PB-EF1α-MCS-IRES-GFP PiggyBac cDNA Cloning and Expression Vector

Easily deliver your gene-of-interest using PiggyBac – this vector uses the EF1α promoter to co-expresses your cDNA and a GFP reporter via an IRES element
  • Make transgenic cell lines with a single transfection
  • Integrate multiple PiggyBac Vectors in a single transfection
  • Insert an expression cassette into human, mouse, and rat cells
  • Deliver virtually any-sized DNA insert, from 10 – 100 kb
  • Choose from PiggyBac Vectors that express your gene-of-interest from constitutive or inducible promoters and include a variety of markers

Products

Catalog Number Description Size Price Quantity Add to Cart
PB530A-2 PB-EF1α-MCS-IRES-GFP cDNA cloning and expression vector 10 µg $625
- +

Overview

Overview

Get easy, consistent transgenesis Consistent and easy-to-use, SBI’s PiggyBac Transposon System includes cloning and expression vectors that come with a range of markers as well as both constitutive and inducible promoters. The PB-EF1α-MCS-IRES-GFP PiggyBac cDNA Cloning and Expression Vector (Cat.# PB530A-2) drives co-expression of your gene-of-interest and GFP from the moderate EF1α promoter. Co-expression is mediated by an IRES element upstream of GFP. PB-EF1α-MCS-IRES-GFP PiggyBac cDNA Cloning and Expression VectorWhy use the PiggyBac Transposon System?

Easy, consistent transgenesis with no limits on cargo size—For transgenesis that’s easy, consistent, and not limited by cargo size, SBI’s PiggyBac Transposon System is an excellent choice. The system consists of a PiggyBac Vector and the Super PiggyBac Transposase which recognizes transposon-specific inverted terminal repeats (ITRs) and efficiently integrates the ITRs and intervening DNA into the genome at TTAA sites. The Super PiggyBac Transposase is delivered to the cell via the Super PiggyBac Transposase Expression Vector, which is co-transfected with one or more PiggyBac Vectors.

Footprint-free removal that leaves no PiggyBac sequences behind—In addition to ease-of-use, consistency, and the lack of limits on DNA insert size, what sets this system apart is the ability to reverse the integration reaction in a footprint-free way—with the Excision Only PiggyBac Transposase (Cat.# PB220PA-1), the ITRs and cargo that the Super PiggyBac Transposase integrates into the genome can be removed, leaving behind the original genomic sequence and nothing else.

  • Make transgenic cell lines with a single transfection
  • Integrate multiple PiggyBac Vectors in a single transfection
  • Insert an expression cassette into human, mouse, and rat cells
  • Deliver virtually any-sized DNA insert, from 10 – 100 kb
  • Choose from PiggyBac Vectors that express your gene-of-interest from constitutive or inducible promoters and include a variety of markers
  • Determine the number of integration events with the PiggyBac qPCR Copy Number Kit (# PBC100A-1)

Customer Agreements Academic customers can purchase PiggyBac Transposon System components for internal research purposes for indefinite use, whereas commercial customers must sign a customer agreement for a four-month, limited-use license to evaluate the technology.

For end user license information, see the following:

* SBI is fully licensed to distribute PiggyBac vectors as a partnership with Hera BioLabs, Inc.

How It Works

How It Works

The PiggyBac Transposon System’s Cut-and-Paste Mechanism

The efficient PiggyBac Transposon System uses a cut-and-paste mechanism to transfer DNA from the PiggyBac Vector into the genome. If only temporary genomic integration is desired, the Excision-only PiggyBac Transposase can be transiently expressed for footprint-free removal of the insert, resulting in reconstitution of the original genome sequence.

The PiggyBac Transposon System’s cut-and-paste mechanism

Figure 1. The PiggyBac Transposon System’s cut-and-paste mechanism.

