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LMBP plasmid details

Last data update: 26 September 2020 04:22 CEST

Plasmid name: pLacDN5HIFNG (LMBP 1850)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p1850.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human interferon γ cDNA (IFNG)
Escherichia coli lac Z gene (lacZ); starting at the 9th codon
Escherichia coli lac Y gene (lacY); fragment
Promoter: Phage λ major leftward promoter (λ PL)
Ribosome
binding site:
Ribosome binding site (RBS) of the Escherichia coli tryptophan operon attenuator peptide (trp)
Terminator: -
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; use strains with a cI function, cIts for PL controlled expression
Parental clone: pLacDN3HIFNG; pPLcZCY1
Further information: pLacDN5HIFNG is derived from pLacDN3HIFNG by inserting the SacI-PstI fragment from pPLcZCY1 into the unique SacI and unique PstI sites. Due to this construction, the lacA gene is deleted and only part of the lacY is maintained.
pLacDN5HIFNG contains an in-phase fusion protein: human interferon γ/β-galactosidase.
hIFNγ is a mutated variant, containing a unique SalI site and an extra HhaI site at position 390 of the coding region. The chromosomal lacZ gene must be inactivated for detection of β-galactosidase activity of the hIFNG-lacZ fusion protein.
The natural EcoRI site near the end of lacZ was removed by mutation.
The incomplete lacY gene is still functional, but activity is lower than that of the wild type lacY. So, β-galactosidase activity measurement by X-Gal coloration is much slower.
Other name of the plasmid is pDN5G.
EMBL Accession number: -
Latest sequence update: 01/02/1989
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by E. Sablon(1) and Prof. Dr E. Remaut(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: PhD thesis Erwin Sablon (1990)
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061[pcI857]
Host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Helper plasmid: pcI857
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml)
Cultivation temperature: 28°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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