Last data update: 24 January 2024 16:39 CET
Plasmid name: pNIM1A-tTA (LMBP 10509)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p10509.gb
(View with Genome Compiler) p10509.txt p10509.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Tetracycline-responsive transcriptional activator (tTA); codon optimised for C. albicans (CatTA) Aequorea victoria green fluorescent protein DNA (GFP); codon optimised for C. albicans (CaGFP) |
| Promoter: | Candida albicans actin promoter (ACT1) Candida albicans alcohol dehydrogenase 1 promoter (ADH1) Candida albicans opaque-phase-specific protein 4 promoter (OPS4, OP4); minimal promoter with tet operator (OPS4T, OP4T) Phage T3 promoter Phage T7 gene 10 promoter (T7g10) Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | Candida albicans actin terminator (ACT1) Candida albicans ubiquinol cytochrome-c reductase subunit 7 terminator (QCR7) |
| Selection marker: | Ampicillin (amp) Nourseothricin (ntc) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin |
| Host range: | Escherichia coli Candida albicans; integrative |
| Parental clone: | pNIM1 |
| Further information: | The plasmid was constructed by replacing the CartTA coding sequence in pNIM1 with the CatTA coding sequence. The plasmid is intended for chromosomal integration of the GFP-nourseothricin resistance cassette into the C. albicans genome via recombination with the ADH1 promoter and 3' ADH1 sequence. Integration allows for tetracycline/doxycycline regulated expression of GFP via a Tet-Off transactivator, under the control of the C. albicans ADH1 promoter. The coding sequence of GFP and tTA, a fusion of the E. coli derived Tet repressor and the S. cerevisiae GAL4 activation domain, were codon optimised for C. albicans, as well as CaSAT1 which is a fusion of C. albicans ACT1 including the intron and E. coli SAT1 and confers resistance to nourseothricin. The nucleotide sequence of the plasmid corresponds with the EMBL Nucleotide Sequence Database accession number KX766187.1. |
| EMBL Accession number: | KX766187.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 05/12/2017 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, DpnI, HindIII/PvuI and NdeI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | The plasmid was deposited by Dr K. Ganesan(1). It was constructed by Dr S. Bijlani(1). (1) CSIR Institute of Microbial Technology, Chandigarh, India |
| Plasmid reference: | Bijlani et al., Curr. Genet. 64 (2018), 303-316 [PMID: 28597304] [DOI: 10.1007/s00294-017-0720-9] |
| Restricted distribution: | - BCCM MTA - The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC. |
| Distributed as: | 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 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pNIM1A-tTA (LMBP 10509) is available at BCCM/GeneCorner. The plasmid was deposited by Dr K. Ganesan and was published in Bijlani et al., 2018. |
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.