LOCUS       Exported                3165 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-B4, complete sequence.
ACCESSION   KY825148
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3165)
  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 3165)
  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 3165)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3165
                     /organism="Cloning vector pLX-B4"
                     /mol_type="other DNA"
                     /db_xref="taxon:2016270"
     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"
     CDS             complement(299..475)
                     /codon_start=1
                     /product="beta-galactosidase alpha subunit"
                     /label=beta-galactosidase alpha subunit
                     /note="LacZ alpha peptide"
                     /protein_id="ASG92137.1"
                     /translation="MITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDR
                     PSQQLRSLNGEWR"
     regulatory      complement(522..579)
                     /regulatory_class="promoter"
                     /note="lacZ"
     promoter        complement(527..557)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_feature    653..676
                     /label=RB
                     /note="RB"
     regulatory      complement(741..781)
                     /regulatory_class="terminator"
     misc_feature    791..814
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(869..1402)
                     /codon_start=1
                     /gene="acc3"
                     /product="gentamicin-3-acetyltransferase"
                     /label=acc3
                     /note="AacC1"
                     /protein_id="ASG92138.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(869..1402)
                     /gene="acc3"
                     /label=acc3
     promoter        complement(1591..1619)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    1620..1643
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(1644..2306)
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /protein_id="ASG92139.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     gene            complement(1644..2306)
                     /gene="rep"
                     /label=rep
     rep_origin      2307..3078
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"