Last data update: 24 November 2020 04:18 CET
Plasmid name: pPLc321HIFNGHS (LMBP 1444)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Human interferon γ cDNA (IFNG); mutated mature sequence|
|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|
|Parental clone:||pPLc321; pTrphIFNGHS1|
|Further information:||The way this plasmid was constructed is not exactly known; so, the nucleotide sequence between the start codon of AMP (nucleotide position 4850) and the start of the PL promoter (nucleotide position 5216) may not be correct.
pPLc321HIFNGHS contains mutated mature human interferon γ cDNA (IFNG); a unique SalI site and an extra HhaI site are created by silent mutations of amino acid 140 and 130-131 resp., the 3' UTR fragment is appox. 600 bp long.
Restriction analysis suggests the presence of a XbaI site very close to the BamHI site, that is not retrieved in the sequence file.
The nucleotide sequence of the wt human IFNG DNA corresponds with the EMBL Sequence Database accession number AM903379.1.
Other names of the plasmid are pPLΔcys and pPLc236IFγΔcys+Sal.
|EMBL Accession number:||AM903379.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||15/09/2003|
Start of the mature human IFN gamma cDNA: |-> mature hIFNG | 1 5 5' AAAAAGGGTATCGATTCC.ATG.CAG.GAC.CCA.TAT.GTA.AAA 3' Met Gln Asp Pro Tyr Val Lys Nucleotide sequence at the end of the hIFNG wt DNA: 130 140 143 5' CGA.AAA.AGG.AGT.CAG.ATG.CTG.TTT.CGA.GGT.CGA.AGA.GCA.TCC.CAG.TAA.TGGTTGTC 3' Arg Lys Arg Ser Gln Met Leu Phe Arg Gly Arg arg Ala Ser Gln Nucleotide sequence at the end of the hIFNG HS mutant: 130 140 143 5' CGA.AAG.CGC.AGT.CAG.ATG.CTG.TTT.CGA.GGT.CGA.CGA.GCA.TCC.CAG.TAA.TGGTTGTC 3' Arg Lys Arg Ser Gln Met Leu Phe Arg Gly Arg Arg Ala Ser Gln * * * * HhaI SalI *: mutated nucleotide Punctuation indicated reading frame
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Prof. Dr E. Remaut(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) 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 ΔH1Δtrp|
|Host reference:||Bernard et al., Gene 5 (1979), 59-76 [PMID: 372049]
|Related host reference:||Remaut et al., Gene 15 (1981), 81-93 [PMID: 6271633]
|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.