Sort By

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.

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.

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.

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

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

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.

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.

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.

T-REx™ Complete Kit, with pcDNA™4/TO/myc-His© Vector (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.

Vivid Colors™ pcDNA™6.2/N-YFP-DEST Vector (Invitrogen™)

The Vivid Colors™ Fluorescent Protein Gateway® Destination Vectors allow you to quickly and easily fuse a protein of interest to the widely used and well-characterized fluorescent proteins from the jellyfish Aequorea victoria (1,2) using the pcDNA™ 6.2 Gateway™ mammalian expression vector. These powerful Gateway® Technology vectors contain the next-generation EGFP, Emerald Green Fluorescent Protein (EmGFP), or the popular Yellow Fluorescent Protein (YFP) for simple, non-invasive detection of recombinant protein. Both fluorescent proteins have been humanized for optimal mammalian expression (3). In addition, the Vivid Colors™ pcDNA™ 6.2 Fluorescent Protein Gateway® Vectors offer:

• CMV promoter for high-level expression of the recombinant fluorescent fusion protein
• Options to fuse EmGFP or YFP to the N- and C-terminus of your protein

pYES2 Yeast Expression Vector (Invitrogen™)

The pYES2 vector is designed for native expression of your protein of interest in S. cerevisiae. It contains the URA3 gene for selection in yeast and 2µ origin for high-copy maintenance.

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]

T-REx™ Core Kit, with pcDNA™4/TO/myc-His© Vector (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.

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

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

Features of the pMCS-Renilla Luc vector:

• Intracellular Green Renilla luciferase gene, optimized for high expression in mammalian systems
• Multiple cloning site (MCS) provides versatility for transfer of regulatory elements from one plasmid to another
• Transcription termination site (Ter), Lac operator (Lac O1), and transcriptional pause site (TPS) used to minimize background by reducing transcriptional read-through
• Both puromycin (Pur) and ampicillin (Amp) markers for drug selection in mammalian and bacterial cells, respectively
• High-copy pUC bacterial DNA replication origin

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

These vectors are subject to a limited use label license.

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

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

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

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

Features of the pTK-Green Renilla Luc vector:

• Weak (TK) constitutive promoter for co-transfection and normalization
• Intracellular Green Renilla luciferase gene, optimized for high expression in mammalian systems
• Transcription termination site (Ter), Lac operator (Lac O1), and transcriptional pause site (TPS) used to minimize background by reducing transcriptional read-through
• Both puromycin (Pur) and ampicillin (Amp) markers for drug selection in mammalian and bacterial cells, respectively
• High-copy pUC bacterial DNA replication origin

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

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