Fluorescent-Protein-Vectors

The Vivid Colors™ Fluorescent Protein Vectors are our next-generation cloning and expression tools featuring Aequoreavictoria-derived fluorescent proteins (FP) for simple, non-invasive detection of recombinant proteins. With Gateway® and TOPO® cloning vector options, cloning, expression, and viewing has never been so easy!

  • Be confident using well-published and trusted FPs derived from Aequorea victoria jellyfish
  • See fluorescent reporter proteins in 24 hours
  • Use multiple colors for co-visualization applications
  • Easily clone into our powerful Gateway® and TOPO® vectors

Spectral properties of Vivid Colors™ fluorescent proteins

Expression of the Vivid Colors™ EmGFP and YFP fusion proteins can be detected by fluorescence microscopy or flow cytometry using standard FITC filter sets. For optimal visibility, it is recommended that you use a filter set optimized for detection within the excitation and emission ranges for each fluorescent protein (see table below).

See excitation and emission profiles for all FPs

 Recommended filters
ProteinColorExcitationEmissionMutationsOmegaChroma
EmGFPGreen487509S65T, S72A, N149K, M153T, I167TXF100 
YFPYellow514527S65G, S72A, K79R, T203YXF104241028
CFPCyan452505K26R, Y66W, N146I, M153T, V163A, N164HXF11431044
BFPBlue308-383440-447F64L, Y66H, Y145F, V163A , N198SXF1031021
Cycle 3 GFPGreen395478507F64L, F100S, M154T, V164AXF76 
 

Vivid Colors™ fluorescent protein fusion vectors

These Vivid Colors™ vectors are for generation of N-terminal or C-terminal fluorescent protein fusions to your gene-of-interest (GOI). Emerald Green (EmGFP) and Yellow (YFP) Fluorescent Protein versions are available. To create a Vivid Colors™ fusion with protein, you can employ either Gateway® or TOPO®-cloning technology. Gateway® Destination Vectors require the use of an entry clone, while the TOPO® vectors require only a PCR product of your GOI (TA Cloning®). Regardless of the cloning techniques you select, the Ultimate ORF™ collection is a comprehensive and convenient source of full-length cDNA clones for creating fusion proteins.

  • Co-expression of your protein with a Vivid Colors™ fluorescent protein
    Express your protein in its native-protein configuration AND EmGFP from a single vector. The Vivid Colors™ pcDNA™ 6.2/ EmGFP-Bsd / V5-DEST vector allows you to rapidly identify transfected cells to look for biological activity conferred by your gene. You’ll need to use a Gateway® entry-clone version of your gene to clone into this destination (DEST) vector. The Ultimate ORF™ collection is a comprehensive and convenient source of full-length entry clones for this purpose. If you already have a clone of your protein, you can amplify it by PCR and clone it with TOPO®-technology to prepare the entry-clone.
  • Create your own vectors with other Vivid Colors™ fluorescent proteins
    The pRSET/CFP (cyan) and pRSET/BFP (blue) vectors contain alternative color versions of fluorescent proteins derived from Aequorea victoria. These proteins are commonly used with the YFP and EmGFP for multi-color applications. Configured for bacterial expression and fluorescent protein purification, these vectors can also be used for subcloning to construct customized mammalian expression vectors.
  • Lentiviral (Vivid Color™) emerald GFP control expression vector
    To optimize lentivial packaging and transduction efficiencies and monitor infected cells, you need the pLenti 6.2/EmGFP-GW Expression Control Vector. This vector allows you to generate high-titer Lentiviral preps that express EmGFP and provides for blasticidin selection. Learn more about viral delivery of mammalian expression vectors.
Other fluorescent protein vectors (Cycle 3 Green Fluorescent Protein)

To express your gene with alternative promoters, or with other selectable markers for stable-cell generation, consider the pTracer™ vectors series. These vectors express the Cycle 3 Green Fluorescent Protein (Cycle 3 GFP) which has spectral excitation and emission wavelengths identical to wild-type GFP. Although nearly as bright as the Vivid Colors™ FPs, you’ll need to use the proper filter set to observe the fully-enhanced brightness.

  • pTracer™/Bsd Expression Vectors
    Choose from CMV or EF-1 a promoters for expressing your gene of interest. The pTracer™ EF vector will allow expression of your protein with carboxy-terminal V-5 and 6xHis epitope tags. The pTracer™ CMV vector is for expression of your protein without any epitope tags. These pTracer™ /Bsd Vectors express the blasticidin resistance gene as a fusion product with the Cycle 3 GFP.
  • pTracer™/Zeo Expression Vectors
    Choose from SV40, CMV or EF-1 a promoters for expressing your gene of interest. The pTracer™ EF vector will allow expression of your protein with carboxy-terminal V-5 and 6xHis epitope tags. The pTracer™ SV40 and CMV vectors are for expression of your protein without any epitope tags. These pTracer™ /Bsd Vectors express the zeocin resistance gene as a fusion product with the Cycle 3 GFP.
  • Promoter Characterization
    If you’re trying to characterize an unknown promoter, you’ll need a cloning and reporter system to monitor expression efficiencies from your PCR-generated mutants. The pGlow and pBlue TOPO® TA Expression Kits provide the convenience of TOPO®-cloning of your PCR products, and the option to quantify promoter activity with either Cycle 3 GFP (pGlow) or b - galactosidase (pBlue) expression.
  • GFP (cycle 3) TOPO® TA Expression Kits
    These vectors are for cloning your PCR-amplified gene of interest to generate a fusion protein with Cycle 3 GFP fused on either the N- or C-termini. With TOPO®-cloning, only a 5-minute reaction is required to generate a clone. Following verification of orientation, the fusion protein is ready to express. The vector elements for these TOPO®-cloning vectors include a CMV promoter for high-level expression of your gene of interest and a neomycin resistance marker.
Highly specific anti-GFP antibodies for Vivid Colors™ Fluorescent Proteins

