Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017257 | pHyo245 | 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:
- pHyo245
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10178 bp
- Type:
- Gene expression
- Replication origin:
- ori2
- Host:
- E. coli
- Copy Number:
- Low
- Promoter:
- T7
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pHyo245 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.
pHyo245 vector Sequence
LOCUS V017257 10178 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017257
VERSION V017257
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10178)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..10178
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(820..867)
/label="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 915..933
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS complement(966..983)
/label="6xHis"
/note="6xHis affinity tag"
CDS complement(993..1973)
/gene="pheS"
/label="Phenylalanine--tRNA ligase alpha subunit"
/note="Phenylalanine--tRNA ligase alpha subunit from
Escherichia coli O139:H28 (strain E24377A / ETEC).
Accession#: A7ZMI2"
CDS complement(1977..1994)
/label="thrombin site"
/note="thrombin recognition and cleavage site"
CDS complement(2004..2021)
/label="6xHis"
/note="6xHis affinity tag"
RBS complement(2040..2062)
/label="RBS"
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind complement(2077..2101)
/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(2102..2120)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
terminator 2198..2245
/label="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 2477..2554
/label="lacI promoter"
CDS 2555..3634
/label="lacI"
/note="lac repressor"
protein_bind 3650..3671
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(3957..4814)
/label="AmpR"
/note="beta-lactamase"
promoter complement(4815..4906)
/label="AmpR promoter"
rep_origin complement(5473..6186)
/direction=LEFT
/label="oriV"
/note="incP origin of replication"
rep_origin 6265..6484
/label="ori2"
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
CDS 6575..7327
/label="repE"
/note="replication initiation protein for the bacterial F
plasmid"
misc_feature 7333..7583
/label="incC"
/note="incompatibility region of the bacterial F plasmid"
CDS 7909..9081
/label="sopA"
/note="partitioning protein for the bacterial F plasmid"
CDS 9084..10052
/label="sopB"
/note="partitioning protein for the bacterial F plasmid"