Last data update: 24 January 2024 16:39 CET
Plasmid name: pC2HB (LMBP 9241)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p9241.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Candida albicans gDNA SV40 large T-antigen nuclear localization signal (NLS); N-terminal Staphylococcus aureus LexA repressor cDNA (LexA, lexA DNA-binding domain, lexA DBD, lexAbd, GeneID 3920198); N-terminal Influenza HA epitope encoding the haemagglutinin tagging peptide; N-terminal |
| Promoter: | Candida albicans sulfate adenylyltransferase promoter (MET3) Phage SP6 promoter Phage T7 gene 10 promoter (T7g10) Escherichia coli lac operon promoter Escherichia coli β-lactamase promoter (amp) |
| Ribosome binding site: |
- |
| Terminator: | Candida albicans actin terminator (ACT1) |
| Selection marker: | Neomycin (neo; kanamycin (kan)) Candida maltosa LEU2 (CmLEU2); auxotrophic |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Candida albicans; leu2(-), integrative |
| Parental clone: | pSN40 |
| Further information: | The plasmid was constructed as follows: 1) a synthetic oligonucleotide sequence containing a multiple cloning site and a HA-tag was cloned as a BspEI/SalI fragment into the pSN40 plasmid; 2) the S. aureus lexA gene was PCR amplified from plasmid Cip-LexA, adding the SV40 NLS sequence via the forward PCR primer, and cloned into the intermediate vector; 3) the C. albicans MET3 promoter and ACT1 terminator sequences were PCR amplified and cloned as AatII/BspEI and SalI/DraIII fragments respectively into the intermediate vector; 4) a 1104 bp fragment of C. albicans genomic DNA on chromosome 1, located between genes XOG1 and HOL1, was PCR amplified and cloned as a PvuI/SnaBI fragment into the intermediate vector; 5) the unique NdeI restriction site in the gDNA fragment was replaced by NotI and 6) a new multiple cloning site was introduced at the AscI site of the intermediate vector, introducing NheI-StuI-SfuI-AscI-MluI-ScaI sites. The plasmid is meant as a bait plasmid for yeast two-hybrid analysis. The plasmid can be linearised by NotI restriction to allow genomic integration. The nucleotide sequence of the S. aureus lexA cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AY033082.1. |
| EMBL Accession number: | AY033082.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 18/06/2014 |
| 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: |
pC2HB (LMBP 9241) 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.