Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017180 | pHyo094 (eMutaT7) | 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:
- pHyo094 (eMutaT7)
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 8956 bp
- Type:
- Protein expression
- Replication origin:
- p15A ori
- Host:
- E. coli
- Copy Number:
- Low
- Promoter:
- araBAD
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pHyo094 (eMutaT7) 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.
pHyo094 (eMutaT7) vector Sequence
LOCUS V017180 8956 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017180
VERSION V017180
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8956)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8956
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 43..70
/label="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 89..180
/label="AmpR promoter"
rep_origin 638..1093
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(1652..2308)
/label="CmR"
/note="chloramphenicol acetyltransferase"
promoter complement(2309..2411)
/label="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
rep_origin complement(2937..3482)
/direction=LEFT
/label="p15A ori"
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
CDS complement(3760..4635)
/label="araC"
/note="L-arabinose regulatory protein"
promoter 4662..4946
/label="araBAD promoter"
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
CDS 5009..5632
/label="PmCDA1"
/note="cytidine deaminase from the sea lamprey Petromyzon
marinus (Rogozin et al., 2007)"
CDS 5675..8320
/note="T7 RNA polymerase from Escherichia phage T7.
Accession#: P00573"
/label="T7 RNA polymerase"
CDS 8358..8609
/label="UGI"
/note="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
terminator 8821..8907
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"