Cart 2

vYM112_PB-pEF1a-optiPIN2-HA(Hygro) vector (V014495)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014495 vYM112_PB-pEF1a-optiPIN2-HA(Hygro) 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:
vYM112_PB-pEF1a-optiPIN2-HA(Hygro)
Antibiotic Resistance:
Ampicillin
Length:
9590 bp
Type:
Protein expression, Transposition
Replication origin:
ori
Host:
Mammalian cells
Selection Marker:
Hyg
Promoter:
EF-1α
Growth Temperature:
30℃

vYM112_PB-pEF1a-optiPIN2-HA(Hygro) vector Map

Copy Sequence View Map
vYM112_PB-pEF1a-optiPIN2-HA(Hygro)9590 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200oriAmpRAmpR promoterf1 oriM13 fwdEF-1-alpha promoterAuxin efflux carrier component 2HAbGH poly(A) signalSV40 promoterHygRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding site

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.

vYM112_PB-pEF1a-optiPIN2-HA(Hygro) vector Sequence

LOCUS       V014495                 9590 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014495
VERSION     V014495
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9590)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(230..818)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(992..1849)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1850..1954)
                     /label="AmpR promoter"
     rep_origin      complement(1980..2435)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2577..2593
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3228..4406
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             4465..6405
                     /gene="PIN2"
                     /label="Auxin efflux carrier component 2"
                     /note="Auxin efflux carrier component 2 from Arabidopsis
                     thaliana. Accession#: Q9LU77"
     CDS             6418..6444
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     polyA_signal    6478..6685
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        6723..7052
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             7101..8123
                     /label="HygR"
                     /note="aminoglycoside phosphotransferase from E. coli"
     polyA_signal    8249..8370
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9418..9434)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(9442..9458)
                     /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(9466..9496)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(9511..9532)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."