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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPLc321HIFNGHS (LMBP 1444)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p1444.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human interferon γ cDNA (IFNG); mutated mature sequence
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: 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
Sequence detail:
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.
Class: Recombinant plasmid
Type: Plasmid
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
Plasmid reference: -
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)
Cultivation temperature: 28°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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