Last data update: 24 November 2020 04:18 CET
Plasmid name: pMc514igam1 (LMBP 3238)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|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 (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|
|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 BsmI site was created at the start codon of the IgA1 protease gene, making the coding sequence accessible for blunt-end ligation.
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(BsmI).
|EMBL Accession number:||-|
|Latest sequence update:||15/06/1995|
Nucleotide sequence of the mutated IgA1 protease gene. Wild type sequence: 5' ... AAACGGTAAAACCTT ATG.AAA.GCC. ... 3' * Mutated sequence: 5' ... AAACGGTAAAAGAAT GCG.AAA.GCC. ... 3' ^^^ ^^ -------- BsmI *: Start codon of the IgA1 protease gene. ^: Mutated nucleotide. Punctuation indicates reading frame.
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|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)|
|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.