LOCUS       Exported               10837 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLucFXR, complete sequence.
ACCESSION   AY603762
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10837)
  AUTHORS   Houten SM, Volle DH, Cummins CL, Mangelsdorf DJ, Auwerx J.
  TITLE     In vivo imaging of farnesoid X receptor activity reveals the ileum 
            as the primary bile acid signaling tissue
  JOURNAL   Mol. Endocrinol. 21 (6), 1312-1323 (2007)
  PUBMED    17426284
REFERENCE   2  (bases 1 to 10837)
  AUTHORS   Houten SM, Auwerx J.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-APR-2004) Institut de Genetique et de Biologie 
            Moleculaire et Cellulaire, 1 rue Laurent Fries, Illkirch 67404, 
            France
REFERENCE   3  (bases 1 to 10837)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10837
                     /organism="Cloning vector pLucFXR"
                     /mol_type="other DNA"
                     /label=derived from cloning vector pGEM-7ZF(+)
                     /note="derived from cloning vector pGEM-7ZF(+)"
                     /db_xref="taxon:274547"
     primer_bind     complement(24..40)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      181..636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        714..818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             819..1679
                     /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"
     rep_origin      1850..2438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2726..2747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2762..2792
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2800..2816
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2824..2840
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2858..2876
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    2902..4124
                     /label=beta-globin insulator
                     /note="beta-globin insulator"
     misc_feature    4125..5334
                     /label=beta-globin insulator
                     /note="beta-globin insulator"
     protein_bind    5340..5504
                     /label=GAL4 binding site
                     /bound_moiety="GAL4"
     protein_bind    5385..5479
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    5510..5624
                     /label=FXR binding site
                     /bound_moiety="FXR"
     regulatory      5643..6752
                     /regulatory_class="promoter"
                     /note="Wnt-1 promoter"
     regulatory      6788..6797
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6794..8446
                     /codon_start=1
                     /product="luciferase"
                     /label=luciferase
                     /protein_id="AAT27384.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     regulatory      8447..8710
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late poly(A) signal"
     polyA_signal    8487..8608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    8722..10810
                     /label=3 flanking sequence of Wnt-1
                     /note="3 flanking sequence of Wnt-1"
     misc_feature    10817..2901
                     /label=pGEM-7ZF(+)
                     /note="pGEM-7ZF(+)"
     promoter        complement(10836..17)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"