  • The Super PiggyBac Transposase binds to specific inverted terminal repeats (ITRs) in the PiggyBac Cloning and Expression Vector and excises the ITRs and intervening DNA.
  • The Super PiggyBac Transposase inserts the ITR-Expression Cassette-ITR segment into the genome at TTAA sites.
  • The Excision-only Super PiggyBac Transposase can be used to remove the ITR-Expression Cassette-ITR segment from the genome, for footprint-free removal

Supporting Data

Supporting Data

One transfection can integrate one or more genes that can be precisely removed

Efficient transgenesis with the Super PiggyBac Transposase and both single- and dual-promoter PiggyBac Vectors

Figure 2. Efficient transgenesis with the Super PiggyBac Transposase and both single- and dual-promoter PiggyBac Vectors. (Top four panels) Co-transfection with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) and a Dual Promoter PiggyBac Cloning and Expression Vector (Cat.# PB513B-1) into HeLa cells demonstrates the efficient integration delivered by SBI’s PiggyBac Transposon System. After ten days of puromycin selection, only the cells co-transfected with the Super PiggyBac Transposase (+PB, right two panels) show robust growth and GFP fluorescence. (Bottom four panels) Co-transfection with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) and a Single Promoter PiggyBac Cloning and Expression Vector (Cat.# PB531A-2) into HEK293 cells further demonstrates the efficient integration delivered by SBI’s PiggyBac Transposon System. After seven days of growth, the majority of cells that received the Super PiggyBac Transposase Expression Vector (+PB, right two panels) were RFP positive.

Simultaneous integration of multiple PiggyBac Vectors is also highly efficient

Figure 3. Simultaneous integration of multiple PiggyBac Vectors is also highly efficient. METHODS: Three different PiggyBac transposon vectors (Cat.# PB513B-1, Cat.# PB533A-2, and Cat.# PB531A-2) were co-transfected with (left panels) or without (right panels) the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) into Human HT1080 cells. Puromycin and neomycin selection was applied for seven days. The cells that were co-transfected with the Super PiggyBac Transposase Expression Vector were puro and neo resistant, GFP-positive, and RFP-positive. Background GFP-positive cells that are puro resistant stem from random PB513B-1 integrations during the puromycin selection. The non-PiggyBac-mediated integration rate in those cells was extremely low and no RFP-positive cells were identified.

