Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V008067 | pDawn | 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:
- pDawn
- Antibiotic Resistance:
- Kanamycin
- Length:
- 7213 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Ohlendorf R, Vidavski RR, Eldar A, Moffat K, Moglich A.
pDawn 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.
pDawn vector Sequence
LOCUS V008067 7213 bp DNA circular SYN 17-DEC-2018
DEFINITION Exported.
ACCESSION V008067
VERSION V008067
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7213)
AUTHORS Ohlendorf R, Vidavski RR, Eldar A, Moffat K, Moglich A.
TITLE From dusk till dawn: one-plasmid systems for light-regulated gene
expression
JOURNAL J. Mol. Biol. 416 (4), 534-542 (2012)
PUBMED 22245580
REFERENCE 2 (bases 1 to 7213)
AUTHORS Ohlendorf R, Moffat K, Moeglich A.
TITLE Direct Submission
JOURNAL Submitted (15-AUG-2011) Biophysikalische Chemie, Humboldt
Universitaet zu Berlin, Invalidenstrasse 42, Berlin 10115, Germany
REFERENCE 3 (bases 1 to 7213)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7213)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Mol.
Biol."; date: "2012"; volume: "416"; issue: "4"; pages: "534-542"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-AUG-2011) Biophysikalische Chemie, Humboldt Universitaet zu
Berlin, Invalidenstrasse 42, Berlin 10115, Germany"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7213
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(26..73)
/label="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/label="6xHis"
/note="6xHis affinity tag"
misc_feature complement(158..203)
/label="multiple cloning site"
/note="multiple cloning site"
CDS complement(205..237)
/label="T7 tag (gene 10 leader)"
/note="leader peptide from bacteriophage T7 gene 10"
CDS complement(241..258)
/label="thrombin site"
/note="thrombin recognition and cleavage site"
CDS complement(268..285)
/label="6xHis"
/note="6xHis affinity tag"
RBS complement(304..326)
/label="RBS"
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
regulatory complement(339..387)
/label="lambda pR promoter"
/note="lambda pR promoter"
/regulatory_class="promoter"
terminator complement(402..429)
/label="T7Te terminator"
/note="phage T7 early transcription terminator"
terminator complement(445..516)
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(564..596)
/label="ssrA tag (LVA)"
/note="C-terminal peptide that mediates degradation in
bacteria through the ClpXP and ClpAP proteases (McGinness
et al., 2006)"
CDS complement(597..1307)
/label="lambda repressor"
/note="phage lambda repressor"
RBS 1314..1325
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
regulatory complement(1332..1578)
/label="FixK2 promoter"
/note="FixK2 promoter"
/regulatory_class="promoter"
promoter 1873..1950
/label="lacIq promoter"
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 1951..3093
/codon_start=1
/product="YF1"
/label="YF1"
/protein_id="AFH34994.1"
/translation="MASFQSFGIPGQLEVIKKALDHVRVGVVITDPALEDNPIVYVNQG
FVQMTGYETEEILGKNCRFLQGKHTDPAEVDNIRTALQNKEPVTVQIQNYKKDGTMFWN
ELNIDPMEIEDKTYFVGIQNDITEHQQTQARLQELQSELVHVSRLSAMGEMASALAHEL
NQPLAAISNYMKGSRRLLAGSSDPNTPKVESALDRAAEQALRAGQIIRRLRDFVARGES
EKRVESLSKLIEEAGALGLAGAREQNVQLRFSLDPGADLVLADRVQIQQVLVNLFRNAL
EAMAQSQRRELVVTNTPAADDMIEVEVSDTGSGFQDDVIPNLFQTFFTTKDTGMGVGLS
ISRSIIEAHGGRMWAESNASGGATFRFTLPAADEMIGGLA"
CDS 3090..3704
/gene="fixJ"
/label="Transcriptional regulatory protein FixJ"
/note="Transcriptional regulatory protein FixJ from
Bradyrhizobium diazoefficiens (strain JCM 10833 / BCRC
13528 / IAM 13628 / NBRC 14792 / USDA 110). Accession#:
P23221"
CDS 4508..4696
/label="rop"
/note="Rop protein, which maintains plasmids at low copy
number"
misc_feature 4801..4943
/label="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(5129..5717)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 5839..6651
/label="KanR"
/note="aminoglycoside phosphotransferase"
rep_origin complement(6747..7202)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"