Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V002410 | pUC18T-mini-Tn7T-lux-Tp | 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
- Vector Name:
- pUC18T-mini-Tn7T-lux-Tp
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11738 bp
- Type:
- Reporter vector
- Replication origin:
- ori
- Source/Author:
- Damron FH, McKenney ES, Barbier M, Liechti GW, Schweizer HP, Goldberg JB.
- Promoter:
- Pc
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pUC18T-mini-Tn7T-lux-Tp 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.
pUC18T-mini-Tn7T-lux-Tp vector Sequence
LOCUS 40924_45078 11738 bp DNA circular SYN 18-DEC-2018
DEFINITION Reporter vector pUC18T-mini-Tn7T-lux-Tp, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11738)
AUTHORS Damron FH, McKenney ES, Barbier M, Liechti GW, Schweizer HP,
Goldberg JB.
TITLE Construction of conjugatable Mini-Tn7 Vectors for Bioluminescent
Detection and Single Copy Promoter lux Reporter Analysis in
Gram-negative Bacteria
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 11738)
AUTHORS Damron FH, McKenney ES, Barbier M, Liechti GW, Schweizer HP,
Goldberg JB.
TITLE Direct Submission
JOURNAL Submitted (30-MAR-2013) Microbiology, Immunology, and Cancer
Biology, University of Virginia, PO Box 800734, Charlottesville, VA
22908, USA
REFERENCE 3 (bases 1 to 11738)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11738)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(30-MAR-2013) Microbiology, Immunology, and Cancer Biology,
University of Virginia, PO Box 800734, Charlottesville, VA 22908,
USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..11738
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 349..365
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 393..487
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
terminator 590..676
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind 722..769
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(834..1067)
/label=TpR
/note="E. coli plasmid-associated dihydrofolate reductase"
regulatory 1263..1304
/label=promoter 2
/note="promoter 2"
/regulatory_class="promoter"
promoter complement(1393..1421)
/label=Pc promoter
/note="class 1 integron promoter"
misc_feature 1473..1520
/label=FRT
/note="FRT"
protein_bind 1473..1520
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
primer_bind 1473..1495
/label=Plux seq F
/note="Plux seq F"
promoter 1616..1644
/label=Pc promoter
/note="class 1 integron promoter"
primer_bind 2041..2063
/label=Plux seq R
/note="Plux seq R"
CDS 2491..3930
/label=LuxC
/note="LuxC fatty acid reductase"
CDS 3945..4865
/label=LuxD
/note="LuxD acyltransferase"
CDS 4917..5996
/label=LuxA
/note="LuxA luciferase subunit"
CDS 6014..6994
/label=LuxB
/note="LuxB luciferase subunit"
CDS 7176..8285
/label=LuxE
/note="LuxE"
mobile_element complement(9019..9184)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
oriT complement(9343..9451)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
rep_origin complement(9683..10271)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10445..11302)
/label=AmpR
/note="beta-lactamase"
promoter complement(11303..11407)
/label=AmpR promoter