LOCUS       Exported                6781 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBS 35S Rluc-attR, complete sequence.
ACCESSION   AY995136
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6781)
  AUTHORS   Subramanian C, Woo J, Cai X, Xu X, Servick S, Johnson CH, Nebenfuhr 
            A, von Arnim AG.
  TITLE     A suite of tools and application notes for in vivo protein 
            interaction assays using bioluminescence resonance energy transfer 
            (BRET)
  JOURNAL   Plant J. 48 (1), 138-152 (2006)
  PUBMED    16925598
REFERENCE   2  (bases 1 to 6781)
  AUTHORS   von Arnim AG, Subramanian C.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-MAR-2005) BCMB, University of Tennessee Knoxville, 
            1406 Circle Drive, Hesler Biology Bldg 618, Knoxville, TN 37996, USA
REFERENCE   3  (bases 1 to 6781)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6781
                     /organism="Cloning vector pBS 35S Rluc-attR"
                     /mol_type="other DNA"
                     /note="plant gateway cloning vector for bioluminescence 
                     resonance energy transfer"
                     /db_xref="taxon:324059"
     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"
     primer_bind     669..685
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      705..1459
                     /regulatory_class="promoter"
                     /note="35S promoter"
     promoter        1116..1460
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     regulatory      1460..1602
                     /regulatory_class="enhancer"
                     /note="translational enhancer; 5 UTR from tobacco etch 
                     virus"
     CDS             1610..2584
                     /codon_start=1
                     /product="renilla luciferase"
                     /label=renilla luciferase
                     /note="luciferase reporter"
                     /protein_id="AAX97732.1"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQRSTSLYKKAERET"
     CDS             1610..2542
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    2549..2673
                     /label=attR1 gateway recombination site
                     /note="attR1 gateway recombination site"
     protein_bind    2549..2673
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2698..2728
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2782..3441
                     /codon_start=1
                     /product="CmR"
                     /label=CmR
                     /note="confers resistance to antibiotic chloramphenicol"
                     /protein_id="AAX97733.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3783..4088
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /note="encodes a cytotoxic protein that is a potent poison 
                     of DNA gyrase"
                     /protein_id="AAX97734.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    4129..4253
                     /label=attR2 gateway recombination site
                     /note="attR2 gateway recombination site"
     protein_bind    complement(4129..4253)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      4324..4524
                     /regulatory_class="terminator"
                     /note="cauliflower mosaic virus 35S terminator"
     promoter        complement(4596..4614)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4635..4651)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4659..4675
                     /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(4683..4713)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4728..4749
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5037..5625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5796..6656)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="confers resistance to antibiotic ampicillin"
                     /protein_id="AAX97735.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6657..6761)
                     /gene="bla"
                     /label=AmpR promoter