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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMcHIs (LMBP 2586)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2586.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Escherichia coli outer membrane protein II gene (ompA); signal sequence
Human immunoglobulin γ3 cDNA (hIG3); heavy chain hinge region
Promoter: -
Ribosome
binding site:
-
Terminator: Phage fd terminator
Selection marker: Chloramphenicol (cam)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Parental clone: pMcHB2Mm4s; pMacBLAL2HI
Further information: The plasmid was constructed by substituting the HindIII (blunted with Klenow) - SacI fragment of pMcHB2Mm4s, encoding mutated mature human β2 microglobulin, by the 365 bp DraI-SacI fragment of pMacBLAL2HI,
encoding the human γ3 hinge region.
The humanγ3 hinge region is fused in phase to the signal sequence of the E. coli outer membrane protein II (sompA). The translation of this fusion protein stops two amino acids behind the fourth hinge region (H4),
caused by the presence of a termination codon in the CH2 domain.
The sequence of the inserted hIG3 fragment was taken from 'GENBANK Release 55; Locus: HUMIGHAF' and contains some non-identified nucleotides (indicated by N).
The sompA-Hi fusion is removable with XbaI and AvaI, BamHI, EcoRI or SmaI.
This phasmid provides resistance to chloramphenicol but is sensitive to ampicillin.
After mutagenesis, use mutS strains for primary transformation (e.g. sup(-) strain WK6mutS, sup(+) strain BMH71-18mutS); for segregation of possible mutants: sup(-) strains (e.g. WK6).
EMBL Accession number: J00231, view at EMBL, GenBank, DDBJ
Latest sequence update: 23/01/1997
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) (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 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 + chloramphenicol (25 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
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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