Last data update: 24 January 2024 16:39 CET
Plasmid name: pJB3kan1 (LMBP 3061)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3061.gb
(View with Genome Compiler) p3061.txt p3061.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Broad-host-range plasmid RK2 trans-acting replication initiator protein DNA (trfA) |
| Promoter: | Escherichia coli lac operon promoter Transposon Tn5 neomycin resistance gene promoter (neo) |
| Ribosome binding site: |
- |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) Neomycin (neo; kanamycin (kan)) |
| Replicon: | Broad-host-range plasmid RK2 vegetative replication origin Broad-host-range plasmid RK2/RP4 origin of transfer (oriT) |
| Host range: | Escherichia coli Gram-negative bacterial strains |
| Parental clone: | pJB3 |
| Further information: | The plasmid was constructed by inserting the kanamycin resistance Genblock… (Pharmacia) as an EcoRI fragment into the unique EcoRI site of pJB3. The orientation of the inserted fragment is anti-clockwise. This IncP plasmid is used as a broad-host-range vector. The plasmid RK2 trfA gene encodes two polypeptide products P382 and P285 of 382 and 285 amino acids respectively, using a different initiation codon within the same reading frame. The resulting protein is essential for vegetative replication. When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide. 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 strains expressing the tra genes of RK2/RP4). The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727430.1. The nucleotide sequence of the pJB3 vector corresponds with the EMBL Nucleotide Sequence Database accession number U73899.1. Other name of the plasmid is pJB3Kan1. |
| EMBL Accession number: | U73899.1, view at EMBL, GenBank, DDBJ LT727430.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 21/03/1995 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: HindIII, XbaI and XmnI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr E. Remaut(1) (2). It was constructed by K. Keymeulen(2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Blatny et al., Appl. Environ. Microbiol. 63 (1997), 370-379 [PMID: 9023917] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pJB3kan1 (LMBP 3061) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr E. Remaut . |
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.