LOCUS       Exported               11919 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pDOE-18, complete sequence.
ACCESSION   KM507059
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11919)
  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 11919)
  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 11919)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11919
                     /organism="Binary vector pDOE-18"
                     /mol_type="other DNA"
                     /clone="p4577"
                     /label=FRET and subcellular localization
                     /note="FRET and subcellular localization"
                     /db_xref="taxon:1544608"
     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="AIN46672.1"
                     /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..1965
                     /regulatory_class="promoter"
                     /note="UBQ10; Arabidopsis ubiquitin10 promoter"
     misc_difference 1481
                     /replace="g"
                     /label=killed PmlI (CACcTG)
                     /note="killed PmlI (CACcTG)"
     misc_feature    1662..1965
                     /label=cryptic intron from Norris et al. 1993
                     /note="cryptic intron from Norris et al. 1993"
     misc_difference 1894
                     /replace="t"
                     /label=killed XbaI (aCTAGA)
                     /note="killed XbaI (aCTAGA)"
     primer_bind     1966..1990
                     /label=pDOE mcs1F with XhoI
                     /note="pDOE mcs1F with XhoI"
     misc_feature    1978..1994
                     /label=XhoSac linker
                     /note="XhoSac linker"
     misc_feature    1995..2060
                     /label=TMV omega prime translation leader
                     /note="TMV omega prime translation leader"
     misc_feature    2005..2059
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     CDS             2061..2828
                     /codon_start=1
                     /gene="X-mTq2"
                     /product="mcs1-mTurquoise2 fusion protein"
                     /label=X-mTq2
                     /note="normalized for Arabidopsis"
                     /protein_id="AIN46673.1"
                     /translation="MGRALGTSGGSGGGSGMVSKGEELFTGVVPILVELDGDVNGHKFS
                     VSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSA
                     MPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYF
                     SDNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQS
                     KLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     gene            2061..2828
                     /gene="X-mTq2"
                     /label=X-mTq2
     misc_feature    2061..2108
                     /label=MCS1 front end
                     /note="MCS1 front end"
     misc_feature    2109..2828
                     /gene="X-mTq2"
                     /note="similar to protein sequence for mTurquoise2 crystal 
                     structure PDB 3ZTF; characterized in PubMed ID 22434194; 
                     mTq2, mTurquoise2 specific sequence"
     primer_bind     complement(2193..2215)
                     /label=mTq2 85R, binds mTq2 not mVenus
                     /note="mTq2 85R, binds mTq2 not mVenus"
     misc_difference 2547..2549
                     /gene="X-mTq2"
                     /label=differs from mTurquoise, mTq2=I146F
                     /note="differs from mTurquoise, mTq2=I146F"
     misc_feature    2696..2721
                     /gene="X-mTq2"
                     /label=mTq2 613F binds mTq2 but not mTurq or mVenus
                     /note="mTq2 613F binds mTq2 but not mTurq or mVenus"
     misc_feature    2829..2850
                     /label=MCS1 back end
                     /note="MCS1 back end"
     regulatory      2851..3575
                     /regulatory_class="terminator"
                     /note="OCS3octapine synthase terminator"
     terminator      2861..3568
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    2861..2869
                     /note="similar to native ocs region; c is 14nt downstream 
                     of Agro OCS stop codon (MacDonald 1991)"
     regulatory      3589..4226
                     /regulatory_class="promoter"
                     /note="UBQ10; ubiquitin10 promoter"
     misc_difference 3742
                     /replace="g"
                     /label=killed PmlI (CACcTG)
                     /note="killed PmlI (CACcTG)"
     misc_difference 4155
                     /replace="t"
                     /label=killed XbaI (aCTAGA)
                     /note="killed XbaI (aCTAGA)"
     primer_bind     4227..4251
                     /label=pDOE mcs3bF with Bsu36I
                     /note="pDOE mcs3bF with Bsu36I"
     misc_feature    4234..4249
                     /label=Bsu361 linker
                     /note="Bsu361 linker"
     regulatory      4250..4315
                     /regulatory_class="enhancer"
                     /note="TMV-Omega enhancer"
     misc_feature    4260..4314
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     CDS             4316..5098
                     /codon_start=1
                     /gene="mVenus-X"
                     /product="mVenus-mcs3 fusion protein"
                     /label=mVenus-X
                     /protein_id="AIN46674.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSGAGSGGGSGMGSLRSHVTSG"
     gene            4316..5098
                     /gene="mVenus-X"
                     /label=mVenus-X
     misc_feature    4316..5032
                     /gene="mVenus-X"
                     /note="similar to protein sequence AAZ65844.1; 
                     characterized in PubMed ID 16716067; mVenus"
     CDS             4316..5032
                     /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 4934..4936
                     /gene="mVenus-X"
                     /label=A206K monomerizing mutation
                     /note="A206K monomerizing mutation"
     misc_feature    5033..5098
                     /label=MCS3 front end
                     /note="MCS3 front end"
     misc_feature    5099..5116
                     /label=MCS3 back end
                     /note="MCS3 back end"
     regulatory      5118..5370
                     /regulatory_class="terminator"
                     /note="NOS nopaline synthase terminator"
     terminator      5118..5370
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5380..5396
                     /label=remnant from pFGC5941
                     /note="remnant from pFGC5941"
     misc_feature    5397..11919
                     /label=derived from pCAMBIA1300 via pFGC5941
                     /note="derived from pCAMBIA1300 via pFGC5941"
     primer_bind     complement(5421..5437)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   5639..5662
                     /label=right border from C58 T-DNA
                     /note="right border from C58 T-DNA"
     misc_feature    5640..5664
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      6725..7725
                     /label=pVS1
                     /note="pVS1"
     CDS             6965..7594
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8023..9096
                     /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      9162..9356
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    9700..9840
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(10026..10614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      10129..10410
                     /label=pBR325
                     /note="pBR325"
     CDS             complement(10701..11495)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside phosphotransferase"
                     /function="kanamycin resistance"
                     /label=aadA
                     /note="AphA-3"
                     /protein_id="AIN46675.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(10701..11495)
                     /gene="aadA"
                     /label=aadA