LOCUS       40924_29316        6637 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-B2-NptII-DsRED, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6637)
  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 6637)
  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 6637)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6637)
  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..6637
                     /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"
     terminator      complement(287..539)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(582..1373)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(1407..1586)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2450..2794
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             2867..3541
                     /label=DsRed2
                     /note="improved tetrameric variant of DsRed fluorescent
                     protein"
     regulatory      complement(3549..3774)
                     /label=CaMV 35S
                     /note="CaMV 35S"
                     /regulatory_class="terminator"
     misc_feature    4003..4026
                     /label=RB
                     /note="RB"
     regulatory      complement(4091..4131)
                     /regulatory_class="terminator"
     misc_feature    4141..4164
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(4178..4990)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    5092..5115
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(5119..5778)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(5779..6550)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for 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 gagagcgatc agcttgcatg ccggtcgatc tagtaacata
      301 gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt ttctatcgcg
      361 tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa taacgtcatg
      421 cattacatgt taattattac atgcttaacg taattcaaca gaaattatat gataatcatg
      481 gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct
      541 gcttgactct agctagagtc cgaaccccag agtcccgctc agaagaactc gtcaagaagg
      601 cgatagaagg ctatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
      661 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
      721 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
      781 accatgatat tcggcaagca ggcgtcgccg tgggtcacga cgagatcctc gccgtcgggc
      841 atccgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
      901 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcggtgt
      961 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     1021 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     1081 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     1141 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttggagttca
     1201 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     1261 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     1321 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcctcga
     1381 tcgagttgag agtgaatatg agactctaat tggataccga ggggaattta tggaacgtca
     1441 gtggagcatt tttgacaaga aatatttgct agctgatagt gaccttaggc gacttttgaa
     1501 cgcgcaataa tggtttctga cgtatgtgct tagctcatta aactccagaa acccgcggct
     1561 gagtggctcc ttcaacgttg cggttctgtc agttccaaac gtaaaacggc ttgtcccgcg
     1621 tcatcggcgg gggtcataac gtgactccct taattctcat gtatctcctg acagcgaaat
     1681 gattgatgaa gaacaatggt ggatgaagaa caaagaagga gggagctttt gttcaagatg
     1741 aacaaagaac aatagtggat gaagaacaaa gtgaaaaaaa taaaaaaaaa tgtatggtta
     1801 aataaagagt aaagttacca ttgagactcc gtcaggagac tagagccaag ctgatctcct
     1861 ttgccccgga gatcaccatg gacgactttc tctatctcta cgatctagga agaaagttcg
     1921 acggagaagg tgacgatacc atgttcacca ccgataatga gaagattagc ctcttcaatt
     1981 tcagaaagaa tgctgaccca cagatggtta gagaggccta cgcggcaggt ctgatcaaga
     2041 cgatctaccc gagtaataat ctccaggaga tcaaatacct tcccaagaag gttaaagatg
     2101 cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata tttctcaaga
     2161 tcagaagtac tattccagta tggacgattc aaggcttgct tcataaacca aggcaagtaa
     2221 tagagattgg agtctctaag aaagtagttc ctactgaatc aaaggccatg gagtcaaaaa
     2281 ttcagatcga ggatctaaca gaactcgccg tgaagactgg cgaacagttc atacagagtc
     2341 ttttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac gacactctcg
     2401 tctactccaa gaatatcaaa gatacagtct cagaagacca aagggctatt gagacttttc
     2461 aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc tgtcacttca
     2521 tcaaaaggac agtagaaaag gaaggtggca cctacaaatg ccatcattgc gataaaggaa
     2581 aggctatcgt tcaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga
     2641 ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg
     2701 atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa gacccttcct
     2761 ctatataagg aagttcattt catttggaga ggactccggt atttttacaa caataccaca
