pAAV-hSyn-eNpHR3.0-mCherry vector (V014580)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014580 pAAV-hSyn-eNpHR3.0-mCherry In stock, 1 week for quality controls

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pAAV-hSyn-eNpHR3.0-mCherry
Antibiotic Resistance:
Ampicillin
Length:
6239 bp
Type:
Gene expression
Replication origin:
ori
Host:
Mammalian cells, Adeno-associated virus
Promoter:
SYN1
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pAAV-hSyn-eNpHR3.0-mCherry vector Map

pAAV-hSyn-eNpHR3.0-mCherry6239 bp30060090012001500180021002400270030003300360039004200450048005100540057006000hSyn promoterHalorhodopsinmCherryWPREhGH poly(A) signalAAV2 ITRf1 oriAmpR promoterAmpRori

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.

pAAV-hSyn-eNpHR3.0-mCherry vector Sequence

LOCUS       V014580                 6239 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014580
VERSION     V014580
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6239)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        226..673
                     /label="hSyn promoter"
                     /note="human synapsin I promoter; confers neuron-specific
                     expression (Kugler et al., 2003)"
     CDS             713..1585
                     /gene="hop"
                     /label="Halorhodopsin"
                     /note="Halorhodopsin from Natronomonas pharaonis.
                     Accession#: P15647"
     CDS             1655..2362
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    2417..3005
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    3037..3513
                     /label="hGH poly(A) signal"
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3553..3693
                     /label="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
     rep_origin      3768..4223
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4505..4609
                     /label="AmpR promoter"
     CDS             4610..5467
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5641..6229
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"