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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMxh5HT1AR (LMBP 3028)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p3028.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human serotonin 5-HT1α receptor (h5HT1AR)
Promoter: Mouse Mx promoter
Escherichia coli lac operon promoter
Phage SP6 promoter
Ribosome
binding site:
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ)
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; pSP64H5HT1AR
Further information: pSP64H5HT1AR was opened with TfiI and treated with Klenow DNA polymerase. After BamHI digestion of the blunted fragment, the human 5HT1α receptor fragment was isolated. pSP64MxpA was cleaved with XbaI, filled in with Klenow DNA polymerase, and after BglI digest the fragment containing the mouse Mx promoter was isolated. pSP64MxpA was also digested with the combination BamHI / BglI in order to isolate the fragment containing the SV40 intron and polyadenylation signals. The three isolated fragments were ligated resulting in the plasmid pMxh5HT1AR in which the human serotonin 5HT1α receptor is under control of the mouse Mx promoter and SV40 processing signals. Cell lines transfected with this plasmid allow functional expression of the receptor in L929s(A), after induction of the Mx promoter with 1000U/ml mIFNα or mIFNβ.
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.
Other name of the plasmid is pMx5HT1A.
EMBL Accession number: -
Latest sequence update: 05/09/1994
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed in the research unit of Em. Prof. Dr G. Haegeman(2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
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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