LOCUS       V004874                 7308 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004874
VERSION     V004874
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7308)
  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 7308)
  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 7308)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7308)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
            Biol (2017) In press"
            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"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      27..58
                     /regulatory_class="terminator"
     misc_feature    59..83
                     /label="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(106..129)
                     /note="left border of left T-DNA; LB"
     promoter        179..559
                     /label="MAS promoter"
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             570..1283
                     /label="TagRFP-T"
                     /note="photostable variant of TagRFP (Shaner et al., 2008)"
     mat_peptide     1281..1355
                     /product="T2A"
     CDS             1302..1355
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1356..3818
                     /gene="alcR"
                     /label="Regulatory protein alcR"
                     /note="Regulatory protein alcR from Emericella nidulans
                     (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 /
                     M139). Accession#: P21228"
     terminator      3824..4076
                     /label="MAS terminator"
                     /note="mannopine synthase terminator"
     misc_feature    4131..4154
                     /label="RB"
                     /note="RB"
     regulatory      complement(4225..4277)
                     /regulatory_class="terminator"
     CDS             complement(4341..4871)
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     promoter        complement(5060..5088)
                     /label="Pc promoter"
                     /note="class 1 integron promoter"
     misc_feature    5089..5112
                     /label="assembly linker"
                     /note="assembly linker"
     CDS             complement(5166..6311)
                     /label="trfA"
                     /note="trans-acting replication protein that binds to and
                     activates oriV"
     rep_origin      complement(6704..7235)
                     /direction=LEFT
                     /label="oriV"
                     /note="incP origin of replication"
ORIGIN
        1 caagaagatc ctttgatctt ttctacaggc ctgctggtaa tcgcaggcct ttttattttg
       61 gcaggatata ttgtggtgta aacacttacc gcacctctgc agcagcggca ggatatatgg
      121 cagtgtaaac tccattttcg aacgcgttaa ttaagtagcc tggtcgaaac cgtctcattt
      181 ttcaaatcag tgcgcaagac gtgacgtaag tatccgagtc agtttttatt tttctactaa
      241 tttggtcgtt tatttcggcg tgtaggacat ggcaaccggg cctgaatttc gcgggtattc
      301 tgtttctatt ccaacttttt cttgatccgc agccattaac gacttttgaa tagatacgct
      361 gacacgccaa gcctcgctag tcaaaagtgt accaaacaac gctttacagc aagaacggaa
      421 tgcgcgtgac gctcgcggtg acgccatttc gccttttcag aaatggataa atagccttgc
      481 ttcctattat atcttcccaa attaccaata cattacacta gcatctgaat ttcataacca
      541 atctcgatac accaaatcgg gtaacaacaa tggtttcaaa gggagaagag ctgattaagg
      601 agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca
      661 catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg
      721 agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca
      781 gaaccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccctgagg
      841 gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg
      901 acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc
      961 catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccaac accgagatgc
     1021 tgtaccccgc tgacggcggc ctggaaggca gaacagacat ggccctgaag ctcgtgggcg
