Last data update: 24 January 2024 16:39 CET
Plasmid name: pVIK109 (LMBP 4115)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4115.gb
(View with Genome Compiler) p4115.txt p4115.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Escherichia coli alkaline phosphatase A gene (phoA); without translational start site and signal sequence |
| Promoter: | - |
| Ribosome binding site: |
- |
| Terminator: | - |
| Selection marker: | Neomycin (neo; kanamycin (kan)) |
| Replicon: | Escherichia coli plasmid R6K origin; defective pir gene Broad-host-range plasmid RK2/RP4 origin of transfer (oriT) |
| Host range: | Escherichia coli; use strains supplying the R6K pir function Gram-negative bacterial strains |
| Parental clone: | pVIK108 |
| Further information: | This plasmid was constructed as follows: 1) pVIK107 was digested with SalI and Psp1406I (AclI); 2) the resulting fragments were filled in with T4 DNA polymerase and ligated again, screening for a plasmid, pVIK108, that lacks the lacZ gene and contains the neomycin resistance gene and the R6K and RP4 derived segments as present in pVIK107. Because of the ligation, the original SalI site was restored in pVIK108; 3) the 1.5 kb XbaI-XhoI fragment from the mini-Tn5 phoA element (de Lorenzo et al. (1990)), containing the phoA gene, was inserted into the XbaI and SalI sites of pVIK108, resulting in pVIK109. The phoA gene lacks its promoter, translational start site and signal sequence. pVIK109 is a suicide vector designed to create translational fusions between chromosal or plasmid-encoded genes and the phoA reporter gene. This plasmid can be used to measure transcriptional regulation of particular promoters and to study gene regulation at the translational level. To this end, the plasmid requires the introduction of a promoter and a translation initiation region. Introduce the plasmid by electroporation, or through conjugation, using plasmid RK2/RP4 transfer functions (provided in trans by RK2/RP4, RK2/RP4-derived vectors, other replicons carrying RK2/RP4 transfer functions (e.g. pRK2073) or E. coli strains expressing the tra genes of RK2/RP4 (e.g. S17-1(λpir)). The nucleotide sequence of the mature phoA gene corresponds with the EMBL Nucleotide Sequence Database accession number M13345.1. |
| EMBL Accession number: | M13345.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 24/09/2008 |
| Sequence detail: | Nucleotide sequence of the multicloning site:
5' ... G.AAT.TCC.CGG.GAG.AGC.TCG.ATA.TCG.CAT.GCG.GTA.CCT.CTA.GAG.GAT
EcoRI SmaI SacI EcoRV SphI KpnI XbaI BamHI
CCC.CGG.GTA.CCT.GAC ... 3'
SmaI KpnI
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: PstI, XbaI and XbaI/XcmI. The restriction site analysis pattern does suppose that the plasmid is about 500 bp longer than the compiled sequence. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr S.C. Winans(1). It was constructed by Dr V.S. Kalogeraki(1). (1) Section of Microbiology, Cornell University, Ithaca, USA |
| Plasmid reference: | Kalogeraki et al., Gene 188 (1997), 69-75 [PMID: 9099861] |
| Related plasmid reference: | De Lorenzo et al., J. Bacteriol. 172 (1990), 6568-6572 [PMID: 2172217] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 SY327(λpir) |
| Host reference: | Miller et al., J. Bacteriol. 170 (1988), 2575-2583 [PMID: 2836362] |
| Related host reference: | Kalogeraki et al., Gene 188 (1997), 69-75 [PMID: 9099861] |
| Cultivation medium: | LB-Lennox + kanamycin (50 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pVIK109 (LMBP 4115) is available at BCCM/GeneCorner. This plasmid was deposited by Dr S.C. Winans and was published in Kalogeraki et al., 1997. |
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.