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Timer fluorescent protein

Photoactivatable/switchable fluorescent protein

Study promoter activity using the Living Colors fluorescent Timer, a fluorescent protein that shifts color from green to red over time—in a matter of hours, depending on the expression system (Terskikh et al. 2000). Shortly after its synthesis, the fluorescent Timer begins emitting green fluorescence, but as time passes, the fluorophore undergoes additional changes that shift its fluorescence to longer wavelengths. When fully matured, the protein is bright red.

Study promoter activity using the Living Colors fluorescent Timer, a fluorescent protein that shifts color from green to red over time—in a matter of hours, depending on the expression system (Terskikh et al. 2000). Shortly after its synthesis, the fluorescent Timer begins emitting green fluorescence, but as time passes, the fluorophore undergoes additional changes that shift its fluorescence to longer wavelengths. When fully matured, the protein is bright red. Detect the Living Colors fluorescent Timer with the DsRed polyclonal antibody.

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Cat. # Product Size Price License Quantity Details
632402 pTimer Vector 20 ug USD $589.00

License Statement

ID Number  
72 Living Colors Fluorescent Protein Products:

Not-For-Profit Entities: Orders may be placed in the normal manner by contacting your local representative or Takara Bio USA, Inc. Customer Service. Any and all uses of this product will be subject to the terms and conditions of the Non-Commercial Use License Agreement (the “Non-Commercial License”), a copy of which can be found below. As a condition of sale of this product to you, and prior to using this product, you must agree to the terms and conditions of the Non-Commercial License. Under the Non-Commercial License, Takara Bio USA, Inc. grants Not-For-Profit Entities a non-exclusive, non-transferable, non-sublicensable and limited license to use this product for internal, non-commercial scientific research use only. Such license specifically excludes the right to sell or otherwise transfer this product, its components or derivatives thereof to third parties. No modifications to the product may be made without express written permission from Takara Bio USA, Inc. Any other use of this product requires a different license from Takara Bio USA, Inc. For license information, please contact a licensing representative by phone at 650.919.7320 or by e-mail at licensing@takarabio.com.

For-Profit Entities wishing to use this product are required to obtain a license from Takara Bio USA, Inc. For license information, please contact a licensing representative by phone at 650.919.7320 or by e-mail at licensing@takarabio.com.

Not-For-Profit Non-Commercial Use License:
A copy of the pTimer Vector product License Agreement can be found by clicking here.
24 The RCFPs (including DsRedExpress, DsRedExpress2, and E2-Crimson) are covered by one or more of the following U.S. Patent Nos. 7,166,444; 7,157,565; 7,217,789; 7,150,979; 8,093,450 and 8,461,769.
*

pTimer is a prokaryotic expression vector that encodes DsRed1-E5, a fluorescent protein that changes color as it ages. Shortly after translation, DsRed1-E5 emits green light; its red fluorophore emerges later, hours after translation. Because of its predictable color shift, DsRed1-E5 acts as a timer, useful for monitoring changes in gene activity in living cells.

DsRed1-E5 is a mutant of the red fluorescent protein, DsRed1. The wild-type protein, DsRed, was originally referred to as drFP583. DsRed1-E5 contains two amino acid substitutions (V105A and S197T), which increase its fluorescence intensity and endow it with a distinct spectral property: As the protein ages, it changes color. When first synthesized, DsRed1-E5 is bright green (excitation & emission maxima = 483 nm & 500 nm). As time passes, the green fluorophore undergoes additional changes that cause its fluorescence to shift to longer wavelengths-when fully matured, the protein is bright red (excitation & emission maxima = 558 nm & 583 nm). In mammalian cells transfected with a Tet-inducible DsRed1-E5 expression vector, the green-to-red transition starts about 3 hours after the protein first becomes fluorescent. In addition to the amino acid replacements, DsRed1-E5's coding sequence contains a series of silent base-pair changes, which correspond to human codon-usage preferences, for high expression in mammalian cells.

pTimer is primarily intended to serve as a convenient source of the DsRed1-E5 cDNA. The DsRed1-E5 coding sequence is flanked by separate and distinct multiple cloning sites at the 5' and 3' ends so that the gene can easily be excised for use in other expression systems. Alternatively, the DsRed1-E5 coding sequence can be amplified by PCR.

The DsRed1-E5 gene was inserted in frame with the lacZ initiation codon from pUC19 so that DsRed1-E5 is expressed from the lac promoter (Plac) in E. coli host cells. A Kozak consensus sequence is located immediately upstream of DsRed1-E5 to enhance translational efficiency should you wish to express the gene in eukaryotic systems. The entire DsRed1-E5 expression cassette in pTimer is supported by a pUC backbone, which contains a high-copy number origin of replication and an ampicillin resistance gene for propagation and selection in E. coli.

When expressed in multicellular organisms, DsRed1-E5's invariable green-to-red shift can be used as a timer to track the on-off phases of gene expression during embryogenesis and cell differentiation.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

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Vector map for pTimer vectors

Vector map for pTimer vectors
Vector map for pTimer vectors. As the Fluorescent Timer matures, it changes color—in a matter of hours, depending on the expression system used.

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Fluorescent timer under the control of the heat shock promoter hsp16-41 in a transgenic C. elegans embryo

Fluorescent timer under the control of the heat shock promoter hsp16-41 in a transgenic C. elegans embryo

Fluorescent Timer under the control of the heat shock promoter hsp16-41 in a transgenic C. elegans embryo. The embryo was heat shocked in a 33°C water bath. Promoter activity was studied during the heat shock recovery period. Green fluorescence was observed in the embryo as early as two hr into the recovery period. By 50 hr after heat shock, promoter activity had ceased, as indicated by the lack of green color.