Resources

Citations

  • Uchino, S, et al. (2022) Live imaging of transcription sites using an elongating RNA polymerase II-specific probe. The Journal of cell biology. 1970 Jan 1; 221(2). PM ID: 34854870
  • Teixeira, A, et al. (2022) CelloSelect – A synthetic cellobiose metabolic pathway for selection of stable transgenic CHO cell lines. Metabolic Engineering. 1970 Jan 1; 70:23-30. Link: Metabolic Engineering
  • Rui, Y, et al. (2022) High-throughput and high-content bioassay enables tuning of polyester nanoparticles for cellular uptake, endosomal escape, and systemic in vivo delivery of mRNA. Science advances. 1970 Jan 1; 8(1):eabk2855. PM ID: 34985952
  • Vásquez-Limeta, A, et al. (2022) CPAP insufficiency leads to incomplete centrioles that duplicate but fragment. The Journal of cell biology. 1970 Jan 1; 221(5). PM ID: 35404385
  • Su, CJ, et al. (2022) Ligand-receptor promiscuity enables cellular addressing. Cell systems. 1970 Jan 1;. PM ID: 35421362
  • Klumpe, HE, et al. (2022) The context-dependent, combinatorial logic of BMP signaling. Cell systems. 1970 Jan 1;. PM ID: 35421361
  • Kitano, H, Kawabe, Y & Kamihira, M. (2022) HepG2-Based Designer Cells with Heat-Inducible Enhanced Liver Functions. Cells. 1970 Jan 1; 11(7). PM ID: 35406758
  • Sugiman-Marangos, SN, et al. (2022) Structures of distant diphtheria toxin homologs reveal functional determinants of an evolutionarily conserved toxin scaffold. Communications biology. 1970 Jan 1; 5(1):375. PM ID: 35440624
  • Ma, X, et al. (2022) Validation of reliable safe harbor locus for efficient porcine transgenesis. Functional & integrative genomics. 1970 Jan 1;. PM ID: 35412198
  • Nishimura, K, et al. (2022) Rapid conversion of human induced pluripotent stem cells into dopaminergic neurons by inducible expression of two transcription factors. Stem cells and development. 1970 Jan 1;. PM ID: 35420042
  • Liu, Z, Ramirez, A & Liu, X. (2022) Live Cell Imaging of Spatiotemporal Ca2+ Fluctuation Responses to Anticancer Drugs. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2488:227-236. PM ID: 35347692
  • Dandridge, S. (2022) Honors Thesis: Defining the effect of zinc on the proliferation of MDA-MB-231 cells compared to MCF10A cells. Thesis. 1970 Jan 1;. Link: Thesis
  • Yang, D, et al. (2022) Lineage tracing reveals the phylodynamics, plasticity, and paths of tumor evolution. Cell. 1970 Jan 1; 185(11):1905-1923.e25. PM ID: 35523183
  • Biswas, S, et al. (2022) Long-term hepatitis B virus infection of rhesus macaques requires suppression of host immunity. Nature communications. 1970 Jan 1; 13(1):2995. PM ID: 35637225
  • Breau, KA, et al. (2022) Efficient transgenesis and homology-directed gene targeting in monolayers of primary human small intestinal and colonic epithelial stem cells. Stem cell reports. 1970 Jan 1;. PM ID: 35523179
  • Lensch, S, et al. (2022) Dynamic spreading of chromatin-mediated gene silencing and reactivation between neighboring genes in single cells. eLife. 1970 Jan 1; 11. PM ID: 35678392
  • Gu, J, Sumer, H & Cromer, B. (2022) Efficient Generation of Stable Cell Lines with Inducible Neuronal Transgene Expression Using the piggyBac Transposon System. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2495:49-66. PM ID: 35696027
  • Wang, S, et al. (2021) Budding epithelial morphogenesis driven by cell-matrix versus cell-cell adhesion. Cell. 1970 Jan 1;. PM ID: 34133940
  • Ng, YH, et al. (2021) Efficient generation of dopaminergic induced neuronal cells with midbrain characteristics. Stem cell reports. 1970 Jan 1;. PM ID: 34171286
  • Ukaji, T, et al. (2021) Novel knock-in mouse model for the evaluation of the therapeutic efficacy and toxicity of human podoplanin-targeting agents. Cancer science. 1970 Jan 1; 112(6):2299-2313. PM ID: 33735501
PB-EF1α-MCS-IRES-GFP PiggyBac cDNA Cloning and Expression Vector $625.00

Products

Catalog Number Description Size Price Quantity Add to Cart
PB530A-2 PB-EF1α-MCS-IRES-GFP cDNA cloning and expression vector 10 µg $625
- +

Overview

Overview

Get easy, consistent transgenesis Consistent and easy-to-use, SBI’s PiggyBac Transposon System includes cloning and expression vectors that come with a range of markers as well as both constitutive and inducible promoters. The PB-EF1α-MCS-IRES-GFP PiggyBac cDNA Cloning and Expression Vector (Cat.# PB530A-2) drives co-expression of your gene-of-interest and GFP from the moderate EF1α promoter. Co-expression is mediated by an IRES element upstream of GFP. PB-EF1α-MCS-IRES-GFP PiggyBac cDNA Cloning and Expression VectorWhy use the PiggyBac Transposon System?

Easy, consistent transgenesis with no limits on cargo size—For transgenesis that’s easy, consistent, and not limited by cargo size, SBI’s PiggyBac Transposon System is an excellent choice. The system consists of a PiggyBac Vector and the Super PiggyBac Transposase which recognizes transposon-specific inverted terminal repeats (ITRs) and efficiently integrates the ITRs and intervening DNA into the genome at TTAA sites. The Super PiggyBac Transposase is delivered to the cell via the Super PiggyBac Transposase Expression Vector, which is co-transfected with one or more PiggyBac Vectors.

Footprint-free removal that leaves no PiggyBac sequences behind—In addition to ease-of-use, consistency, and the lack of limits on DNA insert size, what sets this system apart is the ability to reverse the integration reaction in a footprint-free way—with the Excision Only PiggyBac Transposase (Cat.# PB220PA-1), the ITRs and cargo that the Super PiggyBac Transposase integrates into the genome can be removed, leaving behind the original genomic sequence and nothing else.

