LOCUS       40924_29411        6063 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-R4-TagRFP-T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6063)
  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 6063)
  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 6063)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6063)
  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..6063
                     /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(3070..3600)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(3789..3817)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    3818..3841
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(3895..5040)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     rep_origin      complement(5433..5964)
                     /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 tgcaatttac ccaacaactc cgcggccggg aagccgatct cggcttgaac
     3061 gaattgttag gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc
     3121 caactttgta tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac
     3181 ataagcacca agcgcgttgg cctcatgctt gaggagattg atgagcgcgg tggcaatgcc
     3241 ctgcctccgg tgctctccgg agactgcgag atcatagata tagatctcac tacgcggctg
     3301 ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc
     3361 agcaagcgcg atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg
     3421 atgttgggag taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg
     3481 catggatttg acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc
     3541 acctaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat
     3601 cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg
     3661 cccgaggcat aggctgtaca aaaaaacagt cataacaagc catgaaaacc gccactgcgc
     3721 cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact
     3781 tgcattacag tttacgaacc gaacaggctt atgtcaagat cggattggcg gttatgcggt
     3841 tgcgatgcag gtggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt
     3901 gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc
     3961 ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca
     4021 catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc
     4081 gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta
     4141 gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt
     4201 tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc
     4261 aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc
     4321 cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt
     4381 gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac
     4441 cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt
     4501 gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt
     4561 gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc
     4621 aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc
     4681 ggcctgcttg gcttcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt
     4741 ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg
     4801 ttcgagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag
     4861 ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg actatcgagc cgacggactg
     4921 gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc
     4981 ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat
     5041 tcaccctcct tgcgggattg ccccggaatt aattccccgg atcgatccgt cgatcttgat
     5101 cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc
     5161 ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt ccataaaacc
     5221 gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct ctttgcgctt
     5281 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     5341 tgcggactgg ctttctacgt ggctgccatt tttggggtga ggccgttcgc ggccgagggg
     5401 cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg
     5461 ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag
     5521 gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg
     5581 gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag
     5641 gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct
     5701 gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca
     5761 catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca
     5821 gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga
     5881 gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc
     5941 gaggtatcca caacgccggc ggccctacat ggctctgctg tagtgagtgg gttgcgctcc
     6001 ggcagcggtc ctgatccccc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6061 tac
//
