pEnCAG-3×HA-SIRT5 vector (V014009)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V014009	pEnCAG-3×HA-SIRT5	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:: pEnCAG-3×HA-SIRT5

Antibiotic Resistance:: Ampicillin

Length:: 8093 bp

Type:: Protein expression

Replication origin:: ori

Host:: Mammalian cells

Selection Marker:: Neo/G418

Promoter:: CAG

Growth Strain(s):: DH5a

Growth Temperature:: 37℃

pEnCAG-3×HA-SIRT5 vector Map

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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.

pEnCAG-3×HA-SIRT5 vector Sequence

LOCUS       V014009                 8093 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014009
VERSION     V014009
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8093)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          709..1726
                     /label="chimeric intron"
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     CDS             1842..1868
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             1872..1898
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             1902..1928
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             1929..2858
                     /gene="SIRT5"
                     /label="NAD-dependent protein deacylase sirtuin-5,
                     mitochondrial"
                     /note="NAD-dependent protein deacylase sirtuin-5,
                     mitochondrial from Homo sapiens. Accession#: Q9NXA8"
     misc_feature    2869..3457
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    3523..3571
                     /label="HSV TK poly(A) signal"
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3773..4201
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4215..4544
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             4611..5402
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5581..5714
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5751..5767)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5775..5791)
                     /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(5799..5829)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5844..5865)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6153..6741)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6915..7772)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7773..7877)
                     /label="AmpR promoter"