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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: ppdi100 (LMBP 3050)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: ppdi100.gb (View with Genome Compiler)
ppdi100.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Rat protein disulfide isomerase cDNA (rPDI)
Promoter: -
Ribosome
binding site:
-
Terminator: -
Selection marker: Tetracycline (tet)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Parental clone: pBR322; λpdi6; ppdi306
Further information: This plasmid was constructed as follows: the NcoI-HindII fragment from λpdi6 was ligated to the NcoI-ScaI vector fragment from ppdi306. λpdi6 contains a large fragment from the 3' part of the rat protein disulfide isomerase cDNA (rPDI), isolated from a rat liver cDNA library, in λgt10; ppdi306 contains a 280 bp insert with most of the 5' sequences of the rPDI cDNA, constructed by insertion of a cDNA fragment that was also isolated from a rat liver cDNA library into the PstI-digested pBR322 via dC/dG-tailing. As a result, ppdi100 contains the complete rPDI cDNA.
The entire rPDI insert is excisable using ApaI at the 5' end and SmaI at the 3' end.
The availability of the rPDI cDNA, after insertion in an expression clone, allows direct analysis of the role of PDI in the process of disulfide bond interchange.
Since the ampicillin resistance gene has been disrupted by the rPDI insert, ppdi100 only confers resistance to tetracycline.
The nucleotide sequence of the rPDI cDNA corresponds with the EMBL Nucleotide Sequence Database (Release 36; Accession number X02918).
The nucleotide sequence at the 5' end and at the 3' end of the rPDI cDNA insert is not exactly known. Neither is the length of the dC/dG-tails.
EMBL Accession number: X02918, view at EMBL, GenBank, DDBJ
Latest sequence update: 13/06/1994
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: ApaI/SmaI, EcoRI, HindII/NcoI, NheI/StyI and SpeI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr W.J. Rutter(1).
(1) University of California, San Francisco, USA
Plasmid reference: Edman et al., Nature 317 (1985), 267-270 [PMID: 3840230]
Related plasmid reference: De Sutter et al., Gene 141 (1994), 163-170 [PMID: 8163184]
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 + tetracycline (10 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
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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