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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPSA2 (LMBP 2885)

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Price category: Cat. 3 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: -
Sequence
analysis results
Genecorner:

Sanger: 148921.fasta

Sanger: 148922.fasta

NGS: a05-gc-dec2018.fasta

Cloned DNA: Staphylococcus aureus protein A cDNA (spa); fragment
Promoter: Staphylococcus aureus protein A promoter (psa)
Ribosome
binding site:
Ribosome binding site (RBS) of the Staphylococcus aureus protein A (spa) gene
Terminator: -
Selection marker: Ampicillin (amp)
Chloramphenicol (cam)
Replicon: Escherichia coli plasmid pMB1 origin
Staphylococcal plasmid pC194 origin
Host range: Escherichia coli
Staphylococcus aureus
Parental clone: pRIT21
Further information: The plasmid was constructed based on pRIT21 by introducing an SpeI site upstream of the presumed Shine-Dalgarno sequence of the Staphylococcus aureus protein A (spa) gene and creating an AflIII site to introduce an ATG, replacing the wild type TTG of the spa gene.
Genes of interest can be cloned into pPSA2 by opening the plasmid via AflIII/EcoRI restriction digest.
The region upstream of the ampicillin resistance gene and downstream of the pMB1 ori have been sequenced at BCCM/GeneCorner.
EMBL Accession number: -
Latest sequence update: 22/06/2018
Sequence detail:
Sequence at the start of the S. aureus protein A coding sequence:

5' GACTTTACTAGTACATACAGGGGGTATTAAC.ATG.TAA. 3'
           * *  -----------      * *   *
         SpeI       S.D.

*: mutated nucleotide
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglI/HindIII, EcoRI/PvuI, NcoI and SspI.

This plasmid has also been fully sequenced but the NGS sequence data still need to be implemented.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr L. Steidler(1) (2) and 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: Steidler et al., Appl. Environ. Microbiol. 62 (1996), 2356-2359 [PMID: 8779573]
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 + ampicillin (100 μg/ml) + chloramphenicol (34 μg/ml)*
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: *: selection of transformants on chloramphenicol; subsequent cultivation of a single colony in liquid medium with ampicillin.
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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