LOCUS       40924_29391        6185 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-R2-TagRFP-T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6185)
  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 6185)
  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 6185)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6185)
  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..6185
                     /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"
     promoter        535..1204
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     5'UTR           1204..1349
                     /label=PPV
                     /note="PPV"
     CDS             1350..2060
                     /label=TagRFP-T
                     /note="photostable variant of TagRFP (Shaner et al., 2008)"
     3'UTR           2236..2452
                     /label=PPV
                     /note="PPV"
     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(3026..3838)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    3940..3963
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(4017..5162)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     rep_origin      complement(5555..6086)
                     /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 accgtctcac cagtacgcac gattcaaggc ttgcttcaca aaccaaggca
      301 agtaatagag attggagtct ctaaaaaggt agttcccact gaatcaaagg ccatggagtc
      361 aaagattcaa atagaggacc taacagaact cgccgtaaag actggcgaac agttcataca
      421 gagtctctta cgactcaatg acaagaagaa aatcttcgtc aacatggtgg agcacgacac
      481 acttgtctac tccaaaaata tcaaagatac agtctcagaa gaccaaaggg caattgagac
      541 ttttcaacaa agggtaatat ccggaaacct cctcggattc cattgcccag ctatctgtca
      601 ctttattgtg aagatagtgg aaaaggaagg tggctcctac aaatgccatc attgcgataa
      661 aggaaaggcc atcgttgaag atgcctctgc cgacagtggt cccaaagatg gacccccacc
      721 cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg
      781 atgtgataac atggtggagc acgacacact tgtctactcc aaaaatatca aagatacagt
      841 ctcagaagac caaagggcaa ttgagacttt tcaacaaagg gtaatatccg gaaacctcct
      901 cggattccat tgcccagcta tctgtcactt tattgtgaag atagtggaaa aggaaggtgg
      961 ctcctacaaa tgccatcatt gcgataaagg aaaggccatc gttgaagatc ctctgccgac
     1021 agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca
     1081 accacgtctt caaagcaagt ggattgatgt gatatctcca ctgacgtaag ggatgacgca
     1141 caatcccact atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag
     1201 aggaaaatat aaaaactcaa cacaacatac aaaattttaa gcaatcaaat caatctcaag
     1261 ctatcaaaat ttttcaaatc tcacttgaaa gatcaaaaat caacaaagaa aatcccttaa
     1321 tttctctacc aaatttactg caagtcaaga tggtttcaaa gggagaagag ctgattaagg
     1381 agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca
     1441 catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg
     1501 agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca
     1561 gaaccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccctgagg
     1621 gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg
     1681 acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc
     1741 catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccaac accgagatgc
     1801 tgtaccccgc tgacggcggc ctggaaggca gaacagacat ggccctgaag ctcgtgggcg
     1861 ggggccacct gatctgcaac ttcaagacca catacagatc caagaaaccc gctaagaacc
     1921 tcaagatgcc cggcgtctac tatgtggacc acagactgga aagaatcaag gaggccgaca
     1981 aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca
     2041 aactggggca caaacttaac taatcgatca cgtgaagctc atatccagat gaaggcagca
     2101 gcattgagaa atgttcaaaa tcgtttattt ggcttggatg gaaacgtcgg aacacaagaa
     2161 gaggacacag agagacacac cgctggtgat gttaatcgca acatgcacaa cctcctcggt
     2221 gtgaggggag tgtagtggtc tcggtatcta tcataaactc tacctgggtg agagtctaat
     2281 catccagttg tttttagatt cctgttagca tccttttctc cgctttaata gcagtacatt
     2341 cagtgaggtt ttacctccat atgttctagt ctgttattgt cgaacacagg cccttgtatc
     2401 tgatgtagcg agtgcttcac tccattcggg ttatagttct tgtgcaagag acaaaaaaaa
     2461 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2521 atgcatgcct gcagatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt
     2581 tgccggtctt gcgatgatta tcatctaatt tctgttgaat tacgttaagc atgtaataat
     2641 taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt
     2701 atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg
