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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPP1IVHAsi (LMBP 3924)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p3924.gb (View with Genome Compiler)
p3924.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Saccharomyces cerevisiae α-mating factor 1 gene (MFα1); prepro secretion signal sequence (ppMF)
Influenza A virus (A/Victoria/3/1975(H3N2)) haemagglutinin cDNA (HA, IVHA); N-terminal anchor free
Promoter: Pichia pastoris alcohol oxidase 1 promoter (AOX1)
Ribosome
binding site:
-
Terminator: Pichia pastoris alcohol oxidase 1 terminator (AOX1)
Selection marker: Ampicillin (amp)
Pichia pastoris HIS4; auxotrophic
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Pichia pastoris; his4(-), integrative
Parental clone: pUCIVHAsm; pPIC92
Further information: This plasmid was constructed as follows: pUCIVHAsm was linearized with BamHI, filled in with T4 DNA polymerase and dNTP's and ligated with the previously hybridized oligonucleotides 5'-TAAGCGGCCGCG-3' and 5'-CGCGGCCGCTTA-3'. A clone with the NotI site downstream from the introduced TAA termination codon was chosen for further use. Then this plasmid was linearized with ApaI, flush-ended with T4 DNA polymerase and dNTP's, and subsequently digested with NotI. The mature A/Victoria/3/75 influenza haemagglutinin cDNA encoding a secreted protein (IVHAs) fragment was ligated in the NaeI-NotI opened pPIC92 plasmid.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727289.1.
EMBL Accession number: LT727289.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 08/03/1999
Sequence detail:
Nucleotide sequence at the start of the mature haemagglutinin cDNA (IVHA):

      ------ ppMF ------->        |       ||       ||-->   mature IVHA 
5' ... GGG.GTA.TCT.CTC.GAG.AAA.AGA|GAG.GCT||GAA.GCC||CAA.GAC.CTT.CCA.GGA.AAT.GAC ... 3'
       Gly Val Ser Leu Glu Lys Arg|Glu Ala||Glu Ala||Gln Asp Leu Pro Gly Asn Asp
                   XhoI           |       ||       ||

| : Kex2 cleavage site.
||: Dipeptidase cleavage.


Nucleotide sequence at the 3' end of IVHA:

                   -> IVHA
5' ... TAC.AAA.GAC.TGG.ATC.TAA.GCGGCCGC … 3'
       Tyr Lys Asp Trp Ile +++ NotI
                                XmaIII     
                           
+++: Termination codon.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglI, HindII, NcoI, SpeI and XhoI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr X. Saelens(1) (2) and Prof. Dr W. Min Jou(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Saelens et al., Eur. J. Biochem. 260 (1999), 166-175 [PMID: 10091596]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
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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