Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001739 | pDL278 | 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:
- pDL278
- Antibiotic Resistance:
- Spectinomycin
- Length:
- 6733 bp
- Type:
- Bacterial Expression ; E. coli/Gram-positive Bacte
- Replication origin:
- ori
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- M13_pUC_rev
- 3' Primer:
- M13_pUC_fwd; pBAD_rev
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pDL278 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.
pDL278 vector Sequence
LOCUS Exported 6733 bp DNA circular SYN 16-OCT-2024
DEFINITION E. Coli/Gram-positive Bacterial Shuttle Vector.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6733)
AUTHORS LeBlanc DJ, Lee LN, Abu-Al-Jaibat A
TITLE Molecular, genetic, and functional analysis of the basic replicon of
pVA380-1, a plasmid of oral streptococcal origin.
JOURNAL Plasmid. 1992 Sep;28(2):130-45.
PUBMED 1409970
REFERENCE 2 (bases 1 to 6733)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6733)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6733)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid.";
date: "1992-09"; volume: "28(2)"; pages: "130-45"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6733
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(66..83)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(237..825)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 1775..2449
/codon_start=1
/label=Spec
/note="spectinomycin adenyltransferase"
/translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
KL"
CDS complement(2705..3670)
/codon_start=1
/label=Rep
/translation="MTGTENQVFKDYSKNGKDRKWRERKLKNIELAGRLERLGYRSFER
VYQCAEVLKFIEQQDGTKKLYQSYFCKNKLCPICNWRRSMKYAYQAELVVNEAMKRYPK
GRFLFLTLTIKNISGEKLNKSISEIGRAFNRLMKYKKVDKNVIGYLRATEVTYSTEHEN
YHPHLHVLLFVKSSYFTGNNTNYISQEEWTKLWAKAMKLDYTPVVDIRTVKAHKRKNLK
SAIIETAKYPVKPFDVDTEDVTLFSEMVKERITEDLTNGLHRKRQIGFGKLFKKIKAEL
ALDDVEEGNLVQTGAEESAESTGREIVAFWNWDRKNYFVR"
terminator 5155..5182
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 5314..5357
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator complement(5653..5680)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(5772..5858)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 6332..6348
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 6349..6405
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(6418..6434)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(6442..6458)
/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(6466..6496)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6511..6532)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."