Cart 2

pGL4.31[luc2P/GAL4UAS/Hygro] vector (V011519) Gene synthesis in pGL4.31[luc2P/GAL4UAS/Hygro] backbone

Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V011519 pGL4.31[luc2P/GAL4UAS/Hygro] In stock, instant shipping

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

Basic Vector Information

pGL4.31[luc2P GAL4UAS Hygro] is a vector designed for studying cell signaling using destabilized luciferase. It contains a promoter and a Gal4 upstream activating sequence, enabling the analysis of transcriptional activity in response to specific signaling pathways.

Vector Name:
pGL4.31[luc2P/GAL4UAS/Hygro]
Antibiotic Resistance:
Ampicillin
Length:
6107 bp
Type:
Luciferase Vectors
Replication origin:
ori
Source/Author:
Promega
Copy Number:
High copy number
Promoter:
SV40
Growth Strain(s):
DH10B
Growth Temperature:
37℃

pGL4.31[luc2P/GAL4UAS/Hygro] vector Map

Copy Sequence View Map
pGL4.31[luc2P/GAL4UAS/Hygro]6107 bp30060090012001500180021002400270030003300360039004200450048005100540057006000GAL4 UASAd promoterluciferasehPESTSV40 poly(A) signalSV40 promoterHygRpoly(A) signaloriAmpRsynthetic polyadenylation signalpause site

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.

pGL4.31[luc2P/GAL4UAS/Hygro] vector Sequence

LOCUS       pGL4.31[luc2P_GA        6107 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector with a promoter and a Gal4 upstream activating sequence for 
            studying cell signaling using destabilized luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    pGL4.31[luc2P GAL4UAS Hygro]
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6107)
  AUTHORS   Promega
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6107)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    31..133
                     /label=Gal4 binding site
                     /bound_moiety="Gal4"
                     /note="GAL4 UAS"
                     /note="upstream activating sequence mediating
                     Gal4-dependent induction"
     promoter        145..185
                     /note="Ad promoter"
                     /note="adenovirus major late promoter"
     CDS             238..1887
                     /label=luciferase
                     /note="firefly luciferase"
     CDS             1891..2010
                     /label=hPEST
                     /note="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
     polyA_signal    complement(2062..2183)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2377..2734
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2765..3799
                     /label=HygR
                     /note="hygromycin B phosphotransferase"
     polyA_signal    3826..3874
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4201..4789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4992..5849)
                     /label=AmpR
                     /note="beta-lactamase"
     polyA_signal    5954..6002
                     /label=synthetic polyadenylation signal
                     /note="synthetic polyadenylation signal"
     misc_feature    6016..6107
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"