Last data update: 24 January 2024 16:39 CET
Plasmid name: pIMAY-Z (LMBP 10259)
| New search | Print data sheet |
| Price category: | Cat. 3 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p10259.gb
(View with Genome Compiler) p10259.txt p10259.pdf |
| Sequence analysis results Genecorner: |
NGS: d12-gc-may2021.fasta |
| Cloned DNA: |
Escherichia coli lac Z gene (lacZ) Tetracycline repressor (TetR) Secretion regulation gene Y antisense RNA (secY) |
| Promoter: | Phage T3 promoter Phage T7 gene 10 promoter (T7g10) Staphylococcus tetracycline-regulated hybrid promoter (xyl/tetO) |
| Ribosome binding site: |
- |
| Terminator: | - |
| Selection marker: | Chloramphenicol (cam) |
| Replicon: | Escherichia coli plasmid p15A origin Broad-host-range plasmid RK2/RP4 origin of transfer (oriT) Broad-host-range plasmid pWV01 origin; temperature sensitive variant (pVE6007) |
| Copy number: | Low copy number |
| Host range: | Escherichia coli Staphylococcus aureus |
| Parental clone: | pIMAY |
| Further information: | The plasmid was constructed by cloning the lacZ coding sequence into the pIMAY plasmid via sequence- and ligase-independent cloning (SLICE). pIMAY-Z is meant for allelic exchange in Staphylococcus aureus. The plasmid contains the secY antisense sequence. Anhydrotetracycline-mediated induction of secY antisense transcripts via the xyl/tetO promoter, a condition that is not compatible with staphylococcal growth, selects for chromosomal excision and loss of plasmid. Spontaneous mutations may arise in the pWV01 origin of replication when maintained as a plasmid-carrying E. coli culture at room temperature. The nucleotide sequence of the parental pIMAY vector corresponds with the EMBL Nucleotide Sequence Database accession number JQ621981.1. |
| EMBL Accession number: | JQ621981.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 27/01/2017 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglII, EcoRV, NcoI/NdeI and SpeI. This plasmid has also been fully sequenced. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr I. Monk(1). (1) Department of Microbiology and Immunology, Doherty Institute, University of Melbourne, Melbourne, Australia |
| Plasmid reference: | Monk et al., MBio 6 (2015), e00308-15 [PMID: 26015493] [DOI: 10.1128/mBio.00308-15] |
| Related plasmid reference: | Maguin et al., J. Bacteriol. 174 (1992), 5633-5638 [PMID: 1324906] Bae and Schneewind, Plasmid 55 (2006), 58-63 [PMID: 16051359] [DOI: 10.1016/j.plasmid.2005.05.005] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | 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 + chloramphenicol (34 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pIMAY-Z (LMBP 10259) is available at BCCM/GeneCorner. This plasmid was deposited by Dr I. Monk and was published in Monk et al., 2015. |
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.