pGEN-luxCDABE-Chl vector (V014593)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014593 pGEN-luxCDABE-Chl 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:
pGEN-luxCDABE-Chl
Antibiotic Resistance:
Chloramphenicol
Length:
11113 bp
Type:
Gene expression
Replication origin:
RSF ori
Host:
E. coli
Promoter:
EM7
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pGEN-luxCDABE-Chl vector Map

pGEN-luxCDABE-Chl11113 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000EM7 promoterLuxCLuxDLuxALuxBLuxERSF orirrnB T1 terminatorCmRcat promoterPlasmid segregation protein ParM

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.

pGEN-luxCDABE-Chl vector Sequence

LOCUS       V014593                11113 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014593
VERSION     V014593
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11113)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        887..934
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             1021..2460
                     /label="LuxC"
                     /note="LuxC fatty acid reductase"
     CDS             2476..3396
                     /label="LuxD"
                     /note="LuxD acyltransferase"
     CDS             3448..4527
                     /label="LuxA"
                     /note="LuxA luciferase subunit"
     CDS             4545..5525
                     /label="LuxB"
                     /note="LuxB luciferase subunit"
     CDS             5707..6816
                     /label="LuxE"
                     /note="LuxE"
     rep_origin      complement(7074..7823)
                     /direction=LEFT
                     /label="RSF ori"
                     /note="Plasmids containing the RSF 1030 origin of
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     terminator      complement(7987..8073)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(8791..9447)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(9448..9550)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             join(10470..11113,1..268)
                     /gene="parM"
                     /label="Plasmid segregation protein ParM"
                     /note="Plasmid segregation protein ParM from Escherichia
                     coli. Accession#: P11904"