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GeneCorner plasmid details

Last data update: 04 December 2020 04:17 CET

Plasmid name: pBRphoA (LMBP 2954)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: pbrphoa.gb (View with Genome Compiler)
pbrphoa.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Escherichia coli alkaline phosphatase A gene (phoA); signal sequence
Promoter: Escherichia coli hybrid tryptophan/lacUV5 promoter (tac)
Ribosome
binding site:
Ribosome binding site (RBS) of the Escherichia coli alkaline phosphatase A gene (phoA)
Terminator: -
Selection marker: Ampicillin (amp)
Tetracycline (tet)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Parental clone: pBR322; pUC19PS1
Further information: This plasmid was constructed by inserting the EcoRI-HindIII fragment from pUC19PS1, containing the Escherichia coli hybrid tryptophan/lacUV5 promoter (tac), the ribosome binding site and the signal sequence of the E. coli alkaline phosphatase A gene (phoA), between the EcoRI and HindIII sites of pBR322.
This plasmid can be used as a secretion vector for bacteria by inserting a gene of interest in phase with the phoA signal sequence.
The presence of the lacI(q) repressor gene is necessary to control the activity of the tac promoter.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727562.1.
EMBL Accession number: LT727562.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 17/12/1993
Sequence detail:
Nucleotide sequence at the end of the phoA signal sequence:

                 *
5' GTG.ACA.AAA.GCG.GTA.CCC.GGG.GAT.CCT.CTA.GAG.TCG.ACC 3'
                 KpnI  SmaI          XbaI    SalI 

*: End of the signal sequence of phoA.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: PstI, SalI and XbaI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr K. De Sutter(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Flamez et al., J. Biotechnol. 42 (1995), 133-143 [PMID: 7576533]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml) + tetracycline (10 μg/ml)
Cultivation temperature: 28°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pBRphoA (LMBP 2954) is available at BCCM/GeneCorner. This plasmid was deposited by Dr K. De Sutter and was published in Flamez et al., 1995.

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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