Last data update: 24 January 2024 16:39 CET
Plasmid name: pKDsgRNA-FRT (LMBP 11104)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p11104.gb
(View with Genome Compiler) p11104.txt p11104.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Single-guide RNA targeting the flippase recognition target site (FRT) Escherichia coli plasmid pSC101 replication protein A (repA); mutated sequence Escherichia coli DNA-binding transcriptional regulator AraC (araC) Escherichia coli host-nuclease inhibitor protein Gam (gam) Escherichia coli phage recombination protein Bet (bet) Escherichia coli exonuclease (exo) Histidine tag (His-tag) Myc epitope |
| Promoter: | Escherichia coli β-lactamase promoter (amp) Escherichia coli L-arabinose operon promoter (araB, araBAD, BAD) Escherichia coli tetracycline inducible promoter (PLtetO-1) |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli ampicillin resistance gene (amp) |
| Terminator: | Escherichia coli rrnB operon T1 terminator Phage λ rho-independent major leftward terminator (λ tL3) |
| Selection marker: | Streptomycin (Sm; spectinomycin (Sp)) |
| Replicon: | Plasmid pSC101 origin derived from R6-5; temperature sensitive |
| Copy number: | Low copy number |
| Host range: | Escherichia coli |
| Parental clone: | pKDsgRNA-ack |
| Further information: | The plasmid was constructed by cloning the sgRNA for FRT targeting into the pKDsgRNA-ack vector via Gibson assembly. The plasmid contains the arabinose inducible lambda red recombineering genes (gam, bet, exo). It can be used for anhydrotetracycline inducible sgRNA-FRT expression. The plasmid carries the aadA gene which confers resistance to spectinomycin and streptomycin. |
| EMBL Accession number: | - |
| Latest sequence update: | 14/07/2022 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoRI, EcoRV/HincII, PvuI and XbaI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr T. Swings and Prof. Dr J. Michiels(1). (1) Centre of Microbial and Plant Genetics, Department of Microbial and Molecular Systems, KU Leuven, Leuven, Belgium |
| Plasmid reference: | Swings et al., Nat. Commun. 9 (2018), 2231 [PMID: 29884781] [DOI: 10.1038/s41467-018-04651-5] |
| Restricted distribution: | - BCCM MTA - The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC. |
| 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 + spectinomycin (50 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | Addgene 113398 |
Refer in your Materials and Methods: |
pKDsgRNA-FRT (LMBP 11104) is available at BCCM/GeneCorner. This plasmid was deposited by Dr T. Swings and Prof. Dr J. Michiels and was published in Swings et al., 2018. |
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.