Last data update: 24 January 2024 16:39 CET
Plasmid name: pC2HP (LMBP 9243)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p9243.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Candida albicans gDNA SV40 large T-antigen nuclear localization signal (NLS); N-terminal Viral protein 16 activation domain (VP16ad, AD VP16); N-terminal FLAG epitope tag; N-terminal |
| Promoter: | Candida albicans sulfate adenylyltransferase promoter (MET3) Phage SP6 promoter Escherichia coli lac operon promoter Escherichia coli ß-lactamase promoter (amp) |
| Ribosome binding site: |
- |
| Terminator: | Candida albicans actin terminator (ACT1) |
| Selection marker: | Neomycin (neo; kanamycin (kan)) Candida dubliniensis ARG4 (CdARG4); auxotrophic |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Candida albicans; arg(-), integrative |
| Parental clone: | pSN69 |
| Further information: | The plasmid was constructed as follows: 1) a synthetic oligonucleotide sequence, containing a SV40 NLS sequence, a 3x FLAG epitope tag sequence flanked by two glycine linkers and a multiple cloning site were cloned into the AatII/SgrAI opened pSN69 vector; 2) the C. albicans MET3 promoter and ACT1 terminator sequences were PCR amplified and cloned as AatII/BspEI and SgrAI/BstBI fragments respectively into the intermediate vector; 3) a 850 bp fragment of C. albicans genomic DNA on chromosome 2, located between genes RXT3 and ORF19.3569, was PCR amplified and cloned as a PflMI/ApaI fragment into the intermediate vector; 4) a NotI restriction site was introduced in the unique SwaI site of the intermediate vector; 5) VP16ad was PCR amplified from pMET3-VP16 and cloned into the SacII/NheI opened intermediate vector; 6) a new multiple cloning site was introduced between the StuI and EcoRI sites of the intermediate vector, introducing XmaI-AscI-MluI-ScaI sites. The plasmid can be linearised by NotI restriction to allow genomic integration. The plasmid is meant as a prey plasmid for yeast two-hybrid analysis. |
| EMBL Accession number: | - |
| Latest sequence update: | 30/04/2015 |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr P. Van Dijck(1) (2). It was constructed by Dr B. Stynen(1) (2). (1) Department of Molecular Biology, VIB, Leuven, Belgium (2) Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, Leuven, Belgium |
| Plasmid reference: | Stynen et al., Nucleic Acids Res. 38 (2010), e184 [PMID: 20719741] [DOI: 10.1093/nar/gkq725] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12xB DB3.1 |
| Host reference: | - |
| Related host reference: | Bernard et al., J. Mol. Biol. 226 (1992), 735-745 [PMID: 1324324] |
| Cultivation medium: | LB-Lennox + kanamycin (50 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | While the plasmid is provided in Escherichia coli K12xB DB3.1, use of a ccdB-resistant strain is not necessary for propagation. |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pC2HP (LMBP 9243) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Van Dijck and was published in Stynen et al., 2010. |
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.