Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V012503 | pF3A WG (BYDV) (SSA) | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pF3A WG (BYDV) (SSA)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3689 bp
- Type:
- Cloning Vectors
- Replication origin:
- ori
- Source/Author:
- Promega
- Copy Number:
- High copy number
- Promoter:
- SP6
- Growth Strain(s):
- Stbl3
pF3A WG (BYDV) (SSA) 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.
pF3A WG (BYDV) (SSA) vector Sequence
LOCUS pF3A_WG_(BYDV). 3689 bp DNA circular SYN 01-JAN-1980
DEFINITION Flexi(R) vector with an ampicillin resistance marker, for cell-free
translation in a wheat germ extract.
ACCESSION .
VERSION .
KEYWORDS pF3A WG (BYDV)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3689)
AUTHORS Promega
TITLE Direct Submission
REFERENCE 2 (bases 1 to 3689)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Subclone a coding sequence between the SgfI/AsiSI and PmeI sites to
remove the lethal barnase gene.
FEATURES Location/Qualifiers
source 1..3689
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 23..41
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 46..63
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
5'UTR 65..201
/label=BYDV 5' UTR
/note="barley yellow dwarf virus 5' untranslated region;
required for function of the cap-independent 3' translation
enhancer (Guo et al., 2001)"
CDS 233..565
/label=barnase
/note="ribonuclease from Bacillus amyloliquefaciens"
misc_feature 596..702
/label=BYDV 3' TE
/note="cap-independent 3' translation enhancer from barley
yellow dwarf virus (Wang et al., 1997)"
terminator 807..854
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 1083..1187
/label=AmpR promoter
CDS 1188..2045
/label=AmpR
/note="beta-lactamase"
rep_origin complement(2206..2794)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2910..3193
/label=cer region
/note="ColE1-derived recombination site that helps to
maintain plasmids as monomers"
terminator 3423..3509
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3601..3628
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"