Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V003835 | pRAB11 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
Plasmid pRAB11 was generated from pRMC2 by adding a second tet operator within the TetR-regulated promoter Pxyl/tet. Pronounced repression was observed in the absence of anhydrotetracycline (ATc) combined with high induction in the presence of the drug, as demonstrated for pRAB11 bearing staphylococcal nuclease nuc1, lacZ or gfp.
- Vector Name:
- pRAB11
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6447 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Helle L, Kull M, Mayer S, Marincola G, Zelder ME, Goerke C, Wolz C, Bertram R.
- Promoter:
- pUC
- Growth Strain(s):
- DH10b
- Growth Temperature:
- 37℃
pRAB11 vector Map
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5. Store the plasmid at -20 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- Wu Y, Meng Y, Qian L, Ding B, Han H, Chen H, Bai L, Qu D, Wu Y. The Vancomycin Resistance-Associated Regulatory System VraSR Modulates Biofilm Formation of Staphylococcus epidermidis in an ica-Dependent Manner. mSphere. 2021 Oct 27;6(5):e0064121. doi: 10.1128/mSphere.00641-21. Epub 2021 Sep 22. PMID: 34550006; PMCID: PMC8550092.
pRAB11 vector Sequence
LOCUS V003835 6447 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V003835
VERSION V003835
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6447)
AUTHORS Helle L, Kull M, Mayer S, Marincola G, Zelder ME, Goerke C, Wolz C,
Bertram R.
TITLE Vectors for improved Tet repressor-dependent gradual gene induction
or silencing in Staphylococcus aureus
JOURNAL Microbiology (Reading, Engl.) 157 (PT 12), 3314-3323 (2011)
PUBMED 21921101
REFERENCE 2 (bases 1 to 6447)
AUTHORS Bertram R.
TITLE Direct Submission
JOURNAL Submitted (30-AUG-2011) Dept of Microbial Genetics, University of
Tuebingen, Waldhaeuser Str. 70/8, Tuebingen 72076, Germany
REFERENCE 3 (bases 1 to 6447)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6447)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Microbiology (Reading, Engl.) 157 (PT 12), 3314-3323 (2011)"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(30-AUG-2011) Dept of Microbial Genetics, University of Tuebingen,
Waldhaeuser Str. 70/8, Tuebingen 72076, Germany"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6447
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 244..265
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 280..310
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 318..334
/label="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 342..358
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(904..1551)
/gene="cat"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
rep_origin 2009..2944
CDS 3042..3056
/label="enterokinase site"
/note="enterokinase recognition and cleavage site"
CDS complement(3371..3994)
/label="TetR"
/note="tetracycline repressor TetR"
regulatory complement(4010..4065)
/label="pR* w/o tetO"
/note="pR* w/o tetO"
/regulatory_class="promoter"
protein_bind 4101..4119
/label="tet operator"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 4136..4153
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
primer_bind complement(4189..4205)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 4679..4783
/label="AmpR promoter"
CDS 4784..5641
/label="AmpR"
/note="beta-lactamase"
rep_origin 5815..6403
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"