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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMc514igam2 (LMBP 3239)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p3239.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Neisseria gonorrhoeae NCTC8375 gene encoding IgA1 protease (iga); mutated sequence
Promoter: Phage λ major leftward promoter (λ PL)
Neisseria gonorrhoeae NCTC8375 gene encoding IgA1 protease promoter (iga)
Ribosome
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: pMc514iga
Further information: This plasmid was derived from pMc514iga by site-specific mutagenesis according to Nakamaye and Eckstein, Nucleic Acids Res. 14 (1986), 9679-9698. As a result, a ScaI site was created at the autoproteolytic site a (the boundary between the protease domain (IgAp) and the IgAαβ domain (IgAAB) of the IgA1 protease gene.
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 may be different 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 may be different from the EMBL-obtained sequence, there is uncertainty as to the cutting frequency of the restriction enzymes indicated.
Other name of the plasmid is pMc514IgA(ScaI).
EMBL Accession number: -
Latest sequence update: 16/06/1995
Sequence detail:
Nucleotide sequence of the mutated IgA1 protease gene.


                        Val Lys Pro Ala Pro|Ser Pro Ala
Wild type seq.: 5' ... .GTC.AAG.CCG.GCC.CCG|TCT.CCT.GCC. ... 3'
                                          @|*


                        Val Lys Pro Ala Ser|Thr Pro Ala
Mutated seq.:   5' ... .GTC.AAG.CCG.GCC.AGT|ACT.CCT.GCC. ... 3'
                                        ^^^|^
                                          @|*
                                        -------
                                        ScaI


@: End of the IgA protease domain (IgAp).
*: Start of the IgAAB domain.
|: Autoproteolytic site a.
^: Mutated nucleotide.
Punctuation indicates reading frame.
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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