LOCUS       40924_29326        5007 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-B2-PetohmNEON, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5007)
  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 5007)
  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 5007)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5007)
  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..5007
                     /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"
     regulatory      260..449
                     /label=CaMV 35S
                     /note="CaMV 35S"
                     /regulatory_class="terminator"
     regulatory      450..804
                     /note="alcA; FMV 34S"
                     /regulatory_class="promoter"
     misc_feature    826..881
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             883..1590
                     /label=mNeonGreen
                     /note="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
     3'UTR           1758..1974
     terminator      2056..2308
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2373..2396
                     /label=RB
                     /note="RB"
     regulatory      complement(2461..2501)
                     /regulatory_class="terminator"
     misc_feature    2511..2534
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(2548..3360)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    3462..3485
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(3489..4148)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(4149..4920)
                     /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 accgtctcaa aatcaccagt ctctctctac aaatctatct ctctctataa
      301 taatgtgtga gtagttccca gataagggaa ttagggttct tatagggttt cgctcatgtg
      361 ttgagcatat aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa
      421 atttctaatt cctaaaacca aaatccagtg acccatgcgg agccgcattg gaggccatgc
      481 ggagccgcac gcgttaacaa gagcggctcc gcttgacctc tcgggatagt tccgacctag
      541 gattggatgc atgcggaacc gcacgagggc ggggcggaaa ttgacacacc actcctctcc
      601 acgcaccgtt caagaggtac gcgtatagag ccgtatagag cagagacgga gcactttctg
      661 gtactgtccg cacgggatgt ccgcacggag agccacaaac gagcggggcc ccgtacgtgc
      721 tctcctaccc caggatcgca tccccgcata gctgaacatc tatataagaa ggcattcatt
      781 cccatttgaa ggatcatcag atactgaacc aatattattt ttacaacaat taccaacaac
      841 aacaaacaac aaacaacatt acaattacta tttacaatta caatggtgag caagggagag
      901 gaggataaca tggcctctct cccagctaca catgagcttc acatctttgg atccatcaac
      961 ggtgtggact ttgacatggt gggtcaggga accggaaatc caaatgatgg atatgaggag
     1021 cttaacctta agtccaccaa gggtgacctc cagttctccc catggattct tgtccctcat
     1081 atcggatatg gattccatca gtaccttcct taccctgatg gaatgtctcc gtttcaagcc
     1141 gcaatggttg atggatccgg ataccaagtc catagaacaa tgcagtttga agatggtgcc
     1201 tcccttactg ttaactacag atacacctac gagggaagcc acatcaaagg agaggcccag
     1261 gtgaagggaa ctggtttccc tgctgacggt cctgtgatga ccaactctct taccgctgct
     1321 gactggtgca ggtctaagaa aacttaccct aacgacaaaa ccatcatcag tacctttaag
     1381 tggagttaca ccactggaaa tggcaagaga tacagaagca ctgctagaac cacctacacc
     1441 tttgccaagc caatggctgc taactatctt aagaaccagc ctatgtacgt gttccgtaag
     1501 actgagctca agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc
     1561 gatgtgatgg gaatggacga gctatacaaa taagtgaagc tcatatccag atgaaggcag
     1621 cagcattgag aaatgttcaa aatcgtttat ttggcttgga tggaaacgtc ggaacacaag
     1681 aagaggacac agagagacac accgctggtg atgttaatcg caacatgcac aacctcctcg
     1741 gtgtgagggg agtgtagtgg tctcggtatc tatcataaac tctacctggg tgagagtcta
     1801 atcatccagt tgtttttaga ttcctgttag catccttttc tccgctttaa tagcagtaca
     1861 ttcagtgagg ttttacctcc atatgttcta gtctgttatt gtcgaacaca ggcccttgta
     1921 tctgatgtag cgagtgcttc actccattcg ggttatagtt cttgtgcaag agacaaaaaa
