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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pJBLT10lacZ (LMBP 3589)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p3589.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Escherichia coli lac Z gene (lacZ)
Broad-host-range plasmid RK2 trans-acting replication initiator protein DNA (trfA)
Promoter: Phage λ major leftward promoter (λ PL)
Phage T7 gene 10 promoter (T7g10)
Phage T3 promoter
Transposon Tn5 neomycin resistance gene promoter (neo)
Ribosome
binding site:
Ribosome binding site (RBS) of the phage T7 gene 10 (T7g10)
Terminator: Phage fd terminator
Phage T7 gene 10 terminator (T7g10)
Selection marker: Kanamycin (kan)
Ampicillin (amp)
Replicon: Broad-host-range plasmid RK2 vegetative replication origin
Broad-host-range plasmid RK2/RP4 origin of transfer (oriT)
Host range: Escherichia coli
Gram-negative bacterial strains
Parental clone: pJB3kan1; pBBRLT10lacZ
Further information: This IncP plasmid is constructed by inserting the large EagI/ScaI fragment (5584 bp) from pBBRLT10lacZ, containing the lacZ expression cassette, between the BglII and StuI sites of pJB3kan1. The EagI and BglII sites were blunted with Klenow DNA polymerase and the inserted lacZ gene is antisense orientated relative to the ampicillin and kanamycin resistance genes.
The plasmid RK2 trfA gene encodes two polypeptide products P382 and P285 of 382 and 285 amino acids respectively, using a different initiation codon within the same reading frame. The resulting protein is essential for vegetative replication.
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).
EMBL Accession number: -
Latest sequence update: 12/01/2001
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: -
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061(λ)
Host reference: Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329]
Related host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Cultivation medium: LB-Lennox + kanamycin (50 μg/ml)
Cultivation temperature: 28°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pJBLT10lacZ (LMBP 3589) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr E. Remaut .

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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