ePop-EGFP vector (V014904)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014904 ePop-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

Vector Name:
ePop-EGFP
Antibiotic Resistance:
Chloramphenicol
Length:
4949 bp
Replication origin:
ori

ePop-EGFP vector Map

ePop-EGFP4949 bp6001200180024003000360042004800Lysis protein Elac operatorMyclac operator (symmetric)EGFPTranscriptional activator protein LuxRAcyl-homoserine-lactone synthaserrnB T1 terminatororilambda t0 terminatorCmRcat 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.

ePop-EGFP vector Sequence

LOCUS       V014904                 4949 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014904
VERSION     V014904
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4949)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             152..424
                     /gene="E"
                     /label="Lysis protein E"
                     /note="Lysis protein E from Enterobacteria phage phiX174.
                     Accession#: P03639"
     protein_bind    443..459
                     /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)."
     CDS             661..690
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     protein_bind    complement(867..886)
                     /label="lac operator (symmetric)"
                     /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). The
                     symmetric lac operator was optimized for tight binding of
                     lac repressor."
     CDS             953..1669
                     /label="EGFP"
                     /note="enhanced GFP"
     CDS             1706..2455
                     /gene="luxR"
                     /label="Transcriptional activator protein LuxR"
                     /note="Transcriptional activator protein LuxR from
                     Aliivibrio fischeri. Accession#: P12746"
     CDS             2495..3073
                     /gene="luxI"
                     /label="Acyl-homoserine-lactone synthase"
                     /note="Acyl-homoserine-lactone synthase from Aliivibrio
                     fischeri. Accession#: P12747"
     terminator      3122..3208
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(3373..3961)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     terminator      complement(4049..4143)
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             complement(4167..4823)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(4824..4926)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"