Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V000945 | HDAC1 Flag | 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:
- HDAC1 Flag
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6869 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- SV40
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- T7
- 3' Primer:
- BGH Reverse
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
HDAC1 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.
HDAC1 Flag vector Sequence
LOCUS V000945 6869 bp DNA circular SYN 13-MAY-2021 DEFINITION Exported. ACCESSION V000945 VERSION V000945 KEYWORDS HDAC1 Flag SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 6869) AUTHORS Emiliani S, Fischle W, Van Lint C, Al-Abed Y, Verdin E TITLE Characterization of a human RPD3 ortholog, HDAC3. JOURNAL Proc Natl Acad Sci U S A. 1998 Mar 17. 95(6):2795-800. PUBMED 9501169 REFERENCE 2 (bases 1 to 6869) TITLE Direct Submission REFERENCE 3 (bases 1 to 6869) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl Acad Sci U S A. 1998 Mar 17. 95(6):2795-800." SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..6869 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 74..453 /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 454..657 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" promoter 702..720 /label="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 783..2228 /gene="HDAC1" /label="Histone deacetylase 1" /note="Histone deacetylase 1 from Homo sapiens. Accession#: Q13547" CDS 2241..2264 /label="FLAG" /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" polyA_signal 2305..2529 /label="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" rep_origin 2575..3003 /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3017..3346 /label="SV40 promoter" /note="SV40 enhancer and early promoter" CDS 3413..4204 /label="NeoR/KanR" /note="aminoglycoside phosphotransferase" polyA_signal 4381..4514 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" primer_bind complement(4551..4567) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind complement(4575..4591) /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)." promoter complement(4599..4629) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(4644..4665) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4782..4799) /label="L4440" /note="L4440 vector, forward primer" rep_origin complement(4953..5541) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5715..6572) /label="AmpR" /note="beta-lactamase" promoter complement(6573..6677) /label="AmpR promoter" primer_bind complement(6752..6771) /label="pRS-marker" /note="pRS vectors, use to sequence yeast selectable marker"