Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007398 | pUC18T-mini-Tn7T-Zeo-gfpmut3 | In stock, 1 week for quality controls |
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-Zeo-gfpmut3
- Length:
- 5430 bp
- Type:
- Bacterial Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- EM7
pUC18T-mini-Tn7T-Zeo-gfpmut3 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-Zeo-gfpmut3 vector Sequence
LOCUS pUC18T-mini-Tn7T 5430 bp DNA circular SYN 13-MAY-2021
DEFINITION ZeoR mini-Tn7 vector, mobilizable, for GFP tagging bacteria.
ACCESSION .
VERSION .
KEYWORDS pUC18T-mini-Tn7T-Zeo-gfpmut3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5430)
AUTHORS Choi KH, Schweizer HP
TITLE mini-Tn7 insertion in bacteria with single attTn7 sites: example
Pseudomonas aeruginosa.
JOURNAL Nat Protoc. 2006;1(1):153-61.
PUBMED 17406227
REFERENCE 2 (bases 1 to 5430)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5430)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Protoc."; date: "2006"; volume: "1(1)"; pages: "153-61"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5430
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 585..601
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 629..723
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
terminator 826..912
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind complement(966..1013)
/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)."
CDS complement(1099..1470)
/label=BleoR
/note="antibiotic-binding protein"
promoter complement(1489..1536)
/label=EM7 promoter
/note="synthetic bacterial promoter"
protein_bind complement(1647..1694)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
terminator complement(1748..1834)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator complement(1937..2031)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(2054..2767)
/label=yeGFP
/note="yeast-enhanced green fluorescent protein"
RBS 2774..2785
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
mobile_element complement(2947..3112)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
oriT complement(3271..3379)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
rep_origin complement(3611..4199)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4373..5230)
/label=AmpR
/note="beta-lactamase"
promoter complement(5231..5335)
/label=AmpR promoter
primer_bind 5403..5421
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"