LOCUS       Exported               12656 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pDOE-04, complete sequence.
ACCESSION   KM507045
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12656)
  AUTHORS   Gookin TE, Assmann SM.
  TITLE     Significant reduction of BiFC non-specific assembly facilitates in 
            planta assessment of heterotrimeric G-protein interactors
  JOURNAL   Plant J. (2014) In press
  PUBMED    25187041
REFERENCE   2  (bases 1 to 12656)
  AUTHORS   Gookin TE, Assmann SM.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-SEP-2014) Department of Biology, The Pennsylvania 
            State University, 208 Mueller Laboratory, University Park, PA 16802,
            USA
REFERENCE   3  (bases 1 to 12656)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12656
                     /organism="Binary vector pDOE-04"
                     /mol_type="other DNA"
                     /clone="p4454"
                     /db_xref="taxon:1544594"
     repeat_region   1..27
                     /label=left border repeat from C58 T-DNA
                     /note="left border repeat from C58 T-DNA"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    99..104
                     /label=non-unique SacI
                     /note="non-unique SacI"
     terminator      111..363
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     regulatory      complement(111..362)
                     /regulatory_class="terminator"
                     /note="MAS mannopine synthase terminator"
     misc_feature    complement(373..406)
                     /label=BAR gene remnant
                     /note="BAR gene remnant"
     misc_feature    407..412
                     /label=MCS2 KpnI site
                     /note="MCS2 KpnI site"
     CDS             complement(413..931)
                     /codon_start=1
                     /gene="p19"
                     /product="RNA silencing suppressor 19 kDa protein"
                     /label=p19
                     /note="Tombusvirus RNA silencing suppressor 19 kDa protein;
                     similar to CAC01277.1; characterized in PubMed ID 18944038;
                     normalized for Arabidopsis"
                     /protein_id="AIN46616.1"
                     /translation="MERAIQGNDAREQANSERWDGGSGGTTSPFKLPDESPSWTEWRLH
                     NDETNSNQDNPLGFKESWGFGKVVFKRYLRYDRTEASLHRVLGSWTGDSVNYAASRFFG
                     FDQIGCTYSIRFRGVSITVSGGSRTLQHLCEMAIRSKQELLQLAPIEVESNVSRGCPEG
                     TQTFEKESE"
     gene            complement(413..931)
                     /gene="p19"
                     /label=p19
     misc_feature    932..937
                     /label=MCS2 NruI site
                     /note="MCS2 NruI site"
     regulatory      complement(935..1315)
                     /regulatory_class="promoter"
                     /note="mas1-2 mannopine synthase promoter; mas2 drives 
                     MCS2 ORF; mas1 truncated by 96 nt; enhancer elements, TATA
                     box, and start codon missing"
     promoter        complement(935..1315)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     regulatory      1328..2657
                     /regulatory_class="promoter"
                     /note="cauliflower mosaic virus 35S promoter"
     promoter        2311..2656
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2647..2671
                     /label=pDOE mcs1F with XhoI
                     /note="pDOE mcs1F with XhoI"
     misc_feature    2659..2675
                     /label=XhoSac linker
                     /note="XhoSac linker"
     misc_feature    2676..2741
                     /label=TMV omega prime translation leader
                     /note="TMV omega prime translation leader"
     misc_feature    2686..2740
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     CDS             2742..3425
                     /codon_start=1
                     /gene="NmVen210-X"
                     /product="NmVenus210-mcs1 fusion protein"
                     /label=NmVen210-X
                     /protein_id="AIN46617.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKGASGGGSGSMGR
                     ALGTS"
     gene            2742..3425
                     /gene="NmVen210-X"
                     /label=NmVen210-X
     misc_feature    2742..3371
                     /gene="NmVen210-X"
                     /label=NmVenus210 specific sequence
                     /note="NmVenus210 specific sequence"
     CDS             2742..3260
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     misc_difference 3360..3362
                     /gene="NmVen210-X"
                     /label=A206K monomerizing mutation
                     /note="A206K monomerizing mutation"
     misc_feature    3372..3425
                     /label=MCS1 front end
