LOCUS       Exported               13271 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pDOE-15, complete sequence.
ACCESSION   KM507056
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13271)
  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 13271)
  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 13271)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13271
                     /organism="Binary vector pDOE-15"
                     /mol_type="other DNA"
                     /clone="p4453"
                     /label=FRET and subcellular localization
                     /note="FRET and subcellular localization"
                     /db_xref="taxon:1544605"
     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="AIN46660.1"
                     /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..3512
                     /codon_start=1
                     /gene="mTq2-X"
                     /product="mTurquoise2-mcs1 fusion protein"
                     /label=mTq2-X
                     /note="normalized for Arabidopsis"
                     /protein_id="AIN46661.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKGASGGGSGSMGRALGTS"
     gene            2742..3512
                     /gene="mTq2-X"
                     /label=mTq2-X
     misc_feature    2742..3458
                     /gene="mTq2-X"
                     /note="similar to protein sequence for mTurquoise2 crystal 
                     structure PDB 3ZTF; characterized in PubMed ID 22434194; 
                     mTq2, mTurquoise2 specific sequence"
     primer_bind     complement(2826..2848)
                     /label=mTq2 85R, binds mTq2 not mVenus
                     /note="mTq2 85R, binds mTq2 not mVenus"
     misc_difference 3180..3182
                     /gene="mTq2-X"
                     /label=differs from mTurquoise, mTq2=I146F
                     /note="differs from mTurquoise, mTq2=I146F"
     primer_bind     3329..3354
                     /label=mTq2 613F binds mTq2 not mTurq or mVenus
                     /note="mTq2 613F binds mTq2 not mTurq or mVenus"
     misc_feature    3459..3512
                     /label=MCS1 front end
                     /note="MCS1 front end"
     misc_feature    3513..3534
                     /label=MCS1 back end
                     /note="MCS1 back end"
     regulatory      3535..4260
                     /regulatory_class="terminator"
                     /note="OCS3octapine synthase terminator"
     terminator      3545..4252
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    3545..3553
                     /note="similar to native ocs region; c is 14nt downstream 
                     of Agro OCS stop codon (MacDonald 1991)"
     misc_feature    4261..4266
                     /label=non-unique NdeI
                     /note="non-unique NdeI"
     regulatory      4267..5600
                     /regulatory_class="promoter"
                     /note="cauliflower mosaic virus 35S promoter"
     promoter        5253..5598
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5594..5618
                     /label=pDOE mcs3bF with Bsu36I
                     /note="pDOE mcs3bF with Bsu36I"
     misc_feature    5601..5616
                     /label=Bsu361 linker
                     /note="Bsu361 linker"
     regulatory      5617..5682
                     /regulatory_class="enhancer"
                     /note="TMV-Omega enhancer"
     misc_feature    5627..5681
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     CDS             5683..6450
                     /codon_start=1
                     /gene="X-mVenus"
                     /product="mcs1-mVenus fusion protein"
                     /label=X-mVenus
                     /protein_id="AIN46662.1"
                     /translation="MGSLRSHVTSGGSGGGMVSKGEELFTGVVPILVELDGDVNGHKFS
                     VSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSA
                     MPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYN
                     SHNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQS
                     KLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     gene            5683..6450
                     /gene="X-mVenus"
                     /label=X-mVenus
     misc_feature    5683..5730
                     /gene="X-mVenus"
                     /label=MCS3 front end
                     /note="MCS3 front end"
     misc_feature    5731..6450
                     /gene="X-mVenus"
                     /note="similar to protein sequence AAZ65844.1; 
                     characterized in PubMed ID 16716067; mVenus"
     CDS             5731..6450
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_difference 6349..6351
                     /gene="X-mVenus"
                     /label=A206K monomerizing mutation
                     /note="A206K monomerizing mutation"
     misc_feature    6451..6468
                     /label=MCS3 back end
                     /note="MCS3 back end"
     regulatory      6470..6722
                     /regulatory_class="terminator"
                     /note="NOS nopaline synthase terminator"
     terminator      6470..6722
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6732..6748
                     /label=remnant from pFGC5941
                     /note="remnant from pFGC5941"
     misc_feature    6749..13271
                     /label=derived from pCAMBIA1300 via pFGC5941
                     /note="derived from pCAMBIA1300 via pFGC5941"
     primer_bind     complement(6773..6789)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   6991..7014
                     /label=right border from C58 T-DNA
                     /note="right border from C58 T-DNA"
     misc_feature    6992..7016
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      8077..9077
                     /label=pVS1
                     /note="pVS1"
     CDS             8317..8946
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             9375..10448
                     /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      10514..10708
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    11052..11192
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(11378..11966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      11481..11762
                     /label=pBR325
                     /note="pBR325"
     CDS             complement(12053..12847)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside phosphotransferase"
                     /function="kanamycin resistance"
                     /label=aadA
                     /note="AphA-3"
                     /protein_id="AIN46663.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(12053..12847)
                     /gene="aadA"
                     /label=aadA