LOCUS       40924_29321        5929 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-B2-PcrcmTFP1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5929)
  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 5929)
  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 5929)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5929)
  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..5929
                     /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"
     misc_feature    260..1289
                     /note="TAIR:AT4G28520; contains AtCRC promoter and 5' UTR"
     CDS             1290..1997
                     /label=mTFP1
                     /note="monomeric teal (cyan) fluorescent protein (Ai et
                     al., 2006)"
     CDS             1998..2012
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             2022..2045
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     misc_feature    2055..3232
                     /note="TAIR:AT4G28520; contains AtCRC 3' UTR and
                     terminator"
     misc_feature    3295..3318
                     /label=RB
                     /note="RB"
     regulatory      complement(3383..3423)
                     /regulatory_class="terminator"
     misc_feature    3433..3456
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(3470..4282)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    4384..4407
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(4411..5070)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(5071..5842)
                     /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 accgtctcat aacgaacgct catgctaagc tgcgcaaaat acttcctaat
      301 caaaacagta acaacgagta attagcaaaa tccgagcaga aaactctcac ccacctccga
      361 aattcacgtc ttcactaaaa ttttcgaaag gaatcgatca ataccaaccc attacacaaa
      421 atacataatc aaaatggcga gaatcgtacc tggaaacttt gcttcaagtc gcagagagag
      481 gaaaaggaag atcgtggaga aaggggttta gggtttaagc tcagacttct attggagtaa
      541 atgggacggt gtcacatttt ccgttttgga aatgaacttt gggctcacgt tatgggctat
      601 tagatatttg atgggctttc tagtaaatac aatataagtt attgggctta gtttaaataa
      661 gcccatgttg gaaatatttg acacatgtct tggctactag tgctaaacat gcaaccgaac
      721 agttgtcgag acaagtcgca gcatatacaa tggatcaaac acgcctagtg tcgccgcgtc
      781 tcgctcatgt gtcaccttgt ttcctcgttt ttttttaatt tttcataagt tcttttgttt
      841 tatcttcaat acaaattttt ggctgtatct tgcaaactct tcgatcatat cgccaatata
      901 cgtgaacact ggtgatctaa tttgttgtgt taattgttaa atttagattc tattctccgg
      961 tttaaaagtg aattatatgt atcatggtta aaacattgta agtaagatga taataaaatg
     1021 ataaatttag ttgatggata acgtgaagca aaaaatgaga tagatacatt tgattttgtc
     1081 gtattttgac atatgcggag agtgagctac gcgcatgaag atcaagagac acttgctcga
     1141 gctcacagag tgacgtgtaa aaagcttaga ctgaagtccc catgcaaacc taatcctacg
     1201 tggctcaaac cacgagctca cttgacaata tataaactcc tcctaagtcc cgttctcttc
     1261 atccatctct cacaacaaac aaaaagaaaa tggtttctaa aggtgaagag actacaatgg
     1321 gagtgatcaa gcctgatatg aagatcaagc tcaagatgga agggaacgtg aacggacacg
     1381 ctttcgttat tgaaggagaa ggggagggca agccttacga tggaactaac accatcaacc
     1441 ttgaagtgaa agagggtgct cctctccctt tctcttacga tatcctcact accgctttcg
     1501 cttacggaaa cagggctttc accaagtacc ctgatgatat ccctaactac ttcaagcagt
     1561 ctttcccaga gggatactct tgggagagaa ccatgacctt cgaggataag ggaatcgtga
     1621 aagtgaagtc tgatatctct atggaagagg attctttcat ctacgagatc catctcaagg
     1681 gcgagaactt ccctcctaac ggacctgtga tgcagaaaaa gactaccgga tgggatgctt
     1741 ctaccgagcg gatgtatgtg agagatggtg tgttgaaggg tgatgtgaag cacaagctgt
     1801 tgcttgaggg tggtggacat cacagagtgg atttcaagac catctacagg gctaagaagg
