AAV-CAGGS-EGFP vector (V000429)

Price Information

Cat No. Plasmid Name Availability Add to cart
V000429 AAV-CAGGS-EGFP 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

AAV zinc finger donor vector for genomic targeting

Vector Name:
AAV-CAGGS-EGFP
Antibiotic Resistance:
Ampicillin
Length:
9809 bp
Type:
TALEN-targeted gene manipulation vectors
Replication origin:
ori
Selection Marker:
EGFP
Promoter:
CAG

AAV-CAGGS-EGFP vector Map

AAV-CAGGS-EGFP9809 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600HA-LSAT2APuroRbGH poly(A) signalCMV enhancerchicken beta-actin promoterchimeric intronEGFPbeta-globin poly(A) signalM13 revlac operatorlac promoterCAP binding siteHA-RT7 promoterM13 fwdccdBAmpRoriCAP binding sitelac promoterlac operatorM13 revT3 promoter

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.

AAV-CAGGS-EGFP vector Sequence

Copy Sequence

Download GenBank File(.gb)

LOCUS       40924_130        9809 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9809)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9809)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    33..836
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    843..868
                     /label=SA
                     /note="splice acceptor site"
     CDS             892..945
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             955..1551
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     polyA_signal    1592..1816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1822..2201
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2203..2480
                     /note="chicken beta-actin promoter"
     intron          2482..3499
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     CDS             3561..4277
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    4435..4490
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4851..4867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4875..4891)
                     /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(4899..4929)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4944..4965)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    5039..5875
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(5916..5934)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5941..5957)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6095..6394
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(7797..8654)
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8778..9366
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9654..9675
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9690..9720
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9728..9744
                     /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)."
     primer_bind     9752..9768
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9789..9807
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"