LOCUS       Exported                5051 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pBS-35S-Ala-Rluc, complete sequence.
ACCESSION   AY189982
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5051)
  AUTHORS   von Arnim A.
  TITLE     Vectors for Bioluminescence Resonance Energy Transfer
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5051)
  AUTHORS   von Arnim A.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-DEC-2002) Botany, University of Tennessee, 1406 Circle
            Drive, Knoxville, TN 37996, USA
REFERENCE   3  (bases 1 to 5051)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5051
                     /organism="Expression vector pBS-35S-Ala-Rluc"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:221061"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    653..704
                     /label=polylinker
                     /note="polylinker"
     primer_bind     669..685
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      705..1459
                     /regulatory_class="promoter"
                     /note="cauliflower mosaic virus dual 35S promoter"
     promoter        1116..1460
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     5UTR           1460..1602
                     /note="translational enhancer; 5UTR from tobacco etch 
                     virus"
     regulatory      1601..1610
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1603..2589
                     /codon_start=1
                     /gene="Rluc-AlaN"
                     /product="renilla luciferase"
                     /label=Rluc-AlaN
                     /note="luciferase reporter; contains a N-terminal alanine 
                     linker"
                     /protein_id="AAO48593.1"
                     /translation="MAAAAPVAAAAAALMASKVYDPEQRKRMITGPQWWARCKQMNVLD
                     SFINYYDSEKHAENAVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNG
                     SYRLLDHYKYLTAWFELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDV
                     IESWDEWPDIEEDIALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKE
                     KGEVRRPTLSWPREIPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIV
                     EGAKKFPNTEFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQRSI"
     gene            1603..2589
                     /gene="Rluc-AlaN"
                     /label=Rluc-AlaN
     CDS             1645..2577
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     regulatory      2590..2800
                     /regulatory_class="terminator"
                     /note="cauliflower mosaic virus 35S terminator"
     promoter        complement(2866..2884)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2905..2921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2929..2945
                     /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)."
     promoter        complement(2953..2983)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2998..3019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3307..3895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4066..4926)
                     /codon_start=1
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=beta-lactamase
                     /protein_id="AAO48594.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4927..5031)
                     /gene="bla"
                     /label=AmpR promoter