Last data update: 24 January 2024 16:39 CET
Plasmid name: pCB1003 (LMBP 8050)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p8050.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
- |
| Promoter: | Aspergillus nidulans tryptophan C promoter (trpC) Escherichia coli lac operon promoter |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) Hygromycin (hyg) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Magnaporthe sp. Ascomycete fungi |
| Parental clone: | pCSN43; pUC19 |
| Further information: | The plasmid was constructed as follows: 1) the coding sequence of the hygromycin resistance gene from pCSN43 was mutated through three rounds of megaprimer mutagenesis, eliminating the EcoRI (GAgTTC), PstI (CTGgAG), SstII (CCGCGc) and BamHI (GGtTCC) restriction sites and eliminating the ClaI site near the start codon. The latter mutation changed the Kozak sequence from ATCGATATG to ATCcAaATG. 2) the mutated hygromycin cds from pCSN43 was PCR amplified, adding EcoRI, SalI and HpaI restriction sites at both ends. 3) the amplified hygromycin cds was ligated as an EcoRI fragment into the EcoRI site of pUC19. |
| EMBL Accession number: | - |
| Latest sequence update: | 14/01/2013 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, EcoRI, NdeI and PvuI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr K. McCluskey(1). (1) FGSC, University of Missouri, Kansas City, USA |
| Plasmid reference: | Caroll et al., Fungal Genet. Newsl. 41 (1994), 22 |
| Related plasmid reference: | Sweigard et al., Fungal Genet. Newsl. 44 (1995), 52-53 |
| 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: |
pCB1003 (LMBP 8050) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr K. McCluskey and was published in Caroll et al., 1994. |
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.