LOCUS       40924_29371        5363 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-B4-TagRFP-T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5363)
  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 5363)
  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 5363)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5363)
  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..5363
                     /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(3845..4504)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(4505..5276)
                     /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 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 tctaccggcg cggcagcgtt acccgtgtcg gcggctccaa cggctcgcca tcgtccagaa
     3901 aacacggctc atcgggcatc ggcaggcgct gctgcccgcg ccgttcccat tcctccgttt
     3961 cggtcaaggc tggcaggtct ggttccatgc ccggaatgcc gggctggctg ggcggctcct
     4021 cgccggggcc ggtcggtagt tgctgctcgc ccggatacag ggtcgggatg cggcgcaggt
     4081 cgccatgccc caacagcgat tcgtcctggt cgtcgtgatc aaccaccacg gcggcactga
     4141 acaccgacag gcgcaactgg tcgcggggct ggccccacgc cacgcggtca ttgaccacgt
     4201 aggccgacac ggtgccgggg ccgttgagct tcacgacgga gatccagcgc tcggccacca
     4261 agtccttgac tgcgtattgg accgtccgca aagaacgtcc gatgagcttg gaaagtgtct
     4321 tctggctgac caccacggcg ttctggtggc ccatctgcgc cacgaggtga tgcagcagca
     4381 ttgccgccgt gggtttcctc gcaataagcc cggcccacgc ctcatgcgct ttgcgttccg
     4441 tttgcaccca gtgaccgggc ttgttcttgg cttgaatgcc gatttctctg gactgcgtgg
     4501 ccatgcttat ctccatgcgg taggggtgcc gcacggttgc ggcaccatgc gcaatcagct
     4561 gcaacttttc ggcagcgcga caacaattat gcgttgcgta aaagtggcag tcaattacag
     4621 attttcttta acctacgcaa tgagctattg cggggggtgc cgcaatgagc tgttgcgtac
     4681 cccccttttt taagttgttg atttttaagt ctttcgcatt tcgccctata tctagttctt
     4741 tggtgcccaa agaagggcac ccctgcgggg ttcccccacg ccttcggcgc ggctccccct
     4801 ccggcaaaaa gtggcccctc cggggcttgt tgatcgactg cgcggccttc ggccttgccc
     4861 aaggtggcgc tgcccccttg gaacccccgc actcgccgcc gtgaggctcg gggggcaggc
     4921 gggcgggctt cgcccttcga ctgcccccac tcgcataggc ttgggtcgtt ccaggcgcgt
     4981 caaggccaag ccgctgcgcg gtcgctgcgc gagccttgac ccgccttcca cttggtgtcc
     5041 aaccggcaag cgaagcgcgc aggccgcagg ccggaggctt ttccccagag aaaattaaaa
     5101 aaattgatgg ggcaaggccg caggccgcgc agttggagcc ggtgggtatg tggtcgaagg
     5161 ctgggtagcc ggtgggcaat ccctgtggtc aagctcgtgg gcaggcgcag cctgtccatc
     5221 agcttgtcca gcagggttgt ccacgggccg agcgaagcga gccagccggt ggccgctcgc
     5281 ggccatcgtc cacatatcca cgggctggca agggagcgca gcgaccgcgc agggcgaagc
     5341 ccggagagca agcccgtagg ggg
//
