Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010494 | pENTR4-FLAG | 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:
- pENTR4-FLAG
- Antibiotic Resistance:
- Kanamycin
- Length:
- 3401 bp
- Type:
- Gateway vectors
- Replication origin:
- ori
- Copy Number:
- Low copy number
- Cloning Method:
- Enzyme digestion and ligation
- 5' Primer:
- ENTR-F: CTACAAACTCTTCCTGTTAGTTAG
- 3' Primer:
- ENTR-R: ATGGCTCATAACACCCCTTG
- Fusion Tag:
- N-FLAG
pENTR4-FLAG 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.
pENTR4-FLAG vector Sequence
LOCUS 40924_17532 3401 bp DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3401)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 3401)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3401
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 103..189
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 358..457
/label=attL1
/note="recombination site for the Gateway(R) LR reaction"
CDS 466..489
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
protein_bind complement(562..661)
/gene="mutant version of attL"
/label=LR Clonase(TM) binding site
/bound_moiety="LR Clonase(TM)"
/note="attL2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 784..1590
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 1683..2271
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator 2562..2605
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2737..2764
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 2828..2927
/gene="mutant version of attL"
/label=LR Clonase(TM) binding site
/bound_moiety="LR Clonase(TM)"
/note="attL1"
/note="recombination site for the Gateway(R) LR reaction"
CDS 2936..2959
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
protein_bind complement(3032..3131)
/label=attL2
/note="recombination site for the Gateway(R) LR reaction"