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GeneCorner plasmid details

Last data update: 30 October 2020 04:11 CET

Plasmid name: pBRphoAE6Hf2 (LMBP 2958)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2958.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 F(ab')2 fragment E6F2 (E6Hf2)
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: pBRphoAE6H; pSV25SE6Hf2
Further information: The plasmid was constructed by ligation of three fragments; the large EcoRI-SalI fragment (ori and amp) and the small EcoRI-PvuII fragment (tac promoter and signal sequence of phoA) of pBRphoAE6H, with the PvuII-SalI fragment of pSV25SE6Hf2 (containing the variable domain, constant part 1 and the hinge region of E6H). This plasmid allows the synthesis of a truncated mouse heavy chain with a CH2 of only 8 amino acids .
In this plasmid the E6H gene of pBRphoAE6H was replaced by the E6Hf2 fragment of pSV23SE6Hf2.
The sphoA-E6Hf2 fusion is in-phase.
The presence of the lacI(q) repressor gene is necessary to control the activity of the tac promoter.
The name mentioned in Flamez et al. (1995) is pBRphoAHf2.
EMBL Accession number: -
Latest sequence update: 04/02/1994
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: 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 MC1061F'
Host reference: -
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml) + tetracycline (10 μg/ml)
Cultivation temperature: 28°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pBRphoAE6Hf2 (LMBP 2958) 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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