Last data update: 15 January 2021 04:14 CET
Plasmid name: pLacDN4HIFNG (LMBP 1578)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|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 (RBS) of the Escherichia coli tryptophan operon attenuator peptide (trp)|
|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|
|Further information:||pLacDN4HIFNG is derived from pLacDN1HIFNG by removing the SalI site at nucleotide position 6176 (filled in with T4 DNA polymerase), resulting in a unique SalI site between human interferon γ (hIFNG) and lacZ gene. 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.
hIFNγ is a mutated variant, containing a unique SalI site and an extra HhaI site at position 390 of the coding region.
Other name of the plasmid is pDN4G.
|EMBL Accession number:||-|
|Latest sequence update:||31/01/1989|
Nucleotide sequence at the end of the hIFNG wt gene: 131 139 140 Arg Lys Arg Ser Gln Met Leu Phe Arg Gly Arg Arg Ala Ser Gln * 5' CGA.AAA.AGG.AGT.CAG.ATG.CTG.TTT.CGA.GGT.CGA.AGA.GCA.TCC.CAG.TAA TGGTTG 3' Nucleotide sequence at the end of the hIFNG variant: 131 139 140 Arg Lys Arg Ser Gln Met Leu Phe Arg Gly Arg Pro Ser Arg Gly Cys Ser 5' CGA.AAG.CGC.AGT.CAG.ATG.CTG.TTT.CGA.GGT.CGA.CCC.TCT.AGA.GGC.TGC.AGC. HhaI SalI XbaI PstI Leu * CTC.TAG.A 3' XbaI *: Termination codon of hIFNG gene. Nucleotide sequence around the SalI site of hIFNG at the junction with lacZ: 139 AA9 of lacZ Arg Gly Arg Pro Ser Arg Ile Pro Gly Asp Pro Val Val 5' CGA.GGT.CGA.CCC.TCT.AGA.ATT.CCC.GGG.GAT.CCC.GTC.GTT 3' SalI XbaI EcoRI SmaI BamHI XmaI Punctuation indicates reading frame.
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|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
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 MC1061(λ)|
|Host reference:||Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329]
|Related host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|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.