Last data update: 24 January 2024 16:39 CET
Plasmid name: pSALSOD1 (LMBP 5017)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p5017.gb
(View with Genome Compiler) p5017.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Geobacillus stearothermophilus manganese superoxide dismutase cDNA (MnSOD) |
| Promoter: | Saccharomyces cerevisiae hybrid UDP-glucose 4-epimerase/iso-1-cytochrome C promoter (GAL10/CYC1) |
| Ribosome binding site: |
- |
| Terminator: | Saccharomyces cerevisiae 2 micron plasmid (2μ) FLP terminator |
| Selection marker: | Ampicillin (amp) Saccharomyces cerevisiae URA3; auxotrophic LEU2 defective (leu2-d; auxotrophic) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Saccharomyces cerevisiae 2 micron plasmid origin (2μ); incl. region conferring stability (STB) |
| Host range: | Escherichia coli Saccharomyces cerevisiae; ura3(-) |
| Parental clone: | pEMBLyex4 |
| Further information: | The plasmid was constructed by inserting a 730 bp SmaI-BalI fragment, containing the Geobacillus stearothermophilus manganese superoxide dismutase (MnSOD) cDNA, into the SmaI opened pEMBLyex4 vector. The MnSOD cDNA is placed under control of the galactose-inducible Saccharomyces cerevisiae hybrid UDP-glucose 4-epimerase/cytochrome C1 (GAL10/CYC1) promoter and the S. cerevisiae 2 micron plasmid (2μ) FLP terminator. The GAL10/CYC1 hybrid promoter sequence consists of the upstream activation sequence of the GAL10 gene (UAS-G), the GAL10 promoter and the 5' untranslated leader sequence of the CYC1 gene. pSALSOD1 can be used as a high-copy number extrachromosomal vector using the S. cerevisiae 2 micron (2μ) plasmid origin, but it can also be directed to integrate into the yeast chromosome at LEU2 (by linearising with BstXI, EcoRI, AflII or BstEII) or at URA3 (by linearising with ApaI or StuI). pSALSOD1 contains the replication origin of the single-stranded DNA phage f1 (clockwise orientated), so that it can be rapidly switched between the plasmid and the phage mode (ssDNA) of replication. The latter requires infection with a helper phage, e.g. M13KO7. The defective LEU2 gene (leu2-d) is poorly expressed, because there are only 30 bp of upstream sequence left in this plasmid; the expression is probably controlled by a promoter within the 2μ sequences. |
| EMBL Accession number: | M26646, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 01/09/2005 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglI, BglI/SmaI, EcoRI/PstI, HindIII, NcoI/PvuI, SmaI, SphI/StuI and XmnI. As compared to the compiled nucleotide sequence, the restriction site analysis pattern revealed that the SmaI site at position 5546 could not be experimentally confirmed. As the compiled nucleotide sequence has not been adapted accordingly, SmaI is still indicated as a unique site. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr D. Inzé(1) (2) and constructed by C. Bowler(1) (2). (1) Department of Plant Systems Biology, VIB, Ghent, Belgium (2) Department of Plant Systems Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Bowler et al., J. Bacteriol. 172 (1990), 1539-1546 [PMID: 2407726] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSALSOD1 (LMBP 5017) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr D. Inzé and constructed by C. Bowler and was published in Bowler et al., 1990. |
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.