                     /note="MCS1 front end"
     misc_feature    3426..3447
                     /label=MCS1 back end
                     /note="MCS1 back end"
     regulatory      3448..4173
                     /regulatory_class="terminator"
                     /note="OCS3octapine synthase terminator"
     terminator      3458..4165
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    3458..3466
                     /note="similar to native ocs region; c is 14nt downstream 
                     of Agro OCS stop codon (MacDonald 1991)"
     misc_feature    4174..4179
                     /label=non-unique NdeI
                     /note="non-unique NdeI"
     regulatory      4180..5513
                     /regulatory_class="promoter"
                     /note="cauliflower mosaic virus 35S promoter"
     promoter        5166..5511
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5507..5531
                     /label=pDOE mcs3bF with Bsu36I
                     /note="pDOE mcs3bF with Bsu36I"
     misc_feature    5514..5529
                     /label=Bsu361 linker
                     /note="Bsu361 linker"
     regulatory      5530..5595
                     /regulatory_class="enhancer"
                     /note="TMV-Omega enhancer"
     misc_feature    5540..5594
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     CDS             5596..5835
                     /codon_start=1
                     /gene="CVen210-SFS-X"
                     /product="CVenus210-mcs3 fusion protein"
                     /label=CVen210-SFS-X
                     /note="StrepII-FLAG-StrepII epitopes are spatially 
                     separated"
                     /protein_id="AIN46618.1"
                     /translation="MDPNEKRDHMVLLEFVTAAGITLGMDELYKGSGSWSHPQFEKSGG
                     DYKDDDDKASWSHPQFEKSGASGMGSLRSHVTSG"
     gene            5596..5835
                     /gene="CVen210-SFS-X"
                     /label=CVen210-SFS-X
     misc_feature    5599..5685
                     /gene="CVen210-SFS-X"
                     /label=CVenus210 specific sequence
                     /note="CVenus210 specific sequence"
     misc_feature    5698..5721
                     /gene="CVen210-SFS-X"
                     /label=Strep II tag (A)
                     /note="Strep II tag (A)"
     CDS             5698..5721
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    5731..5754
                     /gene="CVen210-SFS-X"
                     /label=FLAG tag
                     /note="FLAG tag"
     CDS             5731..5754
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    5761..5784
                     /gene="CVen210-SFS-X"
                     /label=Strep II tag (B)
                     /note="Strep II tag (B)"
     CDS             5761..5784
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    5800..5835
                     /label=MCS3 front end
                     /note="MCS3 front end"
     misc_feature    5836..5853
                     /label=MCS3 back end
                     /note="MCS3 back end"
     regulatory      5855..6107
                     /regulatory_class="terminator"
                     /note="NOS nopaline synthase terminator"
     terminator      5855..6107
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6117..6133
                     /label=remnant from pFGC5941
                     /note="remnant from pFGC5941"
     misc_feature    6134..12656
                     /label=derived from pCAMBIA1300 via pFGC5941
                     /note="derived from pCAMBIA1300 via pFGC5941"
     primer_bind     complement(6158..6174)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   6376..6399
                     /label=right border from C58 T-DNA
                     /note="right border from C58 T-DNA"
     misc_feature    6377..6401
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      7462..8462
                     /label=pVS1
                     /note="pVS1"
     CDS             7702..8331
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8760..9833
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      9899..10093
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    10437..10577
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(10763..11351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      10866..11147
                     /label=pBR325
                     /note="pBR325"
     CDS             complement(11438..12232)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside phosphotransferase"
                     /function="kanamycin resistance"
                     /label=aadA
                     /note="AphA-3"
                     /protein_id="AIN46619.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(11438..12232)
                     /gene="aadA"
                     /label=aadA