  • Make transgenic cell lines with a single transfection
  • Integrate multiple PiggyBac Vectors in a single transfection
  • Insert an expression cassette into human, mouse, and rat cells
  • Deliver virtually any-sized DNA insert, from 10 – 100 kb
  • Choose from PiggyBac Vectors that express your gene-of-interest from constitutive or inducible promoters and include a variety of markers
  • Determine the number of integration events with the PiggyBac qPCR Copy Number Kit (# PBC100A-1)

Customer Agreements Academic customers can purchase PiggyBac Transposon System components for internal research purposes for indefinite use, whereas commercial customers must sign a customer agreement for a four-month, limited-use license to evaluate the technology.

For end user license information, see the following:

* SBI is fully licensed to distribute PiggyBac vectors as a partnership with Hera BioLabs, Inc.

How It Works

How It Works

The PiggyBac Transposon System’s Cut-and-Paste Mechanism

The efficient PiggyBac Transposon System uses a cut-and-paste mechanism to transfer DNA from the PiggyBac Vector into the genome. If only temporary genomic integration is desired, the Excision-only PiggyBac Transposase can be transiently expressed for footprint-free removal of the insert, resulting in reconstitution of the original genome sequence.

The PiggyBac Transposon System’s cut-and-paste mechanism

Figure 1. The PiggyBac Transposon System’s cut-and-paste mechanism.

  • The Super PiggyBac Transposase binds to specific inverted terminal repeats (ITRs) in the PiggyBac Cloning and Expression Vector and excises the ITRs and intervening DNA.
  • The Super PiggyBac Transposase inserts the ITR-Expression Cassette-ITR segment into the genome at TTAA sites.
  • The Excision-only Super PiggyBac Transposase can be used to remove the ITR-Expression Cassette-ITR segment from the genome, for footprint-free removal

Supporting Data

Supporting Data

One transfection can integrate one or more genes that can be precisely removed

Efficient transgenesis with the Super PiggyBac Transposase and both single- and dual-promoter PiggyBac Vectors

Figure 2. Efficient transgenesis with the Super PiggyBac Transposase and both single- and dual-promoter PiggyBac Vectors. (Top four panels) Co-transfection with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) and a Dual Promoter PiggyBac Cloning and Expression Vector (Cat.# PB513B-1) into HeLa cells demonstrates the efficient integration delivered by SBI’s PiggyBac Transposon System. After ten days of puromycin selection, only the cells co-transfected with the Super PiggyBac Transposase (+PB, right two panels) show robust growth and GFP fluorescence. (Bottom four panels) Co-transfection with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) and a Single Promoter PiggyBac Cloning and Expression Vector (Cat.# PB531A-2) into HEK293 cells further demonstrates the efficient integration delivered by SBI’s PiggyBac Transposon System. After seven days of growth, the majority of cells that received the Super PiggyBac Transposase Expression Vector (+PB, right two panels) were RFP positive.

Simultaneous integration of multiple PiggyBac Vectors is also highly efficient

Figure 3. Simultaneous integration of multiple PiggyBac Vectors is also highly efficient. METHODS: Three different PiggyBac transposon vectors (Cat.# PB513B-1, Cat.# PB533A-2, and Cat.# PB531A-2) were co-transfected with (left panels) or without (right panels) the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) into Human HT1080 cells. Puromycin and neomycin selection was applied for seven days. The cells that were co-transfected with the Super PiggyBac Transposase Expression Vector were puro and neo resistant, GFP-positive, and RFP-positive. Background GFP-positive cells that are puro resistant stem from random PB513B-1 integrations during the puromycin selection. The non-PiggyBac-mediated integration rate in those cells was extremely low and no RFP-positive cells were identified.

