HDAC1 Flag vector (V000945)

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V000945 HDAC1 Flag In stock, instant shipping

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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

HDAC1 Flag6869 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600CMV enhancerCMV promoterT7 promoterHistone deacetylase 1FLAGbGH poly(A) signalf1 oriSV40 promoterNeoR/KanRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR promoterpRS-marker

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

Copy Sequence

Download GenBank File(.gb)

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"