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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPLGN2HIL2 (LMBP 1175)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p1175.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human interleukin 2 cDNA (IL2, IL-2, TCGF, GeneID 3558); mature sequence
Ampicillin resistance gene (amp); signal sequence
Phage λ cI temperature-sensitive repressor gene (cIts, cI857)
Promoter: Escherichia coli β-lactamase promoter (amp)
Phage λ major leftward promoter (λ PL)
Phage λ major rightward promoter (λ PR)
Phage λ promoter for repressor maintenance (λ PRM)
Ribosome
binding site:
Ribosome binding site (RBS) of the Escherichia coli ampicillin resistance gene (amp)
Terminator: -
Selection marker: Kanamycin (kan)
Replicon: Broad-host-range plasmid RSF1010 vegetative replication origin; IncQ
Broad-host-range plasmid RSF1010 origin of transfer (oriT)
Host range: Escherichia coli
Gram-negative bacterial strains
Parental clone: pPLGN1; pPLc2820BHIL2
Further information: The plasmid was constructed by cloning the fusion gene (signal sequence AMP-mature human IL2) as an EcoRI-BamHI(filled in with Klenow DNA polymerase) fragment from pPLc2820BHIL2 between the unique EcoRI and PvuII sites of pPLGN1. The BamHI site is restored.
pPLGN2HIL2 is a broad-host-range plasmid, designed for expression and secretion of human IL2.
It can be used in all gram-negative bacterial species that support replication of the natural plasmid RSF1010.
The plasmid carries the cI857 gene encoding a temperature sensitive repressor. Induction of the PL promoter is obtained at 42°C.
Introduce the plasmid by electroporation, or through conjugation, using plasmid RK2/RP4 transfer functions (provided in trans by RK2/RP4, RK2/RP4-derived vectors, other replicons carrying RK2/RP4 transfer functions (e.g. pRK2073) or strains expressing the tra genes of RK2/RP4). Propagation temperatures may not exceed 34°C, preferentially grow at 28°C.
The nucleotide sequence of the mature human IL2 corresponds with the EMBL Nucleotide Sequence Database accession number X01586.1, except for the first codon of the mature sequence (GCA) that is replaced by GCG.
Other name of the plasmid is pBIL1010.
EMBL Accession number: X01586.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 25/03/2011
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 E. Remaut(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: PhD thesis Ronny Leemans
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 + kanamycin (50 μg/ml)
Cultivation temperature: 28°C
Biosafety level: L1
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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