Last data update: 24 January 2024 16:39 CET
Plasmid name: pMa254 (LMBP 2099)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
pma254.gb
(View with Genome Compiler) pma254.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
- |
| Promoter: | - |
| Ribosome binding site: |
- |
| Terminator: | Phage fd terminator |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pMa58 |
| Further information: | The plasmid was designed for site-specific mutagenesis as the mutated sequence can be physically linked with a selectable marker (chloramphenicol). The plasmid provides resistance to ampicillin, but is sensitive to chloramphenicol. More than one mutagenesis round is possible via switching from ampicillin resistance to chloramphenicol resistance. pMa254 contains the replication origin of the single-stranded DNA phage f1 (anticlockwise orientated), so that it can be rapidly switched between the plasmid and the phage mode (ssDNA) of replication. The latter requires infection with a helper phage, e.g. M13KO7. The f1 part of the plasmid contains the f1 origin but also the sequence around the gene II promoter. The two trp terminators at the end of the chloramphenicol gene are not present. After mutagenesis, use mutS strains for primary transformation (e.g. sup(-) strain WK6mutS, sup(+) strain BMH71-18mutS); for segregation of possible mutants: sup(-) strains (e.g. WK6). The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727620.1. |
| EMBL Accession number: | LT727620.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 10/11/1989 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglI, HindII, MspA1I and ScaI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr P. Stanssens(1). (1) Corvas International N.V., Ghent, Belgium |
| Plasmid reference: | - |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 K514(λ) |
| Host reference: | Zabeau et al., EMBO J. 1 (1982), 1217-1224 [PMID: 6327257] |
| 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: |
pMa254 (LMBP 2099) is available at BCCM/GeneCorner. This plasmid was deposited by Dr P. Stanssens. |
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.