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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMxhTNFR55i (LMBP 2787)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p2787.gb (View with Genome Compiler)
p2787.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: 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: pSP64MxpA; pEThTNFR55i
Further information: This plasmid was constructed by ligating the following three fragments: the XbaI-BglI and the BglI-BamHI fragment from pSP64MxpA, and the XbaI-BamHI fragment from pEThTNFR55i containing the intracellular part of the hTNFR55 cDNA and part of the 3' UTR.
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 LT727545.1.
Other name of the plasmid is pMxHTNFR1I.
EMBL Accession number: LT727545.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 16/11/1994
Sequence detail:
Nucleotide sequence of the mutated hTNFR55 cDNA:

hTNFR55wt:

        transmembrane domain   -->|
                                  |
                   200            |204
5' TTA TCC CTC CTC TTC ATT GGT TTA ATG TAT CGC 3'
   Leu Ser Leu Leu Phe Ile Gly Leu Met Tyr Arg


pMxhTNFR55i:
                                             |
                                             |204
5' TCTAGAAA...TTTAAGAAGGAGATATACC ATG GGT TTA ATG TAT CGC 3'
   XbaI                           Met Gly Leu Met Tyr Arg
                               NcoI
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: EcoNI, HindIII and NcoI. The results of the EcoNI and HindIII digestions do suppose that the region between position 407 (the EcoNI site in the Mx promoter) and 628 (the HindIII site in the hTNFR55i insert) is some nucleotides longer.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr G. Haegeman(1). It was 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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