pGL3 BRE Luciferase vector (V001746)

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Price Information

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V001746	pGL3 BRE Luciferase	In stock, instant shipping	Buy one, get one free!

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

Basic Vector Information

The pGL3 BRE Luciferase vector is an important tool for studying the BMP signaling pathway. It is used in BMP signaling pathway research to detect the regulatory effect of BMP on gene transcription. When transfected into cells like C2C12 and stimulated by BMP, its luciferase activity reflects the pathway's activation, being unaffected by TGF - β or activin, thus distinguishing the BMP pathway's role.
It is also used in gene transcription regulation research, helping understand how BMP regulates genes like Id1. By mutating vector elements and detecting luciferase activity, the role of each element in BMP - induced transcription activation can be determined.
The vector has high specificity for BMP signals, as its specific sequence elements allow activation only in BMP's presence and not by other factors like TGF - β, activin, or serum, enabling accurate study of the BMP pathway's role. It also has good sensitivity, as luciferase activity shows a strict dose - dependent change when cells are stimulated by different BMP concentrations, allowing precise study of the pathway's activity change.

Vector Name:: pGL3 BRE Luciferase

Antibiotic Resistance:: Ampicillin

Length:: 4942 bp

Type:: Mammalian Expression, Luciferase ; BMP/Smad Transc

Replication origin:: ori

Copy Number:: High Copy

Cloning Method:: Restriction Enzyme

5' Primer:: RVprimer3

3' Primer:: LucNRev

Growth Strain(s):: DH10B

Growth Temperature:: 37℃

pGL3 BRE Luciferase vector Map

Copy Sequence View Map

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.

References

Korchynskyi O, ten Dijke P. Identification and functional characterization of distinct critically important bone morphogenetic protein-specific response elements in the Id1 promoter. J Biol Chem. 2002 Feb 15;277(7):4883-91. doi: 10.1074/jbc.M111023200. Epub 2001 Nov 29. PMID: 11729207.

pGL3 BRE Luciferase vector Sequence

LOCUS       40924_22083        4942 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4942)
  AUTHORS   Korchynskyi O, ten Dijke P
  TITLE     Identification and functional characterization of distinct 
            critically important bone morphogenetic protein-specific response 
            elements in the Id1 promoter.
  JOURNAL   J Biol Chem. 2002 Feb 15;277(7):4883-91. Epub 2001 Nov 29.
  PUBMED    11729207
REFERENCE   2  (bases 1 to 4942)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4942)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
            Chem."; date: "2002-02-15"; volume: "277(7)"; pages: "4883-91. Epub
            2001 Nov 29"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             161..1810
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    complement(1854..1975)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2223..2240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2394..2982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3156..4013)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4014..4118)
                     /label=AmpR promoter
     rep_origin      4145..4600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    4731..4779
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    4793..4884
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"