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pAV-U6+27-Tornado-F30-TAR Variant-1 vector (V015622)

Price Information

Cat No. Plasmid Name Availability Add to cart
V015622 pAV-U6+27-Tornado-F30-TAR Variant-1 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:
pAV-U6+27-Tornado-F30-TAR Variant-1
Antibiotic Resistance:
Ampicillin
Length:
6243 bp
Type:
Transcriptional regulation
Replication origin:
ori
Host:
Mammalian cells, Adeno-associated virus
Selection Marker:
Neo/G418
Promoter:
U6
Growth Temperature:
37℃

pAV-U6+27-Tornado-F30-TAR Variant-1 vector Map

Copy Sequence View Map
pAV-U6+27-Tornado-F30-TAR Variant-16243 bp30060090012001500180021002400270030003300360039004200450048005100540057006000SV40 promoterNeoR/KanRsmall t intronSV40 NLSSV40 poly(A) signaloriAmpRAmpR promoterf1 oriSV40 poly(A) signalU6 promoterSV40

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.

pAV-U6+27-Tornado-F30-TAR Variant-1 vector Sequence

LOCUS       62056_3240        6243 bp DNA     circular SYN 10-SEP-2019
DEFINITION  Cloning vector pAV-U6+27-Tornado-F30-TAR Variant-1, complete 
            sequence.
ACCESSION   MN052909
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6243)
  AUTHORS   Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR.
  TITLE     Live imaging of mRNA using RNA-stabilized fluorogenic proteins
  JOURNAL   Nat. Methods 16 (9), 862-865 (2019)
  PUBMED    31471614
REFERENCE   2  (bases 1 to 6243)
  AUTHORS   Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUN-2019) Pharmacology, Weill Cornell Medicine, 1300 
            York Avenue, New York, NY 10022, United States
REFERENCE   3  (bases 1 to 6243)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2019"; volume: "16"; issue: "9"; pages: "862-865"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-JUN-2019) Pharmacology, Weill Cornell Medicine, 1300 York 
            Avenue, New York, NY 10022, United States"
FEATURES             Location/Qualifiers
     source          1..6243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     misc_RNA        308..582
                     /product="Tornado-F30-TAR Variant-1"
     CDS             689..1480
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     intron          1903..1968
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2098..2118
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    2543..2677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2946..3534)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3708..4565)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4566..4670)
                     /label=AmpR promoter
     rep_origin      4697..5152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    complement(5424..5545)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(5973..6213)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     promoter        6237..6243
                     /label=SV40
                     /note="Simian virus 40 enhancer/early promoter"