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GeneCorner plasmid details

Last data update: 05 December 2020 04:15 CET

Plasmid name: p9HP1B5B12 (LMBP 2794)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2794.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Escherichia coli lac Z gene (lacZ); fragment
Mouse interleukin 6 cDNA (Il6, Il-6, GeneID 16193); fragment of the mature sequence
Promoter: Escherichia coli lac operon promoter
Ribosome
binding site:
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ)
Terminator: -
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Parental clone: pUC9
Further information: The plasmid was constructed by inserting mouse IL6 cDNA into the EcoRI site of pUC9. The 5' end of the mouse IL6 cDNA was partly digested with exonuclease III to remove the 5' untranslated region and the signal sequence.
p9HP1B5B12 is an expression vector for recombinant mouse IL6, a fusion protein consisting of the 8 N-terminal amino acids of bacterial β-galactosidase (excl. Met) fused to amino acid 12 of mature mouse IL6 .
Digestion with EcoRI and AflIII confirmed respectively the length of the plasmid and the orientation of the mouse IL6 gene.
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 at the 3' end of the mIL6 insert is not exactly known. However, digestion with BbsI (BbvII) and SspI confirmed that these sites are still present in the 3' part.
EMBL Accession number: -
Latest sequence update: 03/08/1994
Sequence detail:
Nucleotide sequence at the 5' end of the recombinant mouse IL6 gene.


     1                           8 | 12
Met Thr Met Ile Thr Pro Ser Leu Ala|Thr Glu Asp Thr
ATG.ACC.ATG.ATT.ACG.CCA.AGC.TTG.GCT|ACA.GAG.GAT.ACC
@                                 *|^


@: Initiation codon of the lacZ gene.
*: End of the N-terminal part of the lacZ gene, corresponding
   with amino acid 8 (excl. Met).
^: Start of the mouse IL6 cDNA, corresponding with amino acid 12
   of the mature sequence.

Punctuation indicates reading frame.
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: Simpson et al., Biochem. Biophys. Res. Commun. 157 (1988), 364-372 [PMID: 3264160]
Related plasmid reference: Van Snick et al., Eur. J. Immunol. 18 (1988), 193-197 [PMID: 2965020]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli MC101
Host reference: -
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

p9HP1B5B12 (LMBP 2794) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. Van Snick and was published in Simpson et al., 1988.

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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