     1081 ggggccacct gatctgcaac ttcaagacca catacagatc caagaaaccc gctaagaacc
     1141 tcaagatgcc cggcgtctac tatgtggacc acagactgga aagaatcaag gaggccgaca
     1201 aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca
     1261 aactggggca caaacttaac ggaggcggag gcgggagagc agaagggaga ggaagcttgc
     1321 taacctgtgg agacgttgag gaaaatccag ggccaatggc agatacacgc cgacgccaga
     1381 atcatagctg cgatccctgt cgcaagggca agcgacgctg tgatgccccg gaaaatagaa
     1441 acgaggccaa tgaaaacggc tgggtttcgt gttcaaattg caagcgttgg aacaaggatt
     1501 gtaccttcaa ttggctctca tcccaacgct ccaaggcaaa aggggctgca cctagagcga
     1561 gaacaaagaa agccaggacc gcaacaacca ccagtgaacc atcaacttca gctgcaacaa
     1621 tccctacacc ggaaagtgac aatcacgatg cgcctccagt cataaactct cacgacgcgc
     1681 tcccgagctg gactcagggg ctactctccc accccggcga ccttttcgat ttcagccact
     1741 ctgctattcc cgcaaatgca gaagatgcgg ccaacgtgca gtcagacgca ccttttccgt
     1801 gggatctagc catccccggt gatttcagca tgggccaaca gctcgagaaa cctctcagtc
     1861 cgctcagttt tcaagcagtc cttcttccgc cccatagccc gaacacggat gacctcattc
     1921 gcgagctgga agagcagact acagatccgg actcggttac cgatactaat agtgtacaac
     1981 aggtcgctca agatggatcg ctatggtctg atcggcagtc gccgctactg cctgagaaca
     2041 gtctgtgcat ggcctcagac agcacagcac ggcgatatgc ccgttccaca atgacgaaga
     2101 atctgatgcg aatctaccac gatagtatgg agaatgcact gtcctgctgg ctgacagagc
     2161 acaattgtcc atactccgac cagatcagct acctgccgcc caagcagcgg gcggaatggg
     2221 gcccgaactg gtcaaacagg atgtgcatcc gggtgtgccg gctagatcgc gtatctacct
     2281 cattacgcgg gcgcgccctg agtgcggaag aggacaaagc cgcagcccga gccctgcatc
     2341 tggcgatcgt agcttttgcg tcgcaatgga cgcagcatgc gcagaggggg gctgggctaa
     2401 atgttcctgc agacatagcc gccgatgaga ggtccatccg gaggaacgcc tggaatgaag
     2461 cacgccatgc cttgcagcac acgacaggga ttccatcatt ccgggttata tttgcgaata
     2521 tcatcttttc tctcacgcaa tctgtgctgg atgatgatga gcagcacggt atgggtgcac
     2581 gtctagacaa gctactcgaa aatgacggtg cgcccgtgtt cctggaaacc gcgaaccgtc
     2641 agctttatac attccgacat aagtttgcac gaatgcaacg ccgcggtaag gctttcaaca
     2701 ggctcccggg aggatctgtc gcatcgacat tcgccggtat tttcgagaca ccgacgccgt
     2761 cgtctgaaag cccacagctt gacccggttg tggccagtga ggagcatcgc agtacattaa
     2821 gccttatgtt ctggctaggg atcatgttcg atacactaag cgctgcaatg taccagcgac
     2881 cactcgtggt gtcagatgag gatagccaga tatcatcggc atctccacca aggcgcggcg
     2941 ctgaaacgcc gatcaaccta gactgctggg agcccccgag acaggtcccg agcaatcaag
     3001 aaaagagcga cgtatggggc gacctcttcc tccgcacctc ggactctctc ccagatcacg
     3061 aatcccacac acaaatctct cagccagcgg ctcgatggcc ctgcacctac gaacaggccg
     3121 ccgccgctct ctcctctgca acgcccgtca aagtcctcct ctaccgccgc gtcacgcagc
     3181 tccaaaccct cctctatcgc ggcgccagcc ctgcccgcct tgaagcggcc atccagagaa
     3241 cgctctacgt ttataatcac tggacagcga agtaccaacc atttatgcag gactgcgttg
     3301 ctaaccacga gctcctccct tcgcgcatcc agtcttggta cgtcattcta gacggtcact
     3361 ggcatctagc cgcgatgttg ctagcggacg ttttggagag catcgaccgc gattcgtact
     3421 ctgatatcaa ccacatcgac cttgtaacaa agctaaggct cgataatgca ctagcagtta
     3481 gtgcccttgc gcgctcttca ctccgaggcc aggagctgga cccgggcaaa gcatctccga
     3541 tgtatcgcca tttccatgat tctctgaccg aggtggcatt cctggtagaa ccgtggaccg
     3601 tcgttcttat tcactcgttt gccaaagctg cgtatatctt gctggactgt ttagatctgg
     3661 acggccaagg aaatgcacta gcggggtacc tgcagctgcg gcaaaattgc aactactgca
     3721 ttcgggcgct gcaatttctg ggcaggaagt cggatatggc ggcgctggtt gcgaaggatt
     3781 tagagagagg tttgaatggg aaagttgaca gcttcttgta gctcttggac tcccatgttg
     3841 gcaaaggcaa ccaaacaaac aatgaatgat ccgctcctgc atatggggcg gtttgagtat
     3901 ttcaactgcc atttgggctg aattgaagac atgctcctgt cagaaattcc gtgatcttac
     3961 tcaatattca gtaatctcgg ccaatatcct aaatgtgcgt ggctttatct gtctttgtat
     4021 tgtttcatca attcatgtaa cgtttgcttt tcttatgaat tttcaaataa attatcgtca
     4081 ggagacggga caaggatgcg cctgcaggtt gttgatgatg tgatgactga tggcaggata
