LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMcT4HTNFd31-35iGSG (LMBP 2621)

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


Cloned DNA: Human tumor necrosis factor cDNA (TNF); mutated sequence
Promoter: Phage T4 gene 32 promoter (T4g32)
binding site:
Ribosome binding site (RBS) of the phage T4 gene 32 (T4g32)
Terminator: Phage fd terminator
Escherichia coli tryptophan operon terminator (trp); synthetic sequence
Selection marker: Chloramphenicol (cam)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Parental clone: pMa-T4-hTNF-TA
Further information: This plasmid was constructed by ss mutagenesis, where amino acids 31 to 35 were replaced by GGT.TCA.GGC
This mutagenesis created a unique StuI restriction site, eliminated the unique XcmI site of pMa-T4-hTNF-TA, and also
(coding for Gly-Ser-Gly).
eliminated a BglI site, which now becomes unique.
After mutagenesis, use mutS strains for primary transformation (e.g. sup(-) strain WK6mutS, sup(+) strain BMH71-18mutS); for segregation of possible mutants: sup(-) strains (e.g. WK6).
The nucleotide sequence of the hTNF cDNA corresponds with the EMBL Nucleotide Sequence Database accession number X01394.1.
Other name of the plasmid is pMcT4HTNFTmdel31-35+GSG.
EMBL Accession number: X01394.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 11/01/1993
Sequence detail:
Overview of he deletion and insert of hTNF in pMcT4HTNFT

SN : 2346       pMcT4TNFT (wildtype human TNF)

     26  27  28  29  30  31  32  33  34  35  36  37  38   
     Leu Gln Trp Leu Asn Arg Arg Ala Asn Ala Leu Leu Ala
         AlwNI             NciI   XcmI   BglI     BalI
                                                 EaeI BstXi

SN: 2621 pMcT4TNFd31-35iGSG

     26  27  28  29  30              36  37  38   
     Leu Gln Trp Leu Asn Gly Ser Gly Asn Ala Leu Leu Ala
         AlwNI                 StuI               BalI
                                                 EaeI BstXi
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 R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Van Ostade et al., Eur. J. Biochem. 220 (1994), 771-779 [PMID: 8143731]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 SURE
Host reference: Greener, Strategies 3 (1990), 5-6
Cultivation medium: LB-Lennox + chloramphenicol (25 μg/ml)
Cultivation temperature: 37°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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