LOCUS       Exported                5547 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-B3-TagRFP-T, complete sequence.
ACCESSION   KY825147
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5547)
  AUTHORS   Pasin F, Bedoya LC, Bernabe-Orts JM, Gallo A, Simon-Mateo C, Orzaez 
            D, Garcia JA.
  TITLE     Multiple T-DNA Delivery to Plants Using Novel Mini Binary Vectors 
            with Compatible Replication Origins
  JOURNAL   ACS Synth Biol (2017) In press
  PUBMED    28657330
REFERENCE   2  (bases 1 to 5547)
  AUTHORS   Pasin F, Garcia JA.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAR-2017) Department of Plant Molecular Genetics, 
            Centro Nacional de Biotecnologia (CNB-CSIC), Darwin 3, Madrid, 
            Madrid 28049, Spain
REFERENCE   3  (bases 1 to 5547)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5547
                     /organism="Cloning vector pLX-B3-TagRFP-T"
                     /mol_type="other DNA"
                     /db_xref="taxon:2016267"
     misc_feature    1..24
                     /label=assembly linker
                     /note="assembly linker"
     regulatory      25..72
                     /regulatory_class="terminator"
     misc_feature    133..157
                     /label=LB
                     /note="LB"
     misc_feature    180..210
                     /label=LB
                     /note="LB"
     misc_feature    184..208
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      260..1203
                     /regulatory_class="promoter"
                     /note="2x CaMV 35S"
     promoter        862..1206
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     5UTR           1204..1349
                     /label=PPV
                     /note="PPV"
     CDS             1350..2063
                     /codon_start=1
                     /product="TagRFP"
                     /label=TagRFP
                     /note="derived from eqFP578, from the sea anemone Entacmaea
                     quadricolor], a bright monomeric red fluorescent protein 
                     with excitation/emission peaks at 555/584 nm, 2.8-fold 
                     brighter fluorescence than mCherry, the best monomeric red 
                     fluorescent protein reported to date (Merzlyak et al., 
                     2007)"
                     /protein_id="ASG92134.1"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     3UTR           2236..2452
                     /label=PPV
                     /note="PPV"
     regulatory      2531..2786
                     /regulatory_class="terminator"
                     /note="nos"
     terminator      2534..2786
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2851..2874
                     /label=RB
                     /note="RB"
     regulatory      complement(2939..2979)
                     /regulatory_class="terminator"
     misc_feature    2989..3012
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(3013..3918)
                     /codon_start=1
                     /gene="aadA"
                     /product="spectinomycin adenyltransferase"
                     /label=aadA
                     /note="AadA"
                     /protein_id="ASG92135.1"
                     /translation="MPRASKQQARYAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIA
                     EVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRAL
                     INDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPA
                     TIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDER
                     NVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRA
                     DQLEEFVHYVKGEITKVVGK"
     gene            complement(3013..3918)
                     /gene="aadA"
                     /label=aadA
     CDS             complement(3013..3804)
                     /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"
     misc_feature    4002..4025
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(4026..4688)
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /protein_id="ASG92136.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     gene            complement(4026..4688)
                     /gene="rep"
                     /label=rep
     rep_origin      4689..5460
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"