Citations

  • Uchino, S, et al. (2022) Live imaging of transcription sites using an elongating RNA polymerase II-specific probe. The Journal of cell biology. 1970 Jan 1; 221(2). PM ID: 34854870
  • Teixeira, A, et al. (2022) CelloSelect – A synthetic cellobiose metabolic pathway for selection of stable transgenic CHO cell lines. Metabolic Engineering. 1970 Jan 1; 70:23-30. Link: Metabolic Engineering
  • Rui, Y, et al. (2022) High-throughput and high-content bioassay enables tuning of polyester nanoparticles for cellular uptake, endosomal escape, and systemic in vivo delivery of mRNA. Science advances. 1970 Jan 1; 8(1):eabk2855. PM ID: 34985952
  • Vásquez-Limeta, A, et al. (2022) CPAP insufficiency leads to incomplete centrioles that duplicate but fragment. The Journal of cell biology. 1970 Jan 1; 221(5). PM ID: 35404385
  • Su, CJ, et al. (2022) Ligand-receptor promiscuity enables cellular addressing. Cell systems. 1970 Jan 1;. PM ID: 35421362
  • Klumpe, HE, et al. (2022) The context-dependent, combinatorial logic of BMP signaling. Cell systems. 1970 Jan 1;. PM ID: 35421361
  • Kitano, H, Kawabe, Y & Kamihira, M. (2022) HepG2-Based Designer Cells with Heat-Inducible Enhanced Liver Functions. Cells. 1970 Jan 1; 11(7). PM ID: 35406758
  • Sugiman-Marangos, SN, et al. (2022) Structures of distant diphtheria toxin homologs reveal functional determinants of an evolutionarily conserved toxin scaffold. Communications biology. 1970 Jan 1; 5(1):375. PM ID: 35440624
  • Ma, X, et al. (2022) Validation of reliable safe harbor locus for efficient porcine transgenesis. Functional & integrative genomics. 1970 Jan 1;. PM ID: 35412198
  • Nishimura, K, et al. (2022) Rapid conversion of human induced pluripotent stem cells into dopaminergic neurons by inducible expression of two transcription factors. Stem cells and development. 1970 Jan 1;. PM ID: 35420042
  • Liu, Z, Ramirez, A & Liu, X. (2022) Live Cell Imaging of Spatiotemporal Ca2+ Fluctuation Responses to Anticancer Drugs. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2488:227-236. PM ID: 35347692
  • Dandridge, S. (2022) Honors Thesis: Defining the effect of zinc on the proliferation of MDA-MB-231 cells compared to MCF10A cells. Thesis. 1970 Jan 1;. Link: Thesis
  • Yang, D, et al. (2022) Lineage tracing reveals the phylodynamics, plasticity, and paths of tumor evolution. Cell. 1970 Jan 1; 185(11):1905-1923.e25. PM ID: 35523183
  • Biswas, S, et al. (2022) Long-term hepatitis B virus infection of rhesus macaques requires suppression of host immunity. Nature communications. 1970 Jan 1; 13(1):2995. PM ID: 35637225
  • Breau, KA, et al. (2022) Efficient transgenesis and homology-directed gene targeting in monolayers of primary human small intestinal and colonic epithelial stem cells. Stem cell reports. 1970 Jan 1;. PM ID: 35523179
  • Lensch, S, et al. (2022) Dynamic spreading of chromatin-mediated gene silencing and reactivation between neighboring genes in single cells. eLife. 1970 Jan 1; 11. PM ID: 35678392
  • Gu, J, Sumer, H & Cromer, B. (2022) Efficient Generation of Stable Cell Lines with Inducible Neuronal Transgene Expression Using the piggyBac Transposon System. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2495:49-66. PM ID: 35696027
  • Wang, S, et al. (2021) Budding epithelial morphogenesis driven by cell-matrix versus cell-cell adhesion. Cell. 1970 Jan 1;. PM ID: 34133940
  • Ng, YH, et al. (2021) Efficient generation of dopaminergic induced neuronal cells with midbrain characteristics. Stem cell reports. 1970 Jan 1;. PM ID: 34171286
  • Ukaji, T, et al. (2021) Novel knock-in mouse model for the evaluation of the therapeutic efficacy and toxicity of human podoplanin-targeting agents. Cancer science. 1970 Jan 1; 112(6):2299-2313. PM ID: 33735501