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GeneCorner plasmid details

Last data update: 20 October 2020 04:14 CEST

Plasmid name: pBM6DraA8 (LMBP 2208)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2208.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse interleukin 6 cDNA (Il6, Il-6, GeneID 16193)
Promoter: Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Phage SP6 promoter
Phage T7 gene 10 promoter (T7g10)
Rous sarcoma virus long terminal repeat (RSV-LTR)
Ribosome
binding site:
-
Terminator: Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Neomycin (neo; G418; kanamycin (kan))
Suppressor tRNA gene (supF; requires a p3 containing host)
Replicon: Bacteriophage M13 origin
Escherichia coli plasmid pMB1 origin
Polyoma virus origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli; use strains with the p3 helper plasmid (kan resistant, amber codon in amp and tet)
Mammalian cells; SV40 permissive cells
Parental clone: pBM5Neo; pB6Delta2
Further information: This plasmid contains the mouse interleukin 6 (mIL6) coding sequence in the antisense orientation relative to the human cytomegalovirus immediate early promoter (CMV-IE) and enhancer.
The plasmid contains the neomycin resistance gene (confers kanamycin resistance in bacteria), which is expressed under the control of the Rous sarcoma virus long terminal repeat (RSV-LTR) as the promoter.
The supF tRNA suppressor gene is used for selection in E. coli MC1061[p3]. It is recommended to select on both ampicillin and tetracycline containing media.
The sequence of the RSV-LTR region corresponds with the Genbank accession number J02025 (version GI:210255).
The nucleotide sequence of the mIL6 cDNA corresponds with the Genbank accession number X06203.1.
The nucleotide sequence at the 5' end of the mIL6 insert into the EcoRV cleaved pBM5Neo vector is not exactly known.
EMBL Accession number: X06203.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 13/06/1991
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 Dr J. Van Snick(1).
(1) Ludwig Institute, Brussels, Belgium
Plasmid reference: -
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061[p3]
Host reference: -
Related host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Invitrogen Instruction Manual
Helper plasmid: p3
Cultivation medium: LB-Lennox + ampicillin (50 μg/ml) + kanamycin (50 μg/ml) + tetracycline (10 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: Statement from Invitrogen (08/11/2002): The spontaneous reversion rate for the amber mutation in the ampicillin marker is 5%, while the reversion rate for the amber mutation in the tetracycline marker is 1%. Since revertant cells (without the SupF plasmid) will grow much faster than cells with the supF plasmid, reversion rates must be minimized using both tetracycline (10 μg/ml) and ampicillin (50 μg/ml) together for selection of E. coli (p3) cells transformed with the SupF plasmid.
Other culture collection numbers: -

Refer in your Materials and Methods:

pBM6DraA8 (LMBP 2208) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. Van Snick.

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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