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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMc59BLAHI (LMBP 2268)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2268.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Ampicillin resistance gene (amp)
Human immunoglobulin γ3 cDNA (hIG3); heavy chain hinge region
Promoter: -
Ribosome
binding site:
-
Terminator: Phage fd terminator
Selection marker: Ampicillin (amp)
Chloramphenicol (cam)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Parental clone: pMc59; pMac254BLAHI
Further information: The plasmid was constructed by ligating the BssHII (nucleotide position 2926; filled in with Klenow DNA polymerase) - AlwNI (blunted with T4 DNA polymerase) fragment of pMac254BLAHI, containing the fusion gene, in clockwise orientation in the unique XbaI site (filled in with Klenow DNA polymerase) of pMc59.
This pMc59-derivative is similar to pMac254BLAHI, but the fusion gene is clockwise orientated and located between the chloramphenicol resistance and the fd terminators.
The phasmid provides resistance to chloramphenicol. Since it provides also resistance to ampicillin, one can conclude that the constitutive promoter of kanamycin phosphoribosyltransferase is still present in a functional way upstream of the fusion gene.
The translation of the fusion gene stops 8 amino acids behind the end of the hinge region of the human IgG3 heavy chain.
pMc59BLAHI contains the replication origin of the single-stranded DNA phage f1, so that it can be rapidly switched between the plasmid and the phage mode (ssDNA) of replication. The latter requires infection with the helper phage M13KO7.
EMBL Accession number: -
Latest sequence update: 22/01/1997
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).
(1) 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) + chloramphenicol (25 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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