Last data update: 24 January 2024 16:39 CET
Plasmid name: pMG14 (LMBP 8465)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p8465.gb
(View with Genome Compiler) p8465.txt p8465.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Bacillus subtilis amylase gene (amyE); fragments Strep-tag II; N-terminal |
| Promoter: | Bacillus subtilis central glycolytic genes regulator promoter (cggR) Escherichia coli lac operon promoter |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) Kanamycin (kan; tobramycin (tob)) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Bacillus sp.; integrative |
| Parental clone: | pMG9 |
| 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 pMG9 vector. This plasmid is a modular E. coli–Bacillus shuttle vector, applicable for gene overexpression in B. subtilis via chromosomal integration. The 5' UTR of B. subtilis gapA contains a transcriptional regulator. Genes can be integrated in another site by replacing the two amyE inserts 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 EcoRI/SalI fragment of MG14 contains the kanamycin/tobramycin resistance gene aadD. When cloning a fragment downstream from the lac promoter it may be advisable to use lacIq strains in order to prevent fortuitous expression of a possibly noxious polypeptide. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726805.1. 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 LT726805.1, 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. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| 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: | Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660] Rodriguez-Quinones et al., Focus 15 (1993), 110-112 Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8] Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187] Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pMG14 (LMBP 8465) is available at BCCM/GeneCorner. This plasmid was deposited by Dr. M. Gimpel and Prof. Dr S. Brantl and was published in Gimpel et al., 2012. |
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.