pZA3PLtetO-1 luc vector (V001858) Gene synthesis in pZA3PLtetO-1 luc backbone

Price Information

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V001858 pZA3PLtetO-1 luc In stock, instant shipping

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

pZA3PLtetO-1 luc is a tetracycline-inducible plasmid containing the pLtetO-1 promoter and luciferase gene. It activates luc expression upon addition of tetracycline/Dox, enabling quantitative detection of dynamic changes in gene regulation or drug induction.

Vector Name:
pZA3PLtetO-1 luc
Antibiotic Resistance:
Chloramphenicol
Length:
3666 bp
Type:
Expression vector
Replication origin:
p15A ori
Source/Author:
Lutz R, Bujard H.
Growth Strain(s):
Stbl3

pZA3PLtetO-1 luc vector Map

pZA3PLtetO-1 luc3666 bp60012001800240030003600PLtetO-1 promoterRBSluciferaserrnB T1 terminatorp15A orilambda 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.

pZA3PLtetO-1 luc vector Sequence

LOCUS       Exported                3666 bp DNA     circular SYN 31-OCT-2025
DEFINITION  Expression vector pZA3PLtetO-1 luc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3666)
  AUTHORS   Lutz R, Bujard H.
  TITLE     Independent and tight regulation of transcriptional units in 
            Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 
            regulatory elements
  JOURNAL   Nucleic Acids Res. 25 (6), 1203-1210 (1997)
  PUBMED    9092630
REFERENCE   2  (bases 1 to 3666)
  AUTHORS   Lutz R, Bujard H.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-AUG-1996) Zentrum fuer Molekulare Biologie, University
            of Heidelberg, Im Neuenheimer Feld 282, Heidelberg 69120, Germany
REFERENCE   3  (bases 1 to 3666)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3666)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3666)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
            Acids Res."; date: "1997"; volume: "25"; issue: "6"; pages: 
            "1203-1210"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (07-AUG-1996) Zentrum fuer Molekulare Biologie, University of
            Heidelberg, Im Neuenheimer Feld 282, Heidelberg 69120, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..80
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     RBS             90..101
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             108..1757
                     /label=luciferase
                     /note="firefly luciferase"
     terminator      1776..1862
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(2032..2577)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     terminator      complement(2691..2785)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2809..3465)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3466..3568)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"