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"