LOCUS       Exported               10171 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pKT pKT-NB-H2BsfGFP, complete sequence.
ACCESSION   MH973511
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10171)
  AUTHORS   Kakimoto T, Tsujimura K.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-SEP-2018) Department of Biological Sciences, Graduate 
            School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, 
            Osaka 560-0043, Japan
REFERENCE   2  (bases 1 to 10171)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10171
                     /organism="Binary vector pKT pKT-NB-H2BsfGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:2485971"
     misc_feature    1..7579
                     /label=derived from pLHAB
                     /note="derived from pLHAB"
     misc_feature    279..303
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    279..302
                     /label=Left border of T-DNA
                     /note="Left border of T-DNA"
     CDS             complement(823..1833)
                     /codon_start=1
                     /product="streptomycin/spectinomycin adenyltransferase"
                     /label=streptomycin/spectinomycin adenyltransferase
                     /protein_id="AYR16914.1"
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(823..1614)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2219..2413
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2479..3552)
                     /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"
     CDS             complement(3981..4610)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    5945..6085
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6270..6858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    7503..7527
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    7503..7525
                     /label=Right Border of T-DNA
                     /note="Right Border of T-DNA"
     regulatory      7580..7872
                     /regulatory_class="promoter"
                     /note="NOS promoter"
     promoter        7663..7842
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             7893..8444
                     /codon_start=1
                     /product="phosphinothricin acetyltransferase"
                     /label=phosphinothricin acetyltransferase
                     /protein_id="AYR16911.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     regulatory      8462..8684
                     /regulatory_class="terminator"
                     /note="g7 terminator"
     terminator      8475..8675
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     primer_bind     complement(8688..8704)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8724..8761
                     /label=multi cloning site
                     /note="multi cloning site"
     CDS             8762..9925
                     /codon_start=1
                     /product="histone H2B-MCS-sfGFP fusion"
                     /label=histone H2B-MCS-sfGFP fusion
                     /protein_id="AYR16912.1"
                     /translation="MAKADKKPAEKKPAEKTPAAEPAAAAEKKPKAGKKLPKEPAGAGD
                     KKKKRSKKNVETYKIYIFKVLKQVHPDIGISSKAMGIMNSFINDIFEKLAGESSKLARY
                     NKKPTITSREIQTAVRLVLPGELAKHAVSEGTKAVTKFTSSGGGGMSKGEELFTGVVPI
                     LVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSR
                     YPDHMKRHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKE
                     DGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDG
                     PVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITHGMDELYK"
     misc_feature    9197..9208
                     /label=4x glycine linker
                     /note="4x glycine linker"
     CDS             9209..9925
                     /codon_start=1
                     /product="sfGFP"
                     /label=sfGFP
                     /note="codon optimized for Arabidopsis and removed possible
                     splicing sites"
                     /protein_id="AYR16913.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    9926..9940
                     /label=multi cloning site
                     /note="multi cloning site"
     regulatory      9941..10134
                     /regulatory_class="terminator"
                     /note="terminator of CaMV 35S"