Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V008665 | 8xGTIIC-luciferase | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- 8xGTIIC-luciferase
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5097 bp
- Type:
- Luciferase
- Replication origin:
- ori
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- RVprimer3
- 3' Primer:
- luciferase-rev
8xGTIIC-luciferase vector Vector 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.
8xGTIIC-luciferase vector Sequence
LOCUS 40924_120 5097 bp DNA circular SYN 12-JUN-2021
DEFINITION YAP/TAZ-responsive synthetic promoter driving luciferase expression.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5097)
AUTHORS Dupont S, Morsut L, Aragona M, Enzo E, Giulitti S, Cordenonsi M,
Zanconato F, Le Digabel J, Forcato M, Bicciato S, Elvassore N,
Piccolo S
TITLE Role of YAP/TAZ in mechanotransduction.
JOURNAL Nature. 2011 Jun 8;474(7350):179-83. doi: 10.1038/nature10137.
PUBMED 21654799
REFERENCE 2 (bases 1 to 5097)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5097)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName:
"Nature"; date: "2011-06-8- 8"; volume: "474"; issue: "7350"; pages:
"179-83"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5097
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal 238..359
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS complement(403..2052)
/codon_start=1
/label=luciferase
/note="firefly luciferase"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(2193..2209)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2236..2251
/label=2xGTIIC
/bound_moiety="TEF-1 family (TEAD) proteins"
/note="dimerized GTIIC motif from the SV40 enhancer, with
high affinity for TEF-1 family (TEAD) proteins (Mahoney et
al., 2005)"
protein_bind 2268..2283
/label=2xGTIIC
/bound_moiety="TEF-1 family (TEAD) proteins"
/note="dimerized GTIIC motif from the SV40 enhancer, with
high affinity for TEF-1 family (TEAD) proteins (Mahoney et
al., 2005)"
protein_bind 2322..2337
/label=2xGTIIC
/bound_moiety="TEF-1 family (TEAD) proteins"
/note="dimerized GTIIC motif from the SV40 enhancer, with
high affinity for TEF-1 family (TEAD) proteins (Mahoney et
al., 2005)"
protein_bind 2354..2369
/label=2xGTIIC
/bound_moiety="TEF-1 family (TEAD) proteins"
/note="dimerized GTIIC motif from the SV40 enhancer, with
high affinity for TEF-1 family (TEAD) proteins (Mahoney et
al., 2005)"
misc_feature complement(2426..2517)
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
polyA_signal complement(2531..2579)
/label=poly(A) signal
/note="synthetic polyadenylation signal"
rep_origin complement(2710..3165)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3192..3296
/label=AmpR promoter
CDS 3297..4154
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4328..4916
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5070..5087
/label=L4440
/note="L4440 vector, forward primer"