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GeneCorner plasmid details

Last data update: 28 October 2020 04:17 CET

Plasmid name: pCAGGS-E-hTRIF (LMBP 5226)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p5226.gb (View with Genome Compiler)
p5226.txt
p5226.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human toll-like receptor (TLR) adapter molecule 1 (TICAM1, hTRIF)
E-tag; N-terminal
Promoter: Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pCAGGS-E-tag
Further information: The plasmid was constructed by inserting the hTRIF coding sequence between the NotI and filled-in EcoRI sites of the pCAGGS-E-tag vector, in frame with the N-terminal E-tag.
pCAGGS-E-hTRIF is useful for highly efficient expression of hTRIF under the control of the AG promoter and the hCMV-IE enhancer in various mammalian cells.
TRIF is an adapter molecule containing a toll/IL1 receptor domain that preferentially activates the hIFNB promoter in the toll-like receptor signaling and is functional at TLR3 and TLR4.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726978.1.
The nucleotide sequence of the hTRIF cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AB093555.1.
Other name of the plasmid is pCAGGS-EhTRIF.
EMBL Accession number: AB093555.1, view at EMBL, GenBank, DDBJ
LT726978.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 29/06/2006
Sequence detail:
Nucleotide sequence at the borders of the fusion gene:

                        ---------------------- E-tag ----------------------
5' ... TTGAATTCCACC.ATG.GGT.GCG.CCG.GTG.CCG.TAT.CCG.GAC.CCG.CTG.GAA.CCG.CGT.GCG.GCC.GCT. 
         EcoRI      Met Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu Glu Pro Arg Ala Ala Ala
                    ***                                                     NotI
                 NcoI


------------ hTRIF ----------------->
 2           5            710     712                                        --> pCAGGS 
GCC.TGC.ACA.GGC...ACG.CAG.GAG.GCA.GAA.TGA.CCCGGGACATTAAACGCGTGGGTACCGGATCCGGCAATTCACT ... 3'
Ala Cys Thr Gly   Thr Gln Glu Ala Glu +++                           BamHI    
                                                                        
     
***: start codon.
+++: termination codon.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, NotI and XbaI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by E. Vercammen(1) (2) and Prof. Dr R. Beyaert(1) (2).
It was constructed by A. Van den Broeke(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Maelfait et al., J. Exp. Med. 205 (2008), 1967-1973 [PMID: 18725521] [DOI: 10.1084/jem.20071632]
Related plasmid reference: Yamomoto et al., J. Immunol. 169 (2002), 6668-6672 [PMID: 12471095]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061
Host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Related host reference: Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pCAGGS-E-hTRIF (LMBP 5226) is available at BCCM/GeneCorner. This plasmid was deposited by E. Vercammen and Prof. Dr R. Beyaert and was published in Maelfait et al., 2008.

Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.

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