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GeneCorner plasmid details

Last data update: 29 October 2020 04:15 CET

Plasmid name: pBRphoAE6Hf1v (LMBP 2955)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2955.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Escherichia coli alkaline phosphatase A gene (phoA); signal sequence
Mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ) cDNA; truncated heavy chain of the Fab fragment E6F1 (E6Hf1)
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)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Parental clone: pBRphoAE6Hv; pSV23E6Hf1; pBR322
Further information: The plasmid was constructed by ligation of three fragments; the large EcoRI-SalI fragment of pBR322 (ori and amp) and the small EcoRI-PvuII fragment (tac promoter and signal sequence of phoA) of pBRphoAE6Hv, with the PvuII-SalI fragment of pSV23E6Hf1 (containing the variable domain and constant domain 1 of E6H). The translation stops at the beginning of the hinge region.
In this plasmid the E6H gene of pBRphoAE6Hv was replaced by the E6Hf1 fragment of pSV23E6Hf1.
The construction of pBRphoAE6Hv (parental clone of pBRphoAE6Hf1v) is similar to that of pBRphoAE6H, except that the KpnI exonuclease blunting did not occur. This results in a not-in-phase sphoA-E6Hf1 fusion and a restored KpnI site. The EcoRI-KpnI promoter-signal sequence cassette is useful for in-phase construction.
The presence of the lacI(q) repressor gene is necessary to control the activity of the tac promoter.
Other name of the plasmid is pBRphoAE6Hf1(KpnI).
EMBL Accession number: -
Latest sequence update: 16/02/1994
Sequence detail:
Nucleotide sequence of the sphoA-E6Hf1 fusion:


      sphoA   -->|    |<-- VH
                 |    |
5' ...ACA AAA GCG GTAC CAG GTT CAG CTG... 3'
                KpnI           PvuII
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 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: -
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)
Cultivation temperature: 28°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pBRphoAE6Hf1v (LMBP 2955) is available at BCCM/GeneCorner. This plasmid was deposited by Dr K. De Sutter .

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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