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pTet-LasR-PluxR-lacZ vector (V013852)

Price Information

Cat No. Plasmid Name Availability Add to cart
V013852 pTet-LasR-PluxR-lacZ 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:
pTet-LasR-PluxR-lacZ
Antibiotic Resistance:
Ampicillin
Length:
6979 bp
Type:
Protein expression
Replication origin:
ori
Host:
E. coli
Promoter:
PluxR
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pTet-LasR-PluxR-lacZ vector Map

Copy Sequence View Map
pTet-LasR-PluxR-lacZ6979 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900M13 fwdtet operatorTranscriptional activator protein LasRrrnB T1 terminatorT7Te terminatorlacZrrnB T1 terminatorM13 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.

pTet-LasR-PluxR-lacZ vector Sequence

LOCUS       V013852                 6979 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013852
VERSION     V013852
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6979)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    405..423
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             490..1206
                     /gene="lasR"
                     /label="Transcriptional activator protein LasR"
                     /note="Transcriptional activator protein LasR from
                     Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP
                     104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1).
                     Accession#: P25084"
     terminator      1221..1292
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1308..1335
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     CDS             1551..4595
                     /label="lacZ"
                     /note="beta-galactosidase"
     terminator      complement(4616..4659)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(4758..4774)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4782..4798)
                     /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(4806..4836)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4851..4872)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5160..5748)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5922..6779)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6780..6884)
                     /label="AmpR promoter"