LOCUS       Exported               12483 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pPZP 35S hRluc-attR, complete sequence.
ACCESSION   AY995142
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12483)
  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 12483)
  AUTHORS   von Arnim AG, Subramanian C.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2005) BCMB, University of Tennessee Knoxville, 
            1406 Circle Drive, Hesler Biology Bldg 618, Knoxville, TN 37996, USA
REFERENCE   3  (bases 1 to 12483)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12483
                     /organism="Binary vector pPZP 35S hRluc-attR"
                     /mol_type="other DNA"
                     /note="plant gateway cloning vector for bioluminescence 
                     resonance energy transfer"
                     /db_xref="taxon:324065"
     CDS             1187..1816
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2250..3317
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      3383..3577
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    3921..4061
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4247..4835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5079..6011)
                     /codon_start=1
                     /product="spectinomycin adenyltransferase"
                     /label=spectinomycin adenyltransferase
                     /note="confers resistance to antibiotic spectinomycin"
                     /protein_id="AAX97756.1"
                     /translation="MTCFFGVQSMPRASKQQARYAVGRCLMLWSSNDVTQQGSRPKTKL
                     NIMGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVR
                     LDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRND
                     ILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNS
                     PPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQ
                     EEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(5079..5870)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6397..6422
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    6398..6422
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    6520..6694
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     6909..6925
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      6946..7700
                     /regulatory_class="promoter"
                     /note="35S cauliflower mosaic virus promoter"
     promoter        7357..7701
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             7851..8825
                     /codon_start=1
                     /product="renilla luciferase"
                     /label=renilla luciferase
                     /note="human codon optimized renilla luciferase reporter"
                     /protein_id="AAX97757.1"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQRSTSLYKKAERET"
     misc_feature    8790..8914
                     /label=attR1 gateway recombination site
                     /note="attR1 gateway recombination site"
     protein_bind    8790..8914
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        8939..8969
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             9023..9682
                     /codon_start=1
                     /product="chloramphenicol acetyl transferase"
                     /label=chloramphenicol acetyl transferase
                     /note="confers resistance to antibiotic chloramphenicol"
                     /protein_id="AAX97758.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             10024..10329
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /note="encodes a cytotoxic protein that is a potent poison 
                     of DNA gyrase"
                     /protein_id="AAX97759.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    10370..10494
                     /label=attR2 gateway recombination site
                     /note="attR2 gateway recombination site"
     protein_bind    complement(10370..10494)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      10565..10775
                     /regulatory_class="terminator"
                     /note="35S cauliflower mosaic virus terminator"
     promoter        10840..11220
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             11226..11777
                     /codon_start=1
                     /product="phosphinothricin acetyl transferase"
                     /label=phosphinothricin acetyl transferase
                     /note="confers resistance to herbicide BASTA"
                     /protein_id="AAX97760.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      11787..12039
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     primer_bind     complement(12067..12083)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    12091..12107
                     /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(12115..12145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12160..12181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    complement(12346..12371)
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    12346..12370
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"