Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V003058 | pSPV-EGFP | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pSPV-EGFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3457 bp
- Type:
- Poxvirus recombinant transfer vector
- Replication origin:
- ori
- Source/Author:
- Luo S, Peng Y, Diel DG, Rock DL.
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pSPV-EGFP 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.
pSPV-EGFP vector Sequence
LOCUS 40924_41190 3457 bp DNA circular SYN 18-DEC-2018
DEFINITION Poxvirus recombinant transfer vector pSPV-EGFP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3457)
AUTHORS Luo S, Peng Y, Diel DG, Rock DL.
TITLE Virus recombinant transfer vector pSPV-EGFP
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 3457)
AUTHORS Luo S, Peng Y, Diel DG, Rock DL.
TITLE Direct Submission
JOURNAL Submitted (05-OCT-2009) Pathobiology, University of Illinois at
Urbana-Champaign, 2001 S. Lincoln Avenue, Urbana, IL 61802, USA
REFERENCE 3 (bases 1 to 3457)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3457)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(05-OCT-2009) Pathobiology, University of Illinois at
Urbana-Champaign, 2001 S. Lincoln Avenue, Urbana, IL 61802, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3457
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 4..33
/label=multiple cloning site 1 (MCS)
/note="multiple cloning site 1 (MCS)"
regulatory 52..320
/label=early-late Vaccinia virus (VACV) VV7.5 promoter
/note="early-late Vaccinia virus (VACV) VV7.5 promoter"
/regulatory_class="promoter"
CDS 340..1056
/label=EGFP
/note="enhanced GFP"
misc_feature 1060..1085
/label=multiple cloning site 2 (MCS2)
/note="multiple cloning site 2 (MCS2)"
promoter complement(1095..1113)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(1371..1959)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2133..2990)
/label=AmpR
/note="beta-lactamase"
promoter complement(2991..3095)
/label=AmpR promoter
promoter 3441..3457
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"