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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pMa-T4-hTNF-TA (LMBP 2345)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p2345.gb (View with Genome Compiler)
p2345.txt
p2345.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human tumor necrosis factor cDNA (TNF); mature sequence
Promoter: Phage T4 gene 32 promoter (T4g32)
Ribosome
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: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Parental clone: pMa58; pPLT4HTNFST8
Further information: This plasmid was constructed by inserting the EcoRI fragment from pPLT4HTNFST8, containing the phage T4 promoter, the hTNF mature sequence and the synthetic trpA terminator, clockwise into the unique EcoRI site of pMa58.
The unique SalI site of the polylinker of pMa58 was deleted.
Although the PstI restriction site of the ampicillin resistance gene was removed, the plasmid conserves its resistance to ampicillin, but it is sensitive to chloramphenicol (in sup(-) host strains).
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 this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727505.1.
The nucleotide sequence of the hTNF cDNA corresponds with the EMBL Nucleotide Sequence Database accession number X01394.1.
Other name of the plasmid is pMaT4hTNFT.
EMBL Accession number: X01394.1, view at EMBL, GenBank, DDBJ
LT727505.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 04/02/1992
Sequence detail:
Nucleotide sequence at the 5' end of the mature human TNF cDNA:

  T4 promoter + RBS -->|          |--> mature human TNF
5' ... AAATTAAAAAGGAAAT|CGATACTACT.ATG.GTC.AGA.TCA.TCT.TCT.CGA.ACC.CCG. ... 3'
                     ClaI          Met Val Arg ser Ser Ser Arg Thr Pro
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: AvaI, BstEII, EcoRI and PstI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr F. Van Roy(1) (2). It was constructed by X. Van Ostade(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., FEBS Lett. 238 (1988), 347-352 [PMID: 3049162]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH1(λ)
Host reference: Bachmann, in 'Escherichia coli and Salmonella: Cellular and Molecular Biology', American Society for Microbiology, Washington DC (1987), 1190-1219
Cultivation medium: LB-Lennox + ampicillin (100 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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