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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMac254BLAHI (LMBP 2150)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2150.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Ampicillin resistance gene (amp)
Human immunoglobulin γ3 cDNA (hIG3); heavy chain hinge region
Promoter: Escherichia coli kanamycin resistance gene promoter (kan)
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: pMac254mSA; pBR325OMMBfmA
Further information: The plasmid was constructed by introducing the SacI-ApaI (nucleotide position 5044) fragment of pBR325OMMBfmA, containing the hinge region of the human immunoglobulin γ3, into the unique SacI and ApaI sites of pMac254mSA, creating a fusion gene (BlaHi).
The fusion gene is cloned downstream from the constitutive kanamycin phosphoribosyltransferase promoter.
The ampicillin resistance gene is directly fused to the start of the first hinge region H1.
The fusion product still provides resistance to ampicillin.
The translation of the fusion gene terminates 8 amino acids behind the end of the hinge region.
The f1 part of the phasmid contains the f1 origin, but also the sequence around the gene II promoter.
Name mentioned in De Sutter et al. (1992) is pMacBlaHi.
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: De Sutter et al., Mol. Microbiol. 6 (1992), 2201-2208 [PMID: 1406260]
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 (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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