order the PichiaPink™ Vector Kit to refill your PichiaPink™ Yeast Expression System Kits.

The PichiaPink™ Vector Kit comes with:
•     pPINK-LC vector (low-copy number) (20 µg)
•     pPINK-HC vector (high-copy number) (20 µg)(1)
•     5’AOX1 sequencing primer (20 µg)
•     3’CYC3 sequencing primer (20 µg)
•     One Shot® TOP10 Electrocomp™ E. coli (Cat. No. C404052)

Two Vector Options
The PichiaPink™ Vector Kit comes with two vectors: pPINK-LC (low-copy number) and pPINK-HC (high-copy number). You can choose the vector that works better at producing your protein. Both vectors use the methanol-induced AOX1 promoter to express your protein.

Both pPINK-LC and pPINK-HC vectors are compatible with the PichiaPink™ Secretion Signal Set. Using a 3-way ligation, you can add one of 8 secretion sequences to your gene. The pPINK vectors express proteins intracellularly by default.

Why Choose the PichiaPink™ Yeast Expression System?
The PichiaPink™ Yeast Expression System is based on the yeast Pichia pastoris. Advantages of Pichia pastoris include rapid growth, well-defined genetic background, simple media formulation, and easy handling. For over 30 years, Pichia pastoris has been used by labs around the world for producing hundreds of different proteins from many species including human (2,3). The PichiaPink™ Yeast Expression System allows convenient and cost-effective protein production from small to large scales.

For information on obtaining a commercial-use license for the PichiaPink™Yeast Expression System, please inquire at outlicensing@lifetech.com.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

Related Links
Download the pPINK-LC plasmid map (PDF).
Download the pPINK-HC plasmid map (PDF).
Learn more about the PichiaPink™ Yeast Expression System.
Learn more about our other protein expression systems.

References

1. Du, et al. (2012) Bioengineered Bugs 3:1, 32-37.
2. Cereghino JL, Cregg JM. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev. 2000 Jan;24(1):45-66. [PubMed]
3. Cereghino GP, Cereghino JL, Ilgen C, Cregg JM. Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris. Curr Opin Biotechnol. 2002 Aug;13(4):329-32. [PubMed]

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

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

The Thermo Scientific pTK-Gaussia Luc vector is a derivative of the pMCS-Gaussia Luc vector containing the Gaussia luciferase gene under the control of the Herpes Simplex Virus (HSV) thymidine kinase (TK) promoter. This constitutive expression vector can be used as a normalization control to account for experimental variation in combination with other reporters.

Features of the pCMV-Gaussia Luc vector:

• Weak (TK) constitutive promoters for co-transfection and normalization
• Naturally-secreting Gaussia 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 Gaussia Luc Vectors contain a gene cloned from the copepod, Gaussia princeps. The gene encodes a naturally-secreted bioluminescent Gaussia luciferase (approx. 20kDa), which enables measurement of the reporter activity in media (for real-time assays) and in cell lysates.

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

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pREP4 Mammalian Expression Vector (Invitrogen™)

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

Champion™ pET302/NT-His and pET303/CT-His Vector Kit (Invitrogen™)

The Champion™ pET302/NT-His and pET303/CT-His Vector Kit is designed for cloning a gene of interest via restriction enzyme(s) and ligase (REaL) and subsequent high-level expression from 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 (pET302/NT-His and pET303/CT-His, respectively) for convenient purification and detection (Figure 1). Expression levels obtained from these vectors may be higher than those obtained from another suppliers pET vector (Figure 2). To maximize expression, use with MagicMedia™ E. coli Expression Medium.


Contents and Storage:

The Champion™ pET302/NT-His and pET303/CT-His Vector Kit includes 6 µg each of pET302/NT-His and pET303/CT-His and 10 µg of a control vector. Store at -20“C. All components are guaranteed stable for 6 months when properly stored.

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

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

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

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

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

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

Gateway™ pDONR™/Zeo Vector (Invitrogen™)

Gateway® Technology enables rapid cloning of one or more genes into virtually any protein expression system. Once you have an entry clone, you can recombine your gene of interest into a variety of expression vectors adapted for use in Gateway® Technology. The PCR product is directionally cloned with efficiencies typically >99%. The pDONR™/Zeo vector haa a pUC origin for high plasmid yields and universal M13 sequencing sites for ease of use.

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.

Gateway™ pENTR™ 11 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. New: pENTR™ Dual Selection vectors!

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