LOCUS       Exported                3987 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-R2, complete sequence.
ACCESSION   KY825151
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3987)
  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 3987)
  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 3987)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3987
                     /organism="Cloning vector pLX-R2"
                     /mol_type="other DNA"
                     /db_xref="taxon:2016273"
     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="ASG92146.1"
                     /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(825..1640)
                     /codon_start=1
                     /gene="neo"
                     /product="neomycin phosphotransferase"
                     /label=neo
                     /note="kanamycin resistance protein; NptI"
                     /protein_id="ASG92147.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(825..1640)
                     /gene="neo"
                     /label=neo
     misc_feature    1742..1765
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(1816..2964)
                     /codon_start=1
                     /gene="trfA"
                     /product="replication protein TrfA"
                     /label=trfA
                     /protein_id="ASG92148.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(1816..2964)
                     /gene="trfA"
                     /label=trfA
     rep_origin      3357..3888
                     /label=oriV
                     /note="incP origin of replication"