HRE-luciferase vector (V007461)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V007461	HRE-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 HRE - luciferase plasmid is a crucial tool in research. It's a pGL2 vector with three hypoxia response elements (HRE) from the Pgk - 1 gene upstream of the firefly luciferase gene. This design enables it to respond to hypoxic conditions and regulate luciferase expression.
It has wide applications. In fields such as tumor research, it can be used to study the transcriptional activity of hypoxia - inducible factor 1 (HIF - 1). In gene regulation research, it reveals the roles of genes and proteins in the HIF - 1 pathway.
The plasmid is used via transfection with Mirus TransIT Transfection reagent and cotransfection with TK - Renilla luciferase for efficiency control. It has high specificity for hypoxia and good sensitivity in detecting HIF - 1 activity changes, making it valuable for research.

Vector Name:: HRE-luciferase

Antibiotic Resistance:: Ampicillin

Length:: 5820 bp

Type:: Mammalian Expression, Luciferase

Replication origin:: ori

Copy Number:: High Copy

Promoter:: Mini-TK

Cloning Method:: Restriction Enzyme

3' Primer:: Luc_N_Rev

Growth Strain(s):: DH10B

Growth Temperature:: 37℃

HRE-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

Emerling BM, Weinberg F, Liu JL, Mak TW, Chandel NS. PTEN regulates p300-dependent hypoxia-inducible factor 1 transcriptional activity through Forkhead transcription factor 3a (FOXO3a). Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):2622-7. doi: 10.1073/pnas.0706790105. Epub 2008 Feb 11. PMID: 18268343; PMCID: PMC2268186.

HRE-luciferase vector Sequence

LOCUS       40924_1339        5820 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Luciferase reporter construct containing three hypoxia response 
            elements (24-mers) from the Pgk-1 gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5820)
  AUTHORS   Emerling BM, Weinberg F, Liu JL, Mak TW, Chandel NS
  TITLE     PTEN regulates p300-dependent hypoxia-inducible factor 1 
            transcriptional activity through Forkhead transcription factor 3a 
            (FOXO3a).
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Feb 19. 105(7):2622-7.
  PUBMED    18268343
REFERENCE   2  (bases 1 to 5820)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5820)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2008 Feb 19. 105(7):2622-7."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        85..147
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             207..1856
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     intron          2100..2165
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2295..2315
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     polyA_signal    2740..2874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3016..3033)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3187..3775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3949..4806)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4807..4911)
                     /label=AmpR promoter
     rep_origin      4938..5393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    5583..5704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"