     4141 tatgtggttg taattcattt ctaccgtgta atttactgta tttttttgtt tgttcgttcg
     4201 tttgtaaaaa tattttttgg aagcaaaaaa ttagcgcaag aagacaaaaa tcaccttgcg
     4261 ctaatgctct gttacaggcg cgccaattta cccaacaact ccgcggccgg gaagccgatc
     4321 tcggcttgaa cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct
     4381 tcccgtatgc ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc
     4441 acgtagatca cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg
     4501 gtggcaatgc cctgcctccg gtgctctccg gagactgcga gatcatagat atagatctca
     4561 ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct
     4621 tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg
     4681 gagtccggct gatgttggga gtaggtggct acatcgccga actcacgacc gaaaagatca
     4741 agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc
     4801 atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg
     4861 ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg
     4921 ctgcttggat gcccgaggca taggctgtac aaaaaaacag tcataacaag ccatgaaaac
     4981 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     5041 catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaaga tcggattggc
     5101 ggttatgcgg ttgcgatgca ggtggctgct gaacccccag ccggaactga ccccacaagg
     5161 ccctagcgtt tgcaatgcac caggtcatca ttgacccagg cgtgttccac caggccgctg
     5221 cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa
     5281 tccgatccgc acatgaggcg gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag
     5341 ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct tgcggtactt ctcccatatg
     5401 aatttcgtgt agtggtcgcc agcaaacagc acgacgattt cctcgtcgat caggacctgg
     5461 caacgggacg ttttcttgcc acggtccagg acgcggaagc ggtgcagcag cgacaccgat
     5521 tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg
     5581 cattcctcgg ccttcgtgta ataccggcca ttgatcgacc agcccaggtc ctggcaaagc
     5641 tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta ctccaacacc
     5701 tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc
     5761 acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg
     5821 ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc
     5881 ggccacggcg caatatcgaa caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt
     5941 ttcagcaacg cggcctgctt ggcttcgctg acctgttttg ccaggtcctc gccggcggtt
     6001 tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct
     6061 gccgcctcct gttctagccg acgcgaacgc tccacggcgg ccgatggcgc gggcagggca
     6121 gggggagcca gttgcacgct gtcgcgctcg atcttggccg tagcttgctg gactatcgag
     6181 ccgacggact ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg
     6241 gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac
     6301 gtccgattca ttcaccctcc ttgcgggatt gccccggaat taattccccg gatcgatccg
     6361 tcgatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
     6421 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg
     6481 tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc
     6541 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
     6601 agcaccgttt ctgcggactg gctttctacg tggctgccat ttttggggtg aggccgttcg
     6661 cggccgaggg gcgcagcccc tggggggatg ggaggcccgc gttagcgggc cgggagggtt
     6721 cgagaagggg gggcaccccc cttcggcgtg cgcggtcacg cgcacagggc gcagccctgg
     6781 ttaaaaacaa ggtttataaa tattggttta aaagcaggtt aaaagacagg ttagcggtgg
     6841 ccgaaaaacg ggcggaaacc cttgcaaatg ctggattttc tgcctgtgga cagcccctca
     6901 aatgtcaata ggtgcgcccc tcatctgtca gcactctgcc cctcaagtgt caaggatcgc
     6961 gcccctcatc tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc acttatcccc
     7021 aggcttgtcc acatcatctg tgggaaactc gcgtaaaatc aggcgttttc gccgatttgc
     7081 gaggctggcc agctccacgt cgccggccga aatcgagcct gcccctcatc tgtcaacgcc
     7141 gcgccgggtg agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc agtgagggcc
     7201 aagttttccg cgaggtatcc acaacgccgg cggccctaca tggctctgct gtagtgagtg
     7261 ggttgcgctc cggcagcggt cctgatcccc cgcagaaaaa aaggatct
//