     2821 acaaaacaaa caacaaacaa cattacaatt tactattcta gtcgaaatgg cctcctccga
     2881 gaacgtcatc accgagttca tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca
     2941 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggccaca acaccgtgaa
     3001 gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt
     3061 ccagtacggc tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct
     3121 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggc
     3181 gaccgtgacc caggactcct ccctgcagga cggctgcttc atctacaagg tgaagttcat
     3241 cggcgtgaac ttcccctccg acggccccgt gatgcagaag aagacgatgg gctgggaggc
     3301 ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagacac acaaggccct
     3361 gaagctgaag gacggcggcc actacctggt ggagttcaag tccatctaca tggccaagaa
     3421 gcccgtgcag ctgcccggct actactacgt ggacgccaag ctggacatca cctcccacaa
     3481 cgaggactac accatcgtgg agcagtacga gcgcaccgag ggccgccacc acctgttcct
     3541 gtaggcttcg gccatgctag agtccgcaaa aatcaccagt ctctctctac aaatctatct
     3601 ctctctattt ttctccagaa taatgtgtga gtagttccca gataagggaa ttagggttct
     3661 tatagggttt cgctcatgtg ttgagcatat aagaaaccct tagtatgtat ttgtatttgt
     3721 aaaatacttc tatcaataaa atttctaatt cctaaaacca aaatccagtg acctcgctgt
     3781 caggagtctc aatggtaact ttactcttta tttaaccata catttttttt tatttttttc
     3841 actttgttct tcatccacta ttgttctttg ttcatcttga acaaaagctc cctccttctt
     3901 tgttcttcat ccaccattgt tcttcatcaa tcatttcgct gtcaggagac gggacaagga
     3961 tgcgagctag cctgcaggaa ttgttgattt tgtgatgact gatggcagga tatatgcggt
     4021 tgtaattcat ttttattgtc taaatttctg tatttgtttg tttgttcggt tgtaaatttt
     4081 tttggaagaa caagaaaaga aaaaacaccc gttagggtgt ttttagttag tgtggcgcgc
     4141 cgacttgcga catgcggtcc tttgcaatta attattagaa aaattcatcc agcatcagat
     4201 gaaattgcag tttgttcata tccggattat caatgccata tttctgaaac agacgttttt
     4261 gcaggctcgg gctaaattcg cccaggcagt tccacagaat ggccagatcc tgataacgat
     4321 ccgcaatgcc cacacggccc acatcaatgc agccaatcag tttgccttca tcgaaaatca
     4381 ggttatccag gctaaaatcg ccgtgggtca ccacgctatc cgggctaaac ggcagcagtt
     4441 tatgcatttc tttccacacc tgttccaccg gccagccgtt acgttcatca tcaaaatcgc
     4501 tcgcatccac caggccgttg ttcatacggc tctgcgcctg ggccagacga aacacacgat
     4561 cgctgttaaa cgggcagttg cacaccggaa tgctatgcag acgacgcaga aacacggcca
     4621 gcgcatccac aatgttttcg ccgctatccg gatattcttc cagcacctga aacgcggttt
     4681 tgcccggaat cgcggtggtc agcagccacg catcatccgg ggtgcgaata aaatgtttaa
     4741 tggtcggcag cggcataaat tcggtcagcc agttcagacg caccatttca tcggtcacat
     4801 cgttcgccac gctgcctttg ccatgtttca gaaacagttc cggcgcatcc ggtttgccat
     4861 acagacgata aatggtcgcg ccgctctgac ccacgttatc acgcgcccat ttatagccat
     4921 acagatccgc atccatgttg ctgttcagac gcggacggct acagctcgtt tcacgctgaa
     4981 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     5041 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca agatcggatt
     5101 ggcggttatg cggttctacc ggcgcggcag cgttacccgt gtcggcggct ccaacggctc
     5161 gccatcgtcc agaaaacacg gctcatcggg catcggcagg cgctgctgcc cgcgccgttc
     5221 ccattcctcc gtttcggtca aggctggcag gtctggttcc atgcccggaa tgccgggctg
     5281 gctgggcggc tcctcgccgg ggccggtcgg tagttgctgc tcgcccggat acagggtcgg
     5341 gatgcggcgc aggtcgccat gccccaacag cgattcgtcc tggtcgtcgt gatcaaccac
     5401 cacggcggca ctgaacaccg acaggcgcaa ctggtcgcgg ggctggcccc acgccacgcg
     5461 gtcattgacc acgtaggccg acacggtgcc ggggccgttg agcttcacga cggagatcca
     5521 gcgctcggcc accaagtcct tgactgcgta ttggaccgtc cgcaaagaac gtccgatgag
     5581 cttggaaagt gtcttctggc tgaccaccac ggcgttctgg tggcccatct gcgccacgag
     5641 gtgatgcagc agcattgccg ccgtgggttt cctcgcaata agcccggccc acgcctcatg
     5701 cgctttgcgt tccgtttgca cccagtgacc gggcttgttc ttggcttgaa tgccgatttc
     5761 tctggactgc gtggccatgc ttatctccat gcggtagggg tgccgcacgg ttgcggcacc
     5821 atgcgcaatc agctgcaact tttcggcagc gcgacaacaa ttatgcgttg cgtaaaagtg
     5881 gcagtcaatt acagattttc tttaacctac gcaatgagct attgcggggg gtgccgcaat
     5941 gagctgttgc gtacccccct tttttaagtt gttgattttt aagtctttcg catttcgccc
     6001 tatatctagt tctttggtgc ccaaagaagg gcacccctgc ggggttcccc cacgccttcg
     6061 gcgcggctcc ccctccggca aaaagtggcc cctccggggc ttgttgatcg actgcgcggc
     6121 cttcggcctt gcccaaggtg gcgctgcccc cttggaaccc ccgcactcgc cgccgtgagg
     6181 ctcggggggc aggcgggcgg gcttcgccct tcgactgccc ccactcgcat aggcttgggt
     6241 cgttccaggc gcgtcaaggc caagccgctg cgcggtcgct gcgcgagcct tgacccgcct
     6301 tccacttggt gtccaaccgg caagcgaagc gcgcaggccg caggccggag gcttttcccc
     6361 agagaaaatt aaaaaaattg atggggcaag gccgcaggcc gcgcagttgg agccggtggg
     6421 tatgtggtcg aaggctgggt agccggtggg caatccctgt ggtcaagctc gtgggcaggc
     6481 gcagcctgtc catcagcttg tccagcaggg ttgtccacgg gccgagcgaa gcgagccagc
     6541 cggtggccgc tcgcggccat cgtccacata tccacgggct ggcaagggag cgcagcgacc
     6601 gcgcagggcg aagcccggag agcaagcccg taggggg
//
