Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017305 | pSecUAG-Evol2 | 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:
- pSecUAG-Evol2
- Antibiotic Resistance:
- Kanamycin
- Length:
- 8649 bp
- Type:
- Protein expression
- Replication origin:
- RSF ori
- Host:
- E. coli
- Copy Number:
- Low
- Promoter:
- araBAD
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pSecUAG-Evol2 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.
pSecUAG-Evol2 vector Sequence
LOCUS V017305 8649 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017305
VERSION V017305
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8649)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8649
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(1..285)
/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 312..1187
/label="araC"
/note="L-arabinose regulatory protein"
misc_feature 1454..1594
/label="bom"
/note="basis of mobility region from pBR322"
rep_origin 1815..2563
/label="RSF ori"
/note="Plasmids containing the RSF 1030 origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
promoter 2579..2670
/label="AmpR promoter"
CDS 2671..3483
/label="KanR"
/note="aminoglycoside phosphotransferase"
CDS 3514..4731
/gene="sufS"
/label="Cysteine desulfurase"
/note="Cysteine desulfurase from Escherichia coli (strain
K12). Accession#: P77444"
promoter 4741..4788
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 6461..7495
/gene="selD"
/label="Selenide, water dikinase"
/note="Selenide, water dikinase from Aeromonas salmonicida
(strain A449). Accession#: A4SMM1"
terminator complement(8214..8300)
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"