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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPLc2820BA (LMBP 901)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p901.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Ampicillin resistance gene (amp); signal sequence
Promoter: Escherichia coli β-lactamase promoter (amp)
Phage λ major leftward promoter (λ PL)
Ribosome
binding site:
Ribosome binding site (RBS) of the Escherichia coli ampicillin resistance gene (amp)
Terminator: -
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; use strains with a cI function, cIts for PL controlled expression
Parental clone: pPLc2820BS
Further information: This plasmid contains the β-lactamase signal sequence, preceded by its own ribosome binding site, under control of both the constitutive amp promoter and the thermo-inducible λ PL promoter.
pPLc2820BA is useful to study the effects of both temperature and synthesis level on membrane translocation of genes fused to the amp signal protein.
ApaI and HgiJII can be used for the expression of proteins that are correctly fused with the amp signal protein via removal of the 3' sticky ends.
AvaI, HgiCI, NarI, and SmaI can be used for the expression of proteins that are fused close the amp signal protein.
AvaI, HgiCI, and SmaI sites are not unique.
The amp promoter is not controllable.
The HindIII site cannot be used for the expression of fused proteins, because it is located behind an in-phase termination codon.
Other name of the plasmid is pPLc28-20ß279A.
EMBL Accession number: -
Latest sequence update: 08/06/1990
Sequence detail:
Nucleotide sequence of the ampicillin gene with the polylinker:

5' ATG.AGT.ATT.CAA.CAT.TTC.CGT.GTC.GCC.CTT.ATT.CCC.TTT.TTT.GCG.
    *  


GCA.TTT.TGC.CTT.CCT.GTT.TTT.GCG.GGC.CCG.GGC.GCC.CGG.GCC.TAG.
                                 ^           SmaI       @
                              ApaI      NarI AvaI
                              HgiJII    HgiCI
                                  SmaI
                                  AvaI
AGC.CAA.GCT.T 3'
     HindIII


* : Start of the amp gene.
^ : Start of the mature sequence of the amp gene.
@ : Termination codon
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 Prof. Dr E. Remaut(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Leemans et al., Gene 85 (1989), 99-108 [PMID: 2695405]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 K514(λ)
Host reference: Zabeau et al., EMBO J. 1 (1982), 1217-1224 [PMID: 6327257]
Cultivation medium: LB-Lennox + ampicillin (100 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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