Last data update: 24 January 2024 16:39 CET
Plasmid name: pHSG415 (LMBP 3333)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3333.gb
(View with Genome Compiler) p3333.txt p3333.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Escherichia coli plasmid pSC101 replication protein A (repA); mutated sequence |
| Promoter: | - |
| Ribosome binding site: |
- |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) Chloramphenicol (cam) Neomycin (neo; kanamycin (kan)) |
| Replicon: | Plasmid pSC101 origin derived from R6-5; temperature sensitive |
| Copy number: | Low copy number |
| Host range: | Escherichia coli |
| Parental clone: | pHSG412; pACYC184 |
| Further information: | 8 different restriction endonucleases are useful for cloning: EcoRI, XhoI, XmaI, HindIII, BamHI, HindII, PstI and BstEII. All these sites are unique and, with exception of BamHI and BstEII, are located in an antibiotic resistance gene. The plasmid contains a temperature sensitive (ts) pSC101 origin, controlled by a mutated repA gene. As a consequence, it can be stably maintained in the host at temperatures below 30°C, but is rapidly lost at temperatures above 37°C. The nucleotide sequence of this plasmid was compiled according to Hashimoto-Gotoh et al. (1981) and to Hashimoto-Gotoh et al. (1979), and contains some uncertainties. The nucleotide sequence of the pSC101 origin and of the wt repA gene correspond with the EMBL Nucleotide Sequence Database accession number K00042.1. |
| EMBL Accession number: | K00042.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 10/08/1995 |
| Sequence detail: | Nucleotide sequence around the mutation of the repA gene:
56
wild type: 5' ... CAA TAC GCT CAG ... 3'
Ala
ts mutant: 5' ... CAA TAC GTT CAG ... 3'
^
Val
^: mutated nucleotide. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: AflIII, BglI/XhoI, EcoRI and ScaI. The restriction site analysis pattern did not completely fit the expected plasmid length and fragment sizes deduced from the compiled nucleotide sequence: the plasmid length seems to be rather 7500 bp to 8000 bp. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr K.N. Timmis(1). (1) Department of Microbiology, GBF, Braunschweig, Germany |
| Plasmid reference: | Hashimoto-Gotoh et al., Gene 16 (1981), 227-235 [PMID: 6282694] |
| Related plasmid reference: | Cohen et al., J. Bacteriol. 132 (1977), 734-737 [PMID: 334752] Hashimoto-Gotoh et al., J. Bacteriol. 139 (1979), 597-607 [PMID: 378979] |
| 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) + chloramphenicol (25 μg/ml) + kanamycin (50 μg/ml)* |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Cultivation remark: | *: selection of transformants on chloramphenicol; subsequent cultivation of a single colony in liquid medium with ampicillin and kanamycin. |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pHSG415 (LMBP 3333) is available at BCCM/GeneCorner. This plasmid was deposited by Dr K.N. Timmis and was published in Hashimoto-Gotoh et al., 1981. |
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.