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LMBP plasmid details

Last data update: 25 September 2020 04:24 CEST

Plasmid name: pLacDN4HIFNGmb (LMBP 1776)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p1776.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)
Escherichia coli lac A gene (lacA)
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: pLacDN4HIFNG; pUC19
Further information: The plasmid was constructed by replacing the SalI-SmaI fragment from the linker of pLacDN4HIFNG by the SalI-BamHI (blunted with T4 DNA polymerase) fragment from the polylinker of pUC19.
This plasmid bears an in-phase nonsense (amber) codon in the hIFNG-lacZ fusion protein. This amber codon is very well suppressible. The chromosomal lacZ gene must be inactivated for detection of β-galactosidase activity of the hIFNG-lacZ fusion protein.
Other names of the plasmid are pDN4G2 and pLacDN4HIFNG2.
EMBL Accession number: -
Latest sequence update: 26/10/1989
Sequence detail:
Nucleotide sequence around the hIFNG-lacZ junction:

           139    amber                    AA9 of lacZ
5' CGA.GGT.CGA.CTC.TAG.AGG.ATC.GGG.GAT.CCC.GTC.GTT 3'
        SalI    XbaI            BamHI      


Punctuation indicates reading frame.
Nonsense (amber) codon.
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 DH5α[pcI857]
Host reference: Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
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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