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PB-TRE3G-SOX17 vector (V014467)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014467 PB-TRE3G-SOX17 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:
PB-TRE3G-SOX17
Antibiotic Resistance:
Ampicillin
Length:
6425 bp
Type:
Protein expression, Tetracycline regulation
Replication origin:
ori
Host:
Mammalian cells
Promoter:
TRE3G
Growth Temperature:
37℃

PB-TRE3G-SOX17 vector Map

Copy Sequence View Map
PB-TRE3G-SOX176425 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300M13 fwdTRE3G promoterattB1Transcription factor SOX-17attB2SV40 poly(A) signallox2272M13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR 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.

PB-TRE3G-SOX17 vector Sequence

LOCUS       V014467                 6425 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014467
VERSION     V014467
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6425)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     381..397
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        1430..1808
                     /label="TRE3G promoter"
                     /note="3rd-generation Tet-responsive promoter that can be
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1826..1850
                     /label="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1868..3109
                     /gene="SOX17"
                     /label="Transcription factor SOX-17"
                     /note="Transcription factor SOX-17 from Homo sapiens.
                     Accession#: Q9H6I2"
     protein_bind    complement(3129..3153)
                     /label="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    complement(3203..3324)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3795..3828)
                     /label="lox2272"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are
                     compatible with each other, but incompatible with loxP or
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(4206..4222)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4230..4246)
                     /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(4254..4284)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4299..4320)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4608..5196)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5370..6227)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6228..6332)
                     /label="AmpR promoter"