LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMc514iga (LMBP 3240)

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


Cloned DNA: Neisseria gonorrhoeae NCTC8375 gene encoding IgA1 protease (iga)
Promoter: Phage λ major leftward promoter (λ PL)
Neisseria gonorrhoeae NCTC8375 gene encoding IgA1 protease promoter (iga)
binding site:
Ribosome binding site (RBS) of the Neisseria gonorrhoeae NCTC8375 gene encoding IgA1 protease (igaT)
Terminator: Phage fd terminator
Selection marker: Chloramphenicol (cam)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli; use strains with a cI function, cIts for PL controlled expression
Parental clone: pMc514; pGEM-7zf(+)IgAp(A)
Further information: The plasmid was constructed by insertion of the IgA1 protease gene of Neisseria gonorhoeae NCTC8375 as a 6.3 kb filled-in (with Klenow DNA polymerase) AvaI fragment from pGEM-7zf(+)IgAp(A) into the filled-in (with Klenow DNA polymerase) BamHI and HindIII sites of pMc514.
After mutagenesis, use mutS and λ lysogen strains for primary transformation; for segregation of possible mutants: sup(-) and λ lysogen strains.
To reconstruct the nucleotide sequence of this plasmid, the sequence of the IgA1 protease gene from Neisseria gonorrhoeae MS11 corresponds with the EMBL Nucleotide Sequence Database (Release 36; Accession number X04835). However, the cloned IgA1 protease gene was derived from Neisseria gonorrhoeae NCTC8375 and differs in several nucleotides.
Since the nucleotide sequences of the 5' and 3' flanking regions of the IgA1 protease gene are not known, and since the nucleotide sequence of the cloned IgA1 protease gene differs from the EMBL-obtained sequence, there is uncertainty as to the cutting frequency of the restriction enzymes indicated.
Other name of the plasmid is pMc514IgAp.
EMBL Accession number: -
Latest sequence update: 13/06/1995
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 I. Poelaert(1) (2) and Prof. Dr E. Remaut(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: PhD thesis Irmgard Poelaert (1995)
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061(λ)
Host reference: Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329]
Related host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Cultivation medium: LB-Lennox + chloramphenicol (25 μg/ml)
Cultivation temperature: 28°C
Biosafety level: L1
Cultivation remark: -
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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