LOCUS       40924_29401        6247 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-R3-TagRFP-T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6247)
  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 6247)
  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 6247)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6247)
  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..6247
                     /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(3016..3804)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     misc_feature    4002..4025
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(4079..5224)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     rep_origin      complement(5617..6148)
                     /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 tgttatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
     3061 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
     3121 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     3181 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     3241 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
     3301 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     3361 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     3421 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     3481 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     3541 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     3601 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     3661 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     3721 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     3781 ttcggcgatc accgcttccc tcatgatgtt taactttgtt ttagggcgac tgccctgctg
     3841 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
     3901 cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag ccatgaaaac
     3961 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt cgatcggatt ggcggttatg
     4021 cggttgcgat gcaggtggct gctgaacccc cagccggaac tgaccccaca aggccctagc
     4081 gtttgcaatg caccaggtca tcattgaccc aggcgtgttc caccaggccg ctgcctcgca
     4141 actcttcgca ggcttcgccg acctgctcgc gccacttctt cacgcgggtg gaatccgatc
     4201 cgcacatgag gcggaaggtt tccagcttga gcgggtacgg ctcccggtgc gagctgaaat
     4261 agtcgaacat ccgtcgggcc gtcggcgaca gcttgcggta cttctcccat atgaatttcg
     4321 tgtagtggtc gccagcaaac agcacgacga tttcctcgtc gatcaggacc tggcaacggg
     4381 acgttttctt gccacggtcc aggacgcgga agcggtgcag cagcgacacc gattccaggt
     4441 gcccaacgcg gtcggacgtg aagcccatcg ccgtcgcctg taggcgcgac aggcattcct
     4501 cggccttcgt gtaataccgg ccattgatcg accagcccag gtcctggcaa agctcgtaga
     4561 acgtgaaggt gatcggctcg ccgatagggg tgcgcttcgc gtactccaac acctgctgcc
     4621 acaccagttc gtcatcgtcg gcccgcagct cgacgccggt gtaggtgatc ttcacgtcct
     4681 tgttgacgtg gaaaatgacc ttgttttgca gcgcctcgcg cgggattttc ttgttgcgcg
     4741 tggtgaacag ggcagagcgg gccgtgtcgt ttggcatcgc tcgcatcgtg tccggccacg
     4801 gcgcaatatc gaacaaggaa agctgcattt ccttgatctg ctgcttcgtg tgtttcagca
     4861 acgcggcctg cttggcttcg ctgacctgtt ttgccaggtc ctcgccggcg gtttttcgct
     4921 tcttggtcgt catagttcct cgcgtgtcga tggtcatcga cttcgccaaa cctgccgcct
     4981 cctgttcgag acgacgcgaa cgctccacgg cggccgatgg cgcgggcagg gcagggggag
     5041 ccagttgcac gctgtcgcgc tcgatcttgg ccgtagcttg ctggactatc gagccgacgg
     5101 actggaaggt ttcgcggggc gcacgcatga cggtgcggct tgcgatggtt tcggcatcct
     5161 cggcggaaaa ccccgcgtcg atcagttctt gcctgtatgc cttccggtca aacgtccgat
     5221 tcattcaccc tccttgcggg attgccccgg aattaattcc ccggatcgat ccgtcgatct
     5281 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
     5341 cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa
     5401 aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc
     5461 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
     5521 tttctgcgga ctggctttct acgtggctgc catttttggg gtgaggccgt tcgcggccga
     5581 ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg ggccgggagg gttcgagaag
     5641 ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc tggttaaaaa
     5701 caaggtttat aaatattggt ttaaaagcag gttaaaagac aggttagcgg tggccgaaaa
     5761 acgggcggaa acccttgcaa atgctggatt ttctgcctgt ggacagcccc tcaaatgtca
     5821 ataggtgcgc ccctcatctg tcagcactct gcccctcaag tgtcaaggat cgcgcccctc
     5881 atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc cccaggcttg
     5941 tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt tgcgaggctg
     6001 gccagctcca cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac gccgcgccgg
     6061 gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg gccaagtttt
     6121 ccgcgaggta tccacaacgc cggcggccct acatggctct gctgtagtga gtgggttgcg
     6181 ctccggcagc ggtcctgatc ccccgcagaa aaaaaggatc tcaagaagat cctttgatct
     6241 tttctac
//
