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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMxCATlinkDD (LMBP 3908)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p3908.gb (View with Genome Compiler)
p3908.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Chloramphenicol acetyltransferase gene (cat)
Human tumor necrosis factor receptor p55 cDNA (TNFR55, TNFR1, TNFRSF1A); C-terminal fragment of the intracellular domain (death domain)
Promoter: Mouse Mx promoter
Escherichia coli lac operon promoter
Phage SP6 promoter
Ribosome
binding site:
-
Terminator: Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Mammalian cells
Parental clone: pMxCAT; pMxCAThTNFDD
Further information: The plasmid was constructed as follows: a C-terminal fragment of the intracellular domain of the hTNFR55 cDNA, called death domain (DD) and starting with codon 326, was recovered from pMxhTNFR55i and introduced into pMxCAT, in frame at the 3' end of the chloramphenicol acetyltransferase coding sequence and preceded by an oligonucleotide coding for a flexible spacer peptide Gly-Gly-Ser. An additional initiation codon was introduced between the spacer and the hTNFR fragment. In this intermediate plasmid pMxCAThTNFDD, an oligonucleotide was inserted, coding for an extra 15 amino acids linker (Gly4Ser)3, located between the Gly-Gly-Ser linker and the cDNA sequence of the hTNFR55 death domain.
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 LT727283.1.
Other name of the plasmid is pMxCATDD.
EMBL Accession number: LT727283.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 15/01/1999
Sequence detail:
Nucleotide sequence around the fusion between the chloramphenicol acetyltransferase gene and the C-terminal part of the intracellular domain (death domain) of the human tumor necrosis factor receptor p55:
         
         cat -->|            
                |                        spacer
5' … GGC.GGG.GCG.GGC.GGA.TCC.GGT.GGA.GGC.GGT.TCA.GGC.GGA.GGT.GGC.TCT.
     Gly Gly Ala Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 

                        |   |--> human TNF receptor p55
                        |   |--> death domain
                        |   |326                
     GGC.GGT.GGC.GGA.TCC.ATG.GAC.CCC.GCG.ACG.CTG.TAC. … 3'
     Gly Gly Gly Gly Ser Met Asp Pro Tyr Thr Leu Tyr
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglI, EcoNI, HindIII, NcoI, ScaI and XmnI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr G. Haegeman(1) and constructed by Dr V. Vandevoorde(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Vandevoorde et al., J. Cell Biol. 137 (1997), 1627-1638 [PMID: 9199176]
Related plasmid reference: Boone et al., FEBS Lett. 441 (1998), 275-280 [PMID: 9883899]
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: -

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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