Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007991 | pDL276 | 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:
- pDL276
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6984 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Dunny GM, Lee LN, LeBlanc DJ.
pDL276 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.
pDL276 vector Sequence
LOCUS 40924_14745 6984 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pDL276, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6984)
AUTHORS Dunny GM, Lee LN, LeBlanc DJ.
TITLE Improved electroporation and cloning vector system for gram-positive
bacteria
JOURNAL Appl. Environ. Microbiol. 57 (4), 1194-1201 (1991)
PUBMED 1905518
REFERENCE 2 (bases 1 to 6984)
AUTHORS Dunny GM, Bae T.
TITLE Direct Submission
JOURNAL Submitted (15-DEC-1999) Microbiology, University of Minnesota, Box
196 UMHC, 1460 Mayo Building, 420 Delaware St. SE, Minneapolis, MN
55455, USA
REFERENCE 3 (bases 1 to 6984)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6984)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "1991"; volume: "57"; issue: "4"; pages:
"1194-1201"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-DEC-1999) Microbiology, University of Minnesota, Box 196 UMHC,
1460 Mayo Building, 420 Delaware St. SE, Minneapolis, MN 55455, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6984
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(237..825)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 1746..2537
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
terminator 5406..5433
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 5565..5608
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator complement(5904..5931)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(6023..6109)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 6583..6599
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 6600..6656
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(6669..6685)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(6693..6709)
/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(6717..6747)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6762..6783)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."