Last data update: 24 January 2024 16:39 CET
Plasmid name: pVIK112 (LMBP 4118)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4118.gb
(View with Genome Compiler) p4118.txt p4118.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Escherichia coli lac Z gene (lacZ); with altered 5' end Escherichia coli lac Y gene (lacY) Escherichia coli lac A gene (lacA); fragment |
| Promoter: | - |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| 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: | pVIK107 |
| Further information: | This plasmid was constructed by digesting pVIK107 with XbaI and SalI, and introducing a chemically synthesized DNA fragment containing the complementary sequences 5'-CTAGATGACTGACAGGAAACAGCTATGACCATGG-3' and 5'-TCGACCATGGTCATAGCTGTTTCCTGTCAGTCAT-3'. As a result, the original XbaI and SalI sites were reconstituted. Furthermore, translation termination codons were introduced in all three reading frames downstream from the XbaI site. The ribosome binding site encompassing the first three codons of lacZ were introduced directly upstream of the SalI site thus reconstituting a functional β-galactosidase. pVIK112 is a suicide vector designed to create transcriptional fusions between chromosal or plasmid-encoded genes and the lacZ reporter gene. This plasmid can be used to measure transcriptional regulation of particular promoters. 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 this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727310.1. |
| EMBL Accession number: | LT727310.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 22/04/2008 |
| Sequence detail: | Nucleotide sequence of the multicloning site:
* * *
5' ... GA.ATT.CCC.GGG.AGA.GCT.CGA.TAT.CGC.ATG.CGG.TAC.CTC.TAG A TGA C TGA
EcoRI SmaI SacI EcoRV SphI KpnI XbaI
CAGGAAACAGCT ATG.ACC.ATG.GTC.GAC.CTG.CAG.CCA.AGC.TTC ... 3'
----- ^ NcoI SalI PstI HindIII
^: start codon of the lacZ coding sequence.
-: ribosome binding site of the lacZ gene.
*: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoRV, PstI, SacI, SalI and XbaI. |
| 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: | Tiedeman et al., Nucleic Acids Res. 16 (1988), 3587 [PMID: 3131742] Minton, Gene 31 (1984), 269-273 [PMID: 6098531] Miller et al., J. Bacteriol. 170 (1988), 2575-2583 [PMID: 2836362] |
| 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: |
pVIK112 (LMBP 4118) 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.