Last data update: 24 January 2024 16:39 CET
Plasmid name: pJBtrc10 (LMBP 3523)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3523.gb
(View with Genome Compiler) p3523.txt p3523.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Escherichia coli lac repressor gene (lacI) Broad-host-range plasmid RK2 trans-acting replication initiator protein DNA (trfA) |
| Promoter: | Escherichia coli hybrid tryptophan/lacUV5 promoter (trc) Escherichia coli lac repressor promoter; mutant (lacI(q)) Phage T3 promoter Transposon Tn5 neomycin resistance gene promoter (neo) |
| Ribosome binding site: |
Ribosome binding site (RBS) of the phage T7 gene 10 (T7g10) |
| Terminator: | Phage fd terminator Phage T7 gene 10 terminator (T7g10) |
| Selection marker: | Tetracycline (tet) |
| 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: | pJBtrc2; pLT10T3 |
| Further information: | The plasmid was constructed as follows: first, pLT10T3 was digested with ApaI and ligated to eliminate a 89 bp fragment that contains a XbaI site; then, the XbaI/BsaAI fragment of this intermediate construct, containing the T7 gene 10 RBS, the T3 promoter, two T7 gene 10 terminators and two fd terminators, was inserted between the XbaI and ScaI sites of pJBtrc2. 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. This IncP plasmid is used as a broad-host-range expression vector. 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 LT727349.1. |
| EMBL Accession number: | LT727349.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 22/01/2001 |
| Sequence detail: | Nucleotide sequence following the Shine-Dalgarno (SD) position of the T7 gene 10 ribosome binding site:
...TATAATGTGTGG| AATTGTGAGCGGATAACAATTTCACTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATA
trc promoter->| XbaI ------
SD
TACAT.ATG.GGC.CCG.GAT.CCC.TCG.AGG.TCG.ACG.AAT.TCG.AGC.TCG...
***
NdeI ApaI XhoI SacI
***: start codon. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: ApaI, BglI, SspI and XbaI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr E. Remaut(1) (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: | Brosius et al., J. Biol. Chem. 260 (1985), 3539-3541 [PMID: 2579077] |
| 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 + tetracycline (10 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pJBtrc10 (LMBP 3523) 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.