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p5xATF6 GL3 vector (V009709) Gene synthesis in p5xATF6 GL3 backbone

Price Information

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V009709 p5xATF6 GL3 In stock, instant shipping

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

Basic Vector Information

Vector Name:
p5xATF6 GL3
Antibiotic Resistance:
Ampicillin
Length:
5065 bp
Type:
Reporter vector
Replication origin:
ori
Source/Author:
Wang Y, Shen J, Arenzana N, Tirasophon W, Kaufman RJ, Prywes R.
Growth Strain(s):
Top10

p5xATF6 GL3 vector Map

Copy Sequence View Map
p5xATF6 GL35065 bp6001200180024003000360042004800c-fos minimal promoterluciferaseSV40 poly(A) signaloriAmpRAmpR promoterf1 oripoly(A) signalpause 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.

p5xATF6 GL3 vector Sequence

LOCUS       40924_2917        5065 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Reporter vector p5xATF6 GL3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5065)
  AUTHORS   Wang Y, Shen J, Arenzana N, Tirasophon W, Kaufman RJ, Prywes R.
  TITLE     Activation of ATF6 and an ATF6 DNA binding site by the endoplasmic 
            reticulum stress response
  JOURNAL   J. Biol. Chem. 275 (35), 27013-27020 (2000)
  PUBMED    10856300
REFERENCE   2  (bases 1 to 5065)
  AUTHORS   Shen J, Wang Y, Prywes R.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JUN-2005) Biological Sciences, Columbia University, 
            New York, NY 10027, USA
REFERENCE   3  (bases 1 to 5065)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5065)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Biol. 
            Chem."; date: "2000"; volume: "275"; issue: "35"; pages: 
            "27013-27020"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-JUN-2005) Biological Sciences, Columbia University, New York, NY
            10027, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      205..308
                     /label=c-fos minimal promoter
                     /note="c-fos minimal promoter"
                     /regulatory_class="promoter"
     CDS             335..1984
                     /label=luciferase
                     /note="firefly luciferase"
     polyA_signal    complement(2028..2149)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2568..3156)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3330..4187)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4188..4292)
                     /label=AmpR promoter
     rep_origin      4319..4774
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    4905..4953
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    4967..5058
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"