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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMxCAT(stop)hTNFR55i (LMBP 3906)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p3906.gb (View with Genome Compiler)
p3906.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Chloramphenicol acetyltransferase gene (cat)
Human tumor necrosis factor receptor p55 cDNA (TNFR55, TNFR1, TNFRSF1A); intracellular 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; pMxhTNFR55i
Further information: This plasmid was constructed as follows: the intracellular domain of the hTNFR55 cDNA was recovered from pMxhTNFR55i and introduced into pMxCAT, downstream from the 'TAA' termination codon of the chloramphenicol acetyltransferase gene and preceded by an oligonucleotide coding for a flexible spacer peptide Gly-Gly-Ser-Gly.
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 LT727281.1.
EMBL Accession number: LT727281.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 14/01/1999
Sequence detail:
Nucleotide sequence around the fusion between the chloramphenicol acetyltransferase gene and the intracellular domain of the human tumor necrosis factor receptor p55 cDNA:

                     |               |--> human TNF receptor p55
            cat  --> |               |           |--> intracellular domain
                     |    spacer     |201        |204                    
5' … .GGC.GGG.GCG.TAA.GGC.GGA.TCC.GGC.ATG.GGT.TTA.ATG.TAT.CGC.TAC. … 3'
      Gly Gly Ala     Gly Gly Ser Gly Met Gly Leu Met Gly Leu Tyr
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, BglI, EcoNI, HindIII and ScaI.
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]
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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