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T7 Cell-Free Expression Vectors


Optimized for in vitro transcription and translation of fusion proteins.

pT7CFE1 cloning plasmids for 1-Step Human IVT Expression System

Thermo Scientific T7 Cell-Free Expression Vectors (pT7CFE1) are cloning plasmids optimized to use with the Thermo Scientific 1-Step Human In Vitro Protein Expression System for in vitro translation (IVT) of tagged fusion proteins.

The pT7CFE1 Expression Vectors contain the Encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES) element that is critical for high levels of cap-independent protein expression in the Human In Vitro Translation System. Each vector features a highly-compatible multiple cloning site (MCS) to facilitate easy insertion of protein coding sequences into and between vectors. The pT7CFE1 Vector is available with single or tandem affinity tags at the N- or C- terminus to facilitate protein purification and detection. The pT7CFE Vectors are suitable for insertion of cloned genes, cDNAs, ORFs or PCR products for in vitro transcription and translation. Custom cloning services are also available.

Highlights:

  • Optimized performance – designed to provide the highest yield in the human in vitro translation system
  • Many options – multiple tag and tag-location options available
  • Modular MCS – multiple cloning site is maintained across vector family to facilitate subcloning
  • Cleavable tags – HRV 3C cleavage site available on select vectors

pT7CFE1 Plasmid Features:

  • EMCV IRES at the 5' UTR promotes high-level translation of mRNAs
  • MCS accommodates gene insertion via ten different restriction sites: Msc1, Nde1, BamH1, EcoR1, EcoRV, Pac1, Pst1, Sac1, Sal1, Not1 and Xho1
  • Poly A sequence in the 3' region promotes mRNA stabilization and protection from nucleases
  • T7 terminator ensures synthesis of accurate size mRNA transcripts
  • Plasmid linearization may be accomplished with restriction sites between Poly A sequence and the T7 terminator region

Product Details:

T7 Cell-free Protein Expression Plasmids
Map of the Thermo Scientific T7 Cell-free Expression Vector elements. Data sheets with features and maps of individual plasmids and complete sequence information are available for download (see table).

 

Thermo Scientific pT7CFE1 Vectors for cell-free protein expression and available affinity and cleavage tag options.
Vector N-term
Tag
C-term
Tag
Cleavage
Site
Vector Information
Files
pT7CFE1-NHis 6xHis     Data sheet Sequence
pT7CFE1-CHis   6xHis   Data sheet Sequence
pT7CFE1-NHA HA     Data sheet Sequence
pT7CFE1-CHA   HA   Data sheet Sequence
pT7CFE1-NMyc c-Myc     Data sheet Sequence
pT7CFE1-CMyc   c-Myc   Data sheet Sequence
pT7CFE1-NFtag Ftag     Data sheet Sequence
pT7CFE1-CFtag   Ftag   Data sheet Sequence
pT7CFE1-NHA-CHis HA 6xHis   Data sheet Sequence
pT7CFE1-CGST-HA-His   GST
HA
6xHis
HRV 3C
(C-term)
Data sheet Sequence
pT7CFF1-CGFP-HA-His   GFP
HA
6xHis
HRV 3C
(C-term)
Data sheet Sequence
pT7CFE1-NHis-GST 9xHis
GST
  HRV 3C
(N-term)
Data sheet Sequence
pT7CFE1-NHis-GST-CHA 9xHis
GST
HA HRV 3C
(N-term)
Data sheet Sequence

 

Importance of IRES element vs 5' cap for in vitro protein expression. IRES-mediated protein expression is significantly greater than 5' capped mRNA. tGFP mRNA was transcribed from the pCFE expression plasmids containing an upstream IRES element (+ IRES) or another plasmid without an IRES element. The mRNA generated without an IRES element was either used directly (- IRES, - cap) or chemically modified to have an N-terminal Anti-Reverse Cap Analog (ARCA, + 5' cap). Equal amounts of all the three mRNA's were used in human in vitro translation reactions for 2 hours at 30°C and the relative amount of GFP was determined by fluorescence.

 

More Product Data

 

Product References:

  1. Yadavalli, R., Ledger, C., Sam-Yellowe, TY. (2012). In vitro human cell-free expression system for synthesis of malaria proteins. Parasitol Res. 2012 Jul 11. [Epub ahead of print] DOl I 0.1007 /s00436-0 12-3014-7.

Related Resources:

Getting Started Guide – overview of critical parameters for successful human in vitro protein expression
Review of cell-free in vitro recombinant protein expression
DNASU library of GST-tagged cell-free expression-ready plasmids
Tech Tip #72 – PCR protocol for generating optimized templates for Pierce Human In Vitro Expression Kits

Related Products:

Custom Expression Vector Construction Service
1-Step Human In Vitro Protein Expression Kits (for coupled and mRNA-template reactions)
1-Step Human High-Yield In Vitro Protein Expression Kits
Epitope Tag and Fusion Tag Antibodies


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
88859 pT7CFE1-NHis
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88859 pT7CFE1-NHis Certificate of Analysis for product #88859 pT7CFE1-NHis $57.00

Add to cart
88860 pT7CFE1-CHis
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88860 pT7CFE1-CHis Certificate of Analysis for product #88860 pT7CFE1-CHis $57.00

Add to cart
88861 pT7CFE1-NHA
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88861 pT7CFE1-NHA Certificate of Analysis for product #88861 pT7CFE1-NHA $57.00

Add to cart
88862 pT7CFE1-CHA
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88862 pT7CFE1-CHA Certificate of Analysis for product #88862 pT7CFE1-CHA $57.00

Add to cart
88863 pT7CFE1-NMyc
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88863 pT7CFE1-NMyc Certificate of Analysis for product #88863 pT7CFE1-NMyc $57.00

Add to cart
88864 pT7CFE1-CMyc
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88864 pT7CFE1-CMyc Certificate of Analysis for product #88864 pT7CFE1-CMyc $57.00

Add to cart
88865 pT7CFE1-NFtag
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88865 pT7CFE1-NFtag Certificate of Analysis for product #88865 pT7CFE1-NFtag $57.00

Add to cart
88866 pT7CFE1-CFtag
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88866 pT7CFE1-CFtag Certificate of Analysis for product #88866 pT7CFE1-CFtag $57.00

Add to cart
88867 pT7CFE1-NHA-CHis
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88867 pT7CFE1-NHA-CHis Certificate of Analysis for product #88867 pT7CFE1-NHA-CHis $57.00

Add to cart
88868 pT7CFE1-CGST-HA-His
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88868 pT7CFE1-CGST-HA-His Certificate of Analysis for product #88868 pT7CFE1-CGST-HA-His $57.00

Add to cart
88869 pT7CFE1-CGFP-HA-His
Formulation: 0.5µg/µL DNA in 10mM Tris-HCl, pH 8.5
10µg MSDS for product #88869 pT7CFE1-CGFP-HA-His Certificate of Analysis for product #88869 pT7CFE1-CGFP-HA-His $57.00

Add to cart
88870 pT7CFE1-NHis-GST
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #88870 pT7CFE1-NHis-GST Certificate of Analysis for product #88870 pT7CFE1-NHis-GST $57.00

Add to cart
88871 pT7CFE1-NHis-GST-CHA
Formulation: 0.5μg/μL DNA in 10mM Tris-HCl, pH 8.5

10μg MSDS for product #88871 pT7CFE1-NHis-GST-CHA Certificate of Analysis for product #88871 pT7CFE1-NHis-GST-CHA $57.00

Add to cart

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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