Last data update: 24 January 2024 16:39 CET
Plasmid name: pSP64GalERVP (LMBP 3298)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3298.gb
(View with Genome Compiler) p3298.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Saccharomyces cerevisiae GAL4 DNA-binding domain (GALbd) Human estrogen receptor (ER) hormone binding domain Herpes simplex viral protein VP16 transcriptional activation domain (VPad, VP16ad) |
| Promoter: | Phage SP6 promoter Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pSP64 |
| Further information: | The plasmid was constructed by ligating the GAL-ER-VP fusion gene to the SacI and HindIII sites of the polylinker of pSP64, preserving the polylinker. The GAL-ER-VP gene encodes the DNA-binding domain of the Saccharomyces cerevisiae GAL4 protein fused to the hormone binding domain of the human estrogen receptor (ER). This GAL-ER fusion is converted to a transcription factor by fusion to the strong transcriptional activation domain of the Herpes simplex protein VP16 (VPad). The fusion of VP16 at the C-terminus of GAL-ER (GAL-ER-VP) seems to give better promoter induction as compared to the fusion of VP16 at the N-terminus (VP-GAL-ER). The GAL-ER-VP fusion gene can be excised as a BamHI fragment for insertion into an expression vector. When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) 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 LT727459.1. |
| EMBL Accession number: | LT727459.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 19/09/1995 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglII, HindIII, NcoI and XbaI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr S. Braselmann(1). (1) Onyx Pharma., Richmond, USA |
| Plasmid reference: | - |
| Related plasmid reference: | Braselmann et al., Proc. Natl. Acad. Sci. U.S.A. 90 (1993), 1657-1661 [PMID: 8446579] |
| Restricted distribution: | - BCCM MTA |
| 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) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSP64GalERVP (LMBP 3298) is available at BCCM/GeneCorner. This plasmid was deposited by Dr S. Braselmann. |
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.