LOCUS       40924_29416        3865 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-R4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3865)
  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 3865)
  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 3865)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3865)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-MAR-2017) Department of Plant Molecular Genetics, Centro 
            Nacional de Biotecnologia (CNB-CSIC), Darwin 3, Madrid, Madrid 
            28049, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..24
                     /label=assembly linker
                     /note="assembly linker"
     regulatory      25..72
                     /regulatory_class="terminator"
     misc_feature    133..157
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    184..208
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(308..478)
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     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(872..1402)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(1591..1619)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    1620..1643
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(1697..2842)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     rep_origin      complement(3235..3766)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 ccatcatcag ttcggtggtc ttccgacgaa caataaggcc gcaaatcgcg gcctttttta
       61 ttgataacaa aaccggctca gttctgcgta gaaaccaaca tgcaagctcc accgggtgca
      121 aagcggcagc ggcggcagga tatattcaat tgtaaatggc ttcatgtccg ggaaatctac
      181 tggtggcagg atatattgtg gtgtaaacaa tggagaaaaa gattaattaa gtactagtag
      241 cctggtcgaa accgtctcag gagagagacc aaaagcaaaa acccgccgaa gcgggttact
      301 agcgccattc gccattcaga gagcggagct gctgcgacgg acgatcggta cgcgcctctt
      361 cgctattacg ccaactggcg aaaggtggat gtgctgcaag gcgattaagt tgggtaacgc
      421 cagggttttc ccagtcacga cgttgtagta ccacggcaag gctatctgta atcattgttg
      481 tactccggtt aggacggatt gggaactggc taactcaaaa tccacacatt atacgagccg
      541 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag gtctctcgct gtcaggagac
      601 gggacaagga tgcgagctag cctgcaggaa ttgttgattt tgtgatgact gatggcagga
      661 tatatgcggt tgtaattcat ttttattgtc taaatttctg tatttgtttg tttgttcggt
      721 tgtaaatttt tttggaagaa caagaaaaga aaaaacaccc gttagggtgt ttttagttag
      781 tgtggcgcgc cgacttgcga catgcggtcc tttgcaattt acccaacaac tccgcggccg
      841 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
      901 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
      961 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     1021 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctctcc ggagactgcg agatcataga
     1081 tatagatctc actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa
     1141 caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc
     1201 cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac
     1261 cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc
     1321 gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa
     1381 catcgttgct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     1441 taacgcgctt gctgcttgga tgcccgaggc ataggctgta caaaaaaaca gtcataacaa
     1501 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     1561 ttgcgtgagc gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaag
     1621 atcggattgg cggttatgcg gttgcgatgc aggtggctgc tgaaccccca gccggaactg
     1681 accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca
     1741 ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca
     1801 cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct
     1861 cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact
     1921 tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga
     1981 tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca
     2041 gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta
     2101 ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt
     2161 cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt
     2221 actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt
     2281 aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg
     2341 ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc
     2401 gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct
     2461 gcttcgtgtg tttcagcaac gcggcctgct tggcttcgct gacctgtttt gccaggtcct
     2521 cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact
     2581 tcgccaaacc tgccgcctcc tgttcgagac gacgcgaacg ctccacggcg gccgatggcg
     2641 cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct
     2701 ggactatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg
     2761 cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct
     2821 tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccggaa ttaattcccc
     2881 ggatcgatcc gtcgatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat
     2941 ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg
     3001 ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc
     3061 tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     3121 catccggggt cagcaccgtt tctgcggact ggctttctac gtggctgcca tttttggggt
     3181 gaggccgttc gcggccgagg ggcgcagccc ctggggggat gggaggcccg cgttagcggg
     3241 ccgggagggt tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac gcgcacaggg
     3301 cgcagccctg gttaaaaaca aggtttataa atattggttt aaaagcaggt taaaagacag
     3361 gttagcggtg gccgaaaaac gggcggaaac ccttgcaaat gctggatttt ctgcctgtgg
     3421 acagcccctc aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc ccctcaagtg
     3481 tcaaggatcg cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa taccgcaggg
     3541 cacttatccc caggcttgtc cacatcatct gtgggaaact cgcgtaaaat caggcgtttt
     3601 cgccgatttg cgaggctggc cagctccacg tcgccggccg aaatcgagcc tgcccctcat
     3661 ctgtcaacgc cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc cctcatctgt
     3721 cagtgagggc caagttttcc gcgaggtatc cacaacgccg gcggccctac atggctctgc
     3781 tgtagtgagt gggttgcgct ccggcagcgg tcctgatccc ccgcagaaaa aaaggatctc
     3841 aagaagatcc tttgatcttt tctac
//
