Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006697 | RN3P-KDM5B-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:
- RN3P-KDM5B-FLAG
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7915 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- T3
- Cloning Method:
- Gibson Cloning
- 5' Primer:
- M13 Reverse
- 3' Primer:
- XBG-R (GACTCCATTCGGGTGTTC)
RN3P-KDM5B-FLAG vector 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.
RN3P-KDM5B-FLAG vector Sequence
LOCUS V006697 7915 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V006697
VERSION V006697
KEYWORDS RN3P-KDM5B-FLAG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7915)
AUTHORS Zhang B, Zheng H, Huang B, Li W, Xiang Y, Peng X, Ming J, Wu X,
Zhang Y, Xu Q, Liu W, Kou X, Zhao Y, He W, Li C, Chen B, Li Y, Wang
Q, Ma J, Yin Q, Kee K, Meng A, Gao S, Xu F, Na J, Xie W
TITLE Allelic reprogramming of the histone modification H3K4me3 in early
mammalian development.
JOURNAL Nature. 2016 Sep 14;537(7621):553-557. doi: 10.1038/nature19361.
PUBMED 27626382
REFERENCE 2 (bases 1 to 7915)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7915)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName:
"Nature"; date: "2016-09-14- 14"; volume: "537"; issue: "7621";
pages: "553-557"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7915
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 432..451
/label="pBR322ori-F"
/note="pBR322 origin, forward primer"
primer_bind 685..702
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 819..840
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 855..885
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 893..909
/label="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 917..933
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 954..972
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
5'UTR 997..1039
/label="Xenopus globin 5'-UTR"
/note="translational enhancer from the 5'-UTR of the major
beta-globin gene of Xenopus laevis"
CDS 1069..1134
/label="3xFLAG"
/note="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
CDS 1180..5811
/gene="Kdm5b"
/label="Lysine-specific demethylase 5B"
/note="Lysine-specific demethylase 5B from Mus musculus.
Accession#: Q80Y84"
3'UTR 5841..5966
/label="Xenopus globin 3'-UTR"
/note="3'-UTR of the major beta-globin gene of Xenopus
laevis"
promoter complement(6057..6075)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6082..6098)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(6240..6695)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6722..6826
/label="AmpR promoter"
CDS 6827..7684
/label="AmpR"
/note="beta-lactamase"
rep_origin 7858..7915
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"