632403 pTimer-1 Vector 20 ug USD $589.00

License Statement

ID Number  
72 Living Colors Fluorescent Protein Products:

Not-For-Profit Entities: Orders may be placed in the normal manner by contacting your local representative or Takara Bio USA, Inc. Customer Service. Any and all uses of this product will be subject to the terms and conditions of the Non-Commercial Use License Agreement (the “Non-Commercial License”), a copy of which can be found below. As a condition of sale of this product to you, and prior to using this product, you must agree to the terms and conditions of the Non-Commercial License. Under the Non-Commercial License, Takara Bio USA, Inc. grants Not-For-Profit Entities a non-exclusive, non-transferable, non-sublicensable and limited license to use this product for internal, non-commercial scientific research use only. Such license specifically excludes the right to sell or otherwise transfer this product, its components or derivatives thereof to third parties. No modifications to the product may be made without express written permission from Takara Bio USA, Inc. Any other use of this product requires a different license from Takara Bio USA, Inc. For license information, please contact a licensing representative by phone at 650.919.7320 or by e-mail at licensing@takarabio.com.

For-Profit Entities wishing to use this product are required to obtain a license from Takara Bio USA, Inc. For license information, please contact a licensing representative by phone at 650.919.7320 or by e-mail at licensing@takarabio.com.

Not-For-Profit Non-Commercial Use License:
A copy of the pTimer-1 Vector product License Agreement can be found by clicking here.
24 The RCFPs (including DsRedExpress, DsRedExpress2, and E2-Crimson) are covered by one or more of the following U.S. Patent Nos. 7,166,444; 7,157,565; 7,217,789; 7,150,979; 8,093,450 and 8,461,769.
*

pTimer-1 is a promoterless vector that can be used to monitor transcription from different promoters and promoter/enhancer combinations inserted into the MCS located upstream of the DsRed1-E5 coding sequence. Shortly after translation, DsRed1-E5 emits green light; its red fluorophore emerges later, hours after translation. Because of its predictable color shift, DsRed1-E5 acts as a timer, useful for monitoring changes in gene activity in living cells. pTimer-1 can be used to study the on-off activity of any cis-acting regulatory element cloned into the MCS upstream of DsRed1-E5.

DsRed1-E5 is a mutant of the red fluorescent protein, DsRed1. The wild-type protein, DsRed, was originally referred to as drFP583. DsRed1-E5 contains two amino acid substitutions (V105A and S197T), which increase its fluorescence intensity and endow it with a distinct spectral property: As the protein ages, it changes color. When first synthesized, DsRed1-E5 is bright green (excitation & emission maxima = 483 nm & 500 nm). As time passes, the green fluorophore undergoes additional changes that cause its fluorescence to shift to longer wavelengths-when fully matured, the protein is bright red (excitation & emission maxima = 558 nm & 583 nm). In mammalian cells transfected with a Tet-inducible DsRed1-E5 expression vector, the green-to-red transition starts about 3 hours after the protein first becomes fluorescent. In addition to the amino acid replacements, DsRed1-E5's coding sequence contains a series of silent base-pair changes, which correspond to human codon-usage preferences, for high expression in mammalian cells.

To further increase translation efficiency in eukaryotic cells, a sequence upstream of DsRed1-E5 has been converted to a Kozak consensus sequence. SV40 polyadenylation signals downstream of the DsRed1-E5 gene direct proper processing of the 3' end of the DsRed1-E5 mRNA. The vector backbone contains an SV40 origin for replication in mammalian cells expressing the SV40 T antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. A neomycin-resistance cassette (Neor) allows stably transfected eukaryotic cells to be selected using G418. This cassette consists of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene. A bacterial promoter upstream of the cassette expresses kanamycin resistance in E. coli. Without addition of a functional promoter, this vector will not express DsRed1-E5.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

Back

Vector map for pTimer vectors

Vector map for pTimer vectors
Vector map for pTimer vectors. As the Fluorescent Timer matures, it changes color—in a matter of hours, depending on the expression system used.

Back

Fluorescent timer under the control of the heat shock promoter hsp16-41 in a transgenic C. elegans embryo

Fluorescent timer under the control of the heat shock promoter hsp16-41 in a transgenic C. elegans embryo

Fluorescent Timer under the control of the heat shock promoter hsp16-41 in a transgenic C. elegans embryo. The embryo was heat shocked in a 33°C water bath. Promoter activity was studied during the heat shock recovery period. Green fluorescence was observed in the embryo as early as two hr into the recovery period. By 50 hr after heat shock, promoter activity had ceased, as indicated by the lack of green color.

*You must be logged in to a Business Account in order to purchase these products online, since the purchase of these products may be restricted depending on your account type. Researchers at not-for-profit accounts receive a limited use license with their purchase of the product. Researchers at for-profit accounts must obtain a license prior to purchase. For details please contact licensing@takarabio.com.

Selection guides Fluorescent protein quick guide
Over 170 fluorescent protein vectors for $149 each Fluorescent protein plasmids for $149
Selection guides Fluorescent lentiviral particles

More Information

Applications

  • Promoter studies
  • Visualize the location and time frame of promoter activity (e.g., during embryogenesis and cell differentiation)
References

Terskikh, A. et al. "Fluorescent timer": protein that changes color with time. Science 290(5496):1585–1588 (2000).

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


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