Last data update: 24 January 2024 16:39 CET
Plasmid name: pPP1IVHAsi (LMBP 3924)
| New search | Print data sheet |
| 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] |
| Related plasmid reference: | Manual of Pichia Expression (Invitrogen) |
| 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: | Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660] Rodriguez-Quinones et al., Focus 15 (1993), 110-112 Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8] Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187] Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pPP1IVHAsi (LMBP 3924) is available at BCCM/GeneCorner. This plasmid was deposited by Dr X. Saelens and Prof. Dr W. Min Jou and was published in Saelens et al., 1999. |
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.