Highly specific antibodies to Aequorea victoria fluorescent proteins are available, including polyclonal anti-GFP (rabbit serum) and anti-GFP (rabbit IgG fraction), for western blot detection. These antibodies have been tested and are shown to react specifically and strongly to both Vivid Colors™ EmGFP and YFP. A number of different Molecular Probes® anti-GFP antibody conjugates is available, offering you a choice of multiple detection technologies you can employ.

Anti-GFP; Rabbit polyclonal antiserum (A6455)
Anti-GFP; Rabbit polyclonal IgG fraction (A11122)
Anti-GFP; ABfinity® recombinant rabbit monoclonal antibody (G10362)

Summary of fluorescent protein vector elements
 Gene of Interest Mammalian
DescriptionReporterExpr. FormatCloningPromoterTagsSelection
Vivid Colors™ pcDNA™ 6.2/C-EmGFP-DESTEmerald GFPC-terminal FP fusionGateway®CMV bsd
Vivid Colors™ pcDNA™ 6.2/N-EmGFP-DESTEmerald GFPN-terminal FP fusionGateway®CMVN-term.V5bsd
Vivid Colors™ pcDNA™ 6.2/C-YFP-DESTYellowC-terminal FP fusionGateway®CMV bsd
Vivid Colors™ pcDNA™ 6.2/N-YFP-DESTYellowN-terminal FP fusionGateway®CMVN-term.V5bsd
Vivid Colors™ pcDNA™ 6.2/ EmGFP-Bsd/V5-DESTEmerald GFPnativeGateway®CMV GFP-bsd fusion
Vivid Colors™ pcDNA™ 6.2/C-EmGFP-GW/TOPO® Mam. Expr. VectorEmerald GFPC-terminal FP fusionTOPO®CMV bsd
Vivid Colors™ pcDNA™ 6.2/N-EmGFP-GW/TOPO® Mam. Expr. VectorEmerald GFPN-terminal FP fusionTOPO®CMVN-term.V5bsd
Vivid Colors™ pcDNA™ 6.2/C-YFP-GW/TOPO® Mam. Expr. VectorYellowC-terminal FP fusionTOPO®CMV bsd
Vivid Colors™ pcDNA™ 6.2/N-YFP-GW/TOPO® Mam. Expr. VectorYellowN-terminal FP fusionTOPO®CMVN-term.V5bsd
NT-GFP Fusion TOPO TA Expression KitCycle 3 GFPN-terminal FP fusionTOPO®CMV G418
CT-GFP Fusion TOPO TA Expression KitCycle 3 GFPC-terminal FP fusionTOPO®CMV G418
pcDNA™ -DEST47 Gateway® VectorCycle 3 GFPC-terminal FP fusionGateway®CMV G418
pcDNA™ -DEST53 Gateway® VectorCycle 3 GFPN-terminal FP fusionGateway®CMV G418
pBlue TOPO® TA Expression Kitbeta-galn/aTOPO®anyn/aneo
pGlow TOPO® TA Expression KitEmerald GFPn/aTOPO®anyn/aneo
pTracer-CMV/BsdCycle 3 GFPnativeR.E.CMV GFP-bsd fusion
pTracer-CMV/Bsd KitCycle 3 GFPnativeR.E.CMV GFP-bsd fusion
pTracer-EF/Bsd A, B, & CCycle 3 GFPnative + tagR.E.EF-1 alphaC-term.V5, 6xHisGFP-bsd fusion
pTracer-EF/Bsd KitCycle 3 GFPnative + tagR.E.EF-1 alphaC-term.V5, 6xHisGFP-bsd fusion
pTracer-CMV2 KitCycle 3 GFPnativeR.E.CMV GFP-zeo fusion
pTracer-CMV2 VectorCycle 3 GFPnativeR.E.CMV GFP-zeo fusion
pTracer-SV40 KitCycle 3 GFPnativeR.E.SV40 GFP-zeo fusion
pTracer-SV40 VectorCycle 3 GFPnativeR.E.SV40 GFP-zeo fusion
pTracer-EF KitCycle 3 GFPnative + tagR.E.EF-1 alphaC-term.V5, 6xHisGFP-zeo fusion
pTracer-EF A, B, & C VectorCycle 3 GFPnative + tagR.E.EF-1 alphaC-term.V5, 6xHisGFP-zeo fusion
pRSET/CFPCyan(n/a: bacterial expression clone)6xHis, Xpress™, EKn/a
pRSET/BFPBlue(n/a: bacterial expression clone)6xHis, Xpress™, EKn/a
pLenti6.2-GW/EmGFP Expression Control VectorEmerald GFP(GFP control only; no cloning site)CMVn/absd