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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMa254 (LMBP 2099)

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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: -

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.

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