     2761 cgcggtgtca tctatgttac tcgatctcgt caggagacgg gacaaggatg cgagctagcc
     2821 tgcaggaatt gttgattttg tgatgactga tggcaggata tatgcggttg taattcattt
     2881 ttattgtcta aatttctgta tttgtttgtt tgttcggttg taaatttttt tggaagaaca
     2941 agaaaagaaa aaacacccgt tagggtgttt ttagttagtg tggcgcgccg acttgcgaca
     3001 tgcggtcctt tgcaatcaac tattagaaaa attcatccag catcagatga aattgcagtt
     3061 tgttcatatc cggattatca atgccatatt tctgaaacag acgtttttgc aggctcgggc
     3121 taaattcgcc caggcagttc cacagaatgg ccagatcctg ataacgatcc gcaatgccca
     3181 cacggcccac atcaatgcag ccaatcagtt tgccttcatc gaaaatcagg ttatccaggc
     3241 taaaatcgcc gtgggtcacc acgctatccg ggctaaacgg cagcagttta tgcatttctt
     3301 tccacacctg ttccaccggc cagccgttac gttcatcatc aaaatcgctc gcatccacca
     3361 ggccgttgtt catacggctc tgcgcctggg ccagacgaaa cacacgatcg ctgttaaacg
     3421 ggcagttgca caccggaatg ctatgcagac gacgcagaaa cacggccagc gcatccacaa
     3481 tgttttcgcc gctatccgga tattcttcca gcacctgaaa cgcggttttg cccggaatcg
     3541 cggtggtcag cagccacgca tcatccgggg tgcgaataaa atgtttaatg gtcggcagcg
     3601 gcataaattc ggtcagccag ttcagacgca ccatttcatc ggtcacatcg ttcgccacgc
     3661 tgcctttgcc atgtttcaga aacagttccg gcgcatccgg tttgccatac agacgataaa
     3721 tggtcgcgcc gctctgaccc acgttatcac gcgcccattt atagccatac agatccgcat
     3781 ccatgttgct gttcagacgc ggacggctac agctcgtttc acgctgaata tggctcataa
     3841 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     3901 tatcttgtgc aatgtaacat cagagatttt gagacacaag atcggattgg cggttatgcg
     3961 gttgcgatgc aggtggctgc tgaaccccca gccggaactg accccacaag gccctagcgt
     4021 ttgcaatgca ccaggtcatc attgacccag gcgtgttcca ccaggccgct gcctcgcaac
     4081 tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga atccgatccg
     4141 cacatgaggc ggaaggtttc cagcttgagc gggtacggct cccggtgcga gctgaaatag
     4201 tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact tctcccatat gaatttcgtg
     4261 tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga tcaggacctg gcaacgggac
     4321 gttttcttgc cacggtccag gacgcggaag cggtgcagca gcgacaccga ttccaggtgc
     4381 ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag gcattcctcg
     4441 gccttcgtgt aataccggcc attgatcgac cagcccaggt cctggcaaag ctcgtagaac
     4501 gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt actccaacac ctgctgccac
     4561 accagttcgt catcgtcggc ccgcagctcg acgccggtgt aggtgatctt cacgtccttg
     4621 ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg ggattttctt gttgcgcgtg
     4681 gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc cggccacggc
     4741 gcaatatcga acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg tttcagcaac
     4801 gcggcctgct tggcttcgct gacctgtttt gccaggtcct cgccggcggt ttttcgcttc
     4861 ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc tgccgcctcc
     4921 tgttcgagac gacgcgaacg ctccacggcg gccgatggcg cgggcagggc agggggagcc
     4981 agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct ggactatcga gccgacggac
     5041 tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc ggcatcctcg
     5101 gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa cgtccgattc
     5161 attcaccctc cttgcgggat tgccccggaa ttaattcccc ggatcgatcc gtcgatcttg
     5221 atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct
     5281 tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct gtccataaaa
     5341 ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt ctctttgcgc
     5401 ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt
     5461 tctgcggact ggctttctac gtggctgcca tttttggggt gaggccgttc gcggccgagg
     5521 ggcgcagccc ctggggggat gggaggcccg cgttagcggg ccgggagggt tcgagaaggg
     5581 ggggcacccc ccttcggcgt gcgcggtcac gcgcacaggg cgcagccctg gttaaaaaca
     5641 aggtttataa atattggttt aaaagcaggt taaaagacag gttagcggtg gccgaaaaac
     5701 gggcggaaac ccttgcaaat gctggatttt ctgcctgtgg acagcccctc aaatgtcaat
     5761 aggtgcgccc ctcatctgtc agcactctgc ccctcaagtg tcaaggatcg cgcccctcat
     5821 ctgtcagtag tcgcgcccct caagtgtcaa taccgcaggg cacttatccc caggcttgtc
     5881 cacatcatct gtgggaaact cgcgtaaaat caggcgtttt cgccgatttg cgaggctggc
     5941 cagctccacg tcgccggccg aaatcgagcc tgcccctcat ctgtcaacgc cgcgccgggt
     6001 gagtcggccc ctcaagtgtc aacgtccgcc cctcatctgt cagtgagggc caagttttcc
     6061 gcgaggtatc cacaacgccg gcggccctac atggctctgc tgtagtgagt gggttgcgct
     6121 ccggcagcgg tcctgatccc ccgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6181 tctac
//
