Last data update: 28 November 2020 04:24 CET
Plasmid name: pMG7 (LMBP 8458)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Bacillus subtilis amylase gene (amyE); 5' UTR and 3' UTR
Bacillus subtilis glyceraldehyde 3-phosphate dehydrogenase (gapA); 5' UTR containing transcriptional regulator
|Promoter:||Escherichia coli lac operon promoter
Bacillus subtilis central glycolytic genes regulator promoter (cggR)
|Selection marker:||Ampicillin (amp)
|Replicon:||Escherichia coli plasmid pMB1 origin|
|Host range:||Escherichia coli
Bacillus sp.; integrative
|Further information:||The plasmid was constructed by PCR amplifying the B. subtilis cggR promoter as an EcoRI/PstI fragment and the B. subtilis gapA 5' UTR as a PstI/Acc65I fragment, and cloning them jointly into the EcoRI/Acc65I-opened pMG6 vector.
This plasmid is a modular E. coli–Bacillus shuttle vector, applicable for overproduction of proteins integrated in the chromosome of B. subtilis.
Genes can be integrated in another site by replacing the 5' UTR and 3' UTR of amyE as an AatII/XbaI fragment and SalI/NotI fragment respectively.
Furthermore, the modular architecture of the pMG plasmid family enables an easy exchange of all features like antibiotic resistance genes (EcoRI/SalI), promoters (EcoRI/PstI), 5′ UTR (PstI/HindIII) and tags (HindIII/Acc65I) leading to a significant increase of applicabilities.
The nucleotide sequence of the B. subtilis fragments correspond with the EMBL Nucleotide Sequence Database accession number AP012496.1.
|EMBL Accession number:||AP012496.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||06/06/2013|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: EcoRI/SalI, HaeII, HincII and NotI.|
|History of deposit:||This plasmid was deposited by Dr. M. Gimpel(1) and Prof. Dr S. Brantl(1).
(1) AG Bakteriengenetik, FSU Jena, Jena, Germany
|Plasmid reference:||Gimpel et al., J. Microbiol. Methods 91 (2012), 312-317 [PMID: 22982324] [DOI: 10.1016/j.mimet.2012.09.003]
|Restricted distribution:||- BCCM MTA
- The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC.
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 DH5α|
|Host reference:||Focus 8 (1986), 9
|Related host reference:||Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml) + chloramphenicol (34 μg/ml)*|
|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.