Last data update: 24 January 2024 16:39 CET
Plasmid name: pAmp_Tn[IR-I-SceI-FRT(KanR-OriE-SacB)]_PgroEL-Transposase (LMBP 11289)
| New search | Print data sheet |
| Price category: | Cat. 2 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p11289.gb
(View with Genome Compiler) p11289.txt p11289.pdf |
| Sequence analysis results Genecorner: |
|
| Cloned DNA: |
Hyperactive Himar1 transposase cDNA (Himar1) Bacillus subtilis levansucrase cDNA (sacB, GeneID 936413) |
| Promoter: | Mycobacterium bovis molecular chaperone GroEL promoter (groEL2) |
| Ribosome binding site: |
- |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) Neomycin (neo; kanamycin (kan)) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pAmp_Tn[IR-I-SceI-FRT(KanR-OriE)]_PgroEL |
| Further information: | The plasmid was constructed by cloning the PCR amplified B. subtilis SacB coding sequence into the PmeI opened pAmp_Tn[IR-I-SceI-FRT(KanR-OriE)]_PgroEL-Transposase vector. The plasmid contains an optimized Himar1 transposon, which consists of a kanamycin resistance marker, a pMB1 origin of replication and a SacB negative selection marker, flanked by two FRT sites, two I-SceI sites and two inverted repeats (IR). FlpE mediated removal of the transposon cassette followed by sucrose selection leaves an IR-I-SceI-FRT-I-SceI-IR scar. ISceI meganuclease mediated removal of the transposon cassette followed by sucrose selection has the potential of creating KO mutants free of heterologous DNA. |
| EMBL Accession number: | - |
| Latest sequence update: | 18/08/2020 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglII/XhoI, EcoRI, HindIII, PvuII and SacII. This plasmid has also been fully sequenced but the NGS sequence data were unfortunately incomplete. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr N. Callewaert(1)(2). It was constructed by Dr K. Borgers(1)(2). (1) VIB-UGent Center for Medical Biotechnology, VIB, Ghent, Belgium (2) Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Borgers et al., mSystems 5 (2020), e00180-20 [PMID: 32788404] [DOI: 10.1128/mSystems.00180-20] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 DH5αT1R |
| Host reference: | - |
| Related host reference: | Killmann et al., J. Bacteriol. 178 (1996), 6913-6920 [PMID: 8955314] [DOI: 10.1128/jb.178.23.6913-6920.1996] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pAmp_Tn[IR-I-SceI-FRT(KanR-OriE-SacB)]_PgroEL-Transposase (LMBP 11289) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr N. Callewaert and was published in Borgers et al., 2020. |
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.