pTRE3G-mCherry vector (V011047)

Price Information

Cat No. Plasmid Name Availability Add to cart
V011047 pTRE3G-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:
pTRE3G-mCherry
Antibiotic Resistance:
Ampicillin
Length:
4715 bp
Type:
Mammalian Expression Vectors
Replication origin:
ori
Source/Author:
Clontech
Copy Number:
High copy number
Promoter:
TRE3G

pTRE3G-mCherry vector Map

pTRE3G-mCherry4715 bp600120018002400300036004200TRE3G promotermCherryIRES2MCSsmall t intronSV40 NLSSV40 poly(A) signaloriAmpRAmpR 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.

pTRE3G-mCherry vector Sequence

LOCUS       pTRE3G-mCherry.        4715 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for doxycycline-inducible expression of a gene together with 
            the mCherry fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    pTRE3G-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4715)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4715)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..382
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     CDS             389..1096
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    1107..1694
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1701..1736
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     intron          1867..1932
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2062..2082
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    2507..2628
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2885..3473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3647..4504)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4505..4609)
                     /label=AmpR promoter