Basic Vector Information
- 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
HRE-luciferase vector Vector Map
Plasmid Resuspension 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 ℃.
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"
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