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GeneCorner plasmid details

Last data update: 22 October 2020 04:25 CEST

Plasmid name: pAJ401mds1 (LMBP 4485)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p4485.gb (View with Genome Compiler)
p4485.txt
p4485.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Trichoderma reesei 1,2-α-D-mannosidase cDNA (mds1)
Promoter: Saccharomyces cerevisiae phosphoglycerate kinase promoter (PGK)
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Saccharomyces cerevisiae phosphoglycerate kinase terminator (PGK)
Selection marker: Ampicillin (amp)
Saccharomyces cerevisiae URA3; auxotrophic
Replicon: Escherichia coli plasmid pMB1 origin
Saccharomyces cerevisiae 2 micron plasmid origin (2μ)
Host range: Escherichia coli
Saccharomyces cerevisiae; ura3(-)
Parental clone: pAJ401
Further information: This plasmid was isolated from a T. reesei QM9414 cDNA library obtained from A. Saloheimo and Dr M. Penttil“ (VTT Biotechnology and Food Research, Espoo, Finland). The library was made by inserting the T. reesei cDNA into the EcoRI-XhoI opened pAJ401 vector. The pAJ401 vector was derived from pFL60 (Minet et al., 1990) by interchanging EcoRI and XhoI cloning sites.
pAJ401mds1 is a yeast multicopy plasmid that contains the T. reesei mds1 cDNA.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727223.1.The nucleotide sequence of pFL60 was obtained from A. Saloheimo.
The nucleotide sequence of the PGK promoter corresponds with the PGK1 upstream sequence in the Saccharomyces Genome Database (SGD), except for the 'G' nucleotide at promoter position 707 that is a 'C' in the SGD and the 'A' nucleotide at promoter position 819 that is absent in the SGD.
The nucleotide sequence of the PGK terminator corresponds with the EMBL Nucleotide Sequence Database accession number J01342.1.
The nucleotide sequence of the T. reesei mds1 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF212153.1. Be aware that the resulting amino acid sequence differs from the one described by Maras et al. (2000). Furthermore, additional amino acid changes have been noticed by a BCCM/GeneCorner customer (02/09/02). Some of the latter changes could be confirmed by the latest sequence analysis results obtained with this GeneCorner plasmid by Dr Maras (11/03/03), others couldn't. Referring to the customer's sequence analysis, mds1 codon 116 encodes for isoleucine (I) instead of serine (S); consequently, the BamHI site at this position is absent. The absence of the BamHI site was confirmed by restriction site analysis at BCCM/GeneCorner, however the nucleotide sequence was not adapted.
Other name of the plasmid is pAJ401mannosidase.
EMBL Accession number: AF212153.1, view at EMBL, GenBank, DDBJ
J01342.1, view at EMBL, GenBank, DDBJ
LT727223.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 09/10/2003
Sequence detail:
Overview of the amino acid differences of the mds1 coding sequence:

AA position     EMBL     Maras2000     Customer     Maras2003

116             S        S             I            S
292             G        G             A            A
298             G        G             A            A
301             G        G             A            A
332             G        G             A            G
361             G        G             A            G
364             T        N             N            N
438             L        F             F            F
496             T        N             N            N
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: AflIII, BglII, ClaI, NcoI, SphI, XbaI and XhoI.

The 512 bp BglII fragment is slightly larger as compared to the 516 bp ClaI fragment, suggesting that one of the BglII sites at the 3' end of the mds1 cDNA insert (positions 3400 and 3444) is not present.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr M. Maras(1) (2) and Prof. Dr R. Contreras(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Maras et al., J. Biotechnol. 77 (2000), 255-263 [PMID: 10682284]
Related plasmid reference: Minet et al., Curr. Genet. 18 (1990), 287-291 [PMID: 2253271]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061
Host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Related host reference: Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
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:

pAJ401mds1 (LMBP 4485) is available at BCCM/GeneCorner. This plasmid was deposited by Dr M. Maras and Prof. Dr R. Contreras and was published in Maras et al., 2000.

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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