8xGTIIC-luciferase vector (V008665)

Basic Vector Information

      • Vector Name:
      • 8xGTIIC-luciferase
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5097 bp
      • Type:
      • Luciferase
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • RVprimer3
      • 3' Primer:
      • luciferase-rev

8xGTIIC-luciferase vector Vector Map

8xGTIIC-luciferase5097 bp6001200180024003000360042004800SV40 poly(A) signalluciferaseKS primer2xGTIIC2xGTIIC2xGTIIC2xGTIICpause sitesynthetic polyadenylation signalf1 oriAmpR promoterAmpRoriL4440

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 ℃.

8xGTIIC-luciferase vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5097 bp ds-DNA     circular SYN 12-JUN-2021
DEFINITION  YAP/TAZ-responsive synthetic promoter driving luciferase expression.
ACCESSION   .
VERSION     .
KEYWORDS    8xGTIIC-luciferase
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)
  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"
FEATURES             Location/Qualifiers
     source          1..5097
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    238..359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(275..294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     329..348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(400..2052)
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     1968..1986
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      2049..2058
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(2193..2209)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2194..2210)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     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    2426..2517
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(2458..2477)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    2531..2579
                     /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"
     primer_bind     complement(2847..2868)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3059..3078
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3192..3296
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3297..4157
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3515..3534)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4328..4916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4817..4836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5070..5087
                     /label=L4440
                     /note="L4440 vector, forward primer"

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