     1981 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2041 aaatgcatgc ctgcagatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct
     2101 gttgccggtc ttgcgatgat tatcatctaa tttctgttga attacgttaa gcatgtaata
     2161 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa
     2221 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg
     2281 cgcgcggtgt catctatgtt actcgatctc gtcaggagac gggacaagga tgcgagctag
     2341 cctgcaggaa ttgttgattt tgtgatgact gatggcagga tatatgcggt tgtaattcat
     2401 ttttattgtc taaatttctg tatttgtttg tttgttcggt tgtaaatttt tttggaagaa
     2461 caagaaaaga aaaaacaccc gttagggtgt ttttagttag tgtggcgcgc cgacttgcga
     2521 catgcggtcc tttgcaatca actattagaa aaattcatcc agcatcagat gaaattgcag
     2581 tttgttcata tccggattat caatgccata tttctgaaac agacgttttt gcaggctcgg
     2641 gctaaattcg cccaggcagt tccacagaat ggccagatcc tgataacgat ccgcaatgcc
     2701 cacacggccc acatcaatgc agccaatcag tttgccttca tcgaaaatca ggttatccag
     2761 gctaaaatcg ccgtgggtca ccacgctatc cgggctaaac ggcagcagtt tatgcatttc
     2821 tttccacacc tgttccaccg gccagccgtt acgttcatca tcaaaatcgc tcgcatccac
     2881 caggccgttg ttcatacggc tctgcgcctg ggccagacga aacacacgat cgctgttaaa
     2941 cgggcagttg cacaccggaa tgctatgcag acgacgcaga aacacggcca gcgcatccac
     3001 aatgttttcg ccgctatccg gatattcttc cagcacctga aacgcggttt tgcccggaat
     3061 cgcggtggtc agcagccacg catcatccgg ggtgcgaata aaatgtttaa tggtcggcag
     3121 cggcataaat tcggtcagcc agttcagacg caccatttca tcggtcacat cgttcgccac
     3181 gctgcctttg ccatgtttca gaaacagttc cggcgcatcc ggtttgccat acagacgata
     3241 aatggtcgcg ccgctctgac ccacgttatc acgcgcccat ttatagccat acagatccgc
     3301 atccatgttg ctgttcagac gcggacggct acagctcgtt tcacgctgaa tatggctcat
     3361 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     3421 tttatcttgt gcaatgtaac atcagagatt ttgagacaca agatcggatt ggcggttatg
     3481 cggttctacc ggcgcggcag cgttacccgt gtcggcggct ccaacggctc gccatcgtcc
     3541 agaaaacacg gctcatcggg catcggcagg cgctgctgcc cgcgccgttc ccattcctcc
     3601 gtttcggtca aggctggcag gtctggttcc atgcccggaa tgccgggctg gctgggcggc
     3661 tcctcgccgg ggccggtcgg tagttgctgc tcgcccggat acagggtcgg gatgcggcgc
     3721 aggtcgccat gccccaacag cgattcgtcc tggtcgtcgt gatcaaccac cacggcggca
     3781 ctgaacaccg acaggcgcaa ctggtcgcgg ggctggcccc acgccacgcg gtcattgacc
     3841 acgtaggccg acacggtgcc ggggccgttg agcttcacga cggagatcca gcgctcggcc
     3901 accaagtcct tgactgcgta ttggaccgtc cgcaaagaac gtccgatgag cttggaaagt
     3961 gtcttctggc tgaccaccac ggcgttctgg tggcccatct gcgccacgag gtgatgcagc
     4021 agcattgccg ccgtgggttt cctcgcaata agcccggccc acgcctcatg cgctttgcgt
     4081 tccgtttgca cccagtgacc gggcttgttc ttggcttgaa tgccgatttc tctggactgc
     4141 gtggccatgc ttatctccat gcggtagggg tgccgcacgg ttgcggcacc atgcgcaatc
     4201 agctgcaact tttcggcagc gcgacaacaa ttatgcgttg cgtaaaagtg gcagtcaatt
     4261 acagattttc tttaacctac gcaatgagct attgcggggg gtgccgcaat gagctgttgc
     4321 gtacccccct tttttaagtt gttgattttt aagtctttcg catttcgccc tatatctagt
     4381 tctttggtgc ccaaagaagg gcacccctgc ggggttcccc cacgccttcg gcgcggctcc
     4441 ccctccggca aaaagtggcc cctccggggc ttgttgatcg actgcgcggc cttcggcctt
     4501 gcccaaggtg gcgctgcccc cttggaaccc ccgcactcgc cgccgtgagg ctcggggggc
     4561 aggcgggcgg gcttcgccct tcgactgccc ccactcgcat aggcttgggt cgttccaggc
     4621 gcgtcaaggc caagccgctg cgcggtcgct gcgcgagcct tgacccgcct tccacttggt
     4681 gtccaaccgg caagcgaagc gcgcaggccg caggccggag gcttttcccc agagaaaatt
     4741 aaaaaaattg atggggcaag gccgcaggcc gcgcagttgg agccggtggg tatgtggtcg
     4801 aaggctgggt agccggtggg caatccctgt ggtcaagctc gtgggcaggc gcagcctgtc
     4861 catcagcttg tccagcaggg ttgtccacgg gccgagcgaa gcgagccagc cggtggccgc
     4921 tcgcggccat cgtccacata tccacgggct ggcaagggag cgcagcgacc gcgcagggcg
     4981 aagcccggag agcaagcccg taggggg
//
