Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006439 | pEX18Gm | 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
pEX18GM is a SacB suicide plasmid of Escherichia coli. Please note: the full-length sequencing results showed several mutations aligned with the theoretical sequence, but none of them in the functional regions.
- Vector Name:
- pEX18Gm
- Antibiotic Resistance:
- Gentamicin
- Length:
- 5831 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP.
- Promoter:
- sacB
- Growth Strain(s):
- DH5alpha
- Growth Temperature:
- 37℃
pEX18Gm vector 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
- Guo R, Li G, Lu L, Sun S, Liu T, Li M, Zheng Y, Walhout AJM, Wu J, Li H. The Plasmid pEX18Gm Indirectly Increases Caenorhabditis elegans Fecundity by Accelerating Bacterial Methionine Synthesis. Int J Mol Sci. 2022 Apr 30;23(9):5003.
pEX18Gm vector Sequence
LOCUS 40924_18756 5831 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pEX18Gm, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5831)
AUTHORS Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP.
TITLE A broad-host-range Flp-FRT recombination system for site-specific
excision of chromosomally-located DNA sequences: application for
isolation of unmarked Pseudomonas aeruginosa mutants
JOURNAL Gene 212 (1), 77-86 (1998)
PUBMED 9661666
REFERENCE 2 (bases 1 to 5831)
AUTHORS Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (10-FEB-1998) Microbiology, Colorado State University,
Fort Collins, CO 80523, USA
REFERENCE 3 (bases 1 to 5831)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5831)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene";
date: "1998"; volume: "212"; issue: "1"; pages: "77-86"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(10-FEB-1998) Microbiology, Colorado State University, Fort Collins,
CO 80523, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5831
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(212..1630)
/label=SacB
/note="secreted levansucrase that renders bacterial growth
sensitive to sucrose"
promoter complement(1631..2076)
/label=sacB promoter
/note="sacB promoter and control region"
oriT complement(2450..2559)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
terminator complement(2980..3007)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(3099..3185)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rRNA complement(3186..3305)
/product="Eschericia coli 5S ribosomal RNA"
primer_bind 3535..3551
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(3555..3611)
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(3621..3637)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3645..3661)
/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)."
promoter complement(3669..3699)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3714..3735)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4023..4611)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4867..5397)
/label=GmR
/note="gentamycin acetyltransferase"
promoter complement(5586..5614)
/label=Pc promoter
/note="class 1 integron promoter"
promoter complement(5632..5736)
/label=AmpR promoter