     1861 ctgttaagct ccctgattac cacttcgtgg atcatcgaat cgagatcctc aaccacgata
     1921 aggattacaa caaagtgacc gtgtacgagt ctgctgtggc tagaaactct accgatggag
     1981 gctctggtga ttacaaagac gatgacgata agggcagtgg agactataag gatgacgacg
     2041 ataaaggagc ataatgcact ggaggacaag gaaggatcca agtggtcaac gacaacggac
     2101 agaacgtgtt ggaccaacag gtgcagaagg gacagctcgt ggtcatccca caagggttcg
     2161 catacgttgt ccagtcccac ggaaacaagt tcgagtggat ctctttcaaa actaatgaaa
     2221 acgcaatgat cagcactttg gcgggtagaa cctcgctctt gagggcattg ccattggagg
     2281 tcatatcaaa tggtttccag atctctcccg aggaagctag gaagatcaag ttcaacacac
     2341 ttgagaccac tttgacccgc gctgccggta ggcaacaaca acagttgatc gaggagattg
     2401 tcgaggctta aatcaaaacg tttttctttt tcttaataaa gtatggtcag tttgtaatca
     2461 cgtcccttta cctttaacgt acgtgtaaaa tatgtgtctg cggcacctca cttgtaataa
     2521 cactttcttc tcataaataa aagggaagtt tcgagttaca tactataata tagcgccagt
     2581 tttttcgtct attccacaaa acataagttt gtgtccatct actgtagctt gagcttcgta
     2641 tatatacttg gtctttattc tttttttttt caaaatacgg ctttctgctt tctgtgggcc
     2701 taaacggagg cccaaacgaa ttaaagttcc gtcacttgga aacgttatcc tagattactc
     2761 ttgcggggaa aagtgtagat tagttccaat ttttgaagtg aaaaattgtg ttggattcta
     2821 tgtcgtaaca agaaatagca tggcctccaa agattattct ctctttcttt aggctttagc
     2881 ttctaaagct aagctacgga aaaactgtat cattcttctc ttttttttgg cttatgatcc
     2941 atataagaga gcttggaaga gctgcttttc atctcctgat cactagctca tacagttttt
     3001 gtaatttatt aacgaaacta tataaaaaaa gggaagccga aaaaacaaaa caaaaaacaa
     3061 ccttcatcgg ctgctcaaat gccaatcatc ccaattggtc tccattattg ttttctcgtt
     3121 actactcctc catctttgat ttaataattt tttgaaaaaa gttttactat atgaatgata
     3181 ttctctgtta gggaaatata aaaattgtaa gcaaatgatt ctatcgagct atgtcaggag
     3241 acgggacaag gatgcgagct agcctgcagg aattgttgat tttgtgatga ctgatggcag
     3301 gatatatgcg gttgtaattc atttttattg tctaaatttc tgtatttgtt tgtttgttcg
     3361 gttgtaaatt tttttggaag aacaagaaaa gaaaaaacac ccgttagggt gtttttagtt
     3421 agtgtggcgc gccgacttgc gacatgcggt cctttgcaat caactattag aaaaattcat
     3481 ccagcatcag atgaaattgc agtttgttca tatccggatt atcaatgcca tatttctgaa
     3541 acagacgttt ttgcaggctc gggctaaatt cgcccaggca gttccacaga atggccagat
     3601 cctgataacg atccgcaatg cccacacggc ccacatcaat gcagccaatc agtttgcctt
     3661 catcgaaaat caggttatcc aggctaaaat cgccgtgggt caccacgcta tccgggctaa
     3721 acggcagcag tttatgcatt tctttccaca cctgttccac cggccagccg ttacgttcat
     3781 catcaaaatc gctcgcatcc accaggccgt tgttcatacg gctctgcgcc tgggccagac
     3841 gaaacacacg atcgctgtta aacgggcagt tgcacaccgg aatgctatgc agacgacgca
     3901 gaaacacggc cagcgcatcc acaatgtttt cgccgctatc cggatattct tccagcacct
     3961 gaaacgcggt tttgcccgga atcgcggtgg tcagcagcca cgcatcatcc ggggtgcgaa
     4021 taaaatgttt aatggtcggc agcggcataa attcggtcag ccagttcaga cgcaccattt
     4081 catcggtcac atcgttcgcc acgctgcctt tgccatgttt cagaaacagt tccggcgcat
     4141 ccggtttgcc atacagacga taaatggtcg cgccgctctg acccacgtta tcacgcgccc
     4201 atttatagcc atacagatcc gcatccatgt tgctgttcag acgcggacgg ctacagctcg
     4261 tttcacgctg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     4321 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     4381 caagatcgga ttggcggtta tgcggttcta ccggcgcggc agcgttaccc gtgtcggcgg
     4441 ctccaacggc tcgccatcgt ccagaaaaca cggctcatcg ggcatcggca ggcgctgctg
     4501 cccgcgccgt tcccattcct ccgtttcggt caaggctggc aggtctggtt ccatgcccgg
     4561 aatgccgggc tggctgggcg gctcctcgcc ggggccggtc ggtagttgct gctcgcccgg
     4621 atacagggtc gggatgcggc gcaggtcgcc atgccccaac agcgattcgt cctggtcgtc
     4681 gtgatcaacc accacggcgg cactgaacac cgacaggcgc aactggtcgc ggggctggcc
     4741 ccacgccacg cggtcattga ccacgtaggc cgacacggtg ccggggccgt tgagcttcac
     4801 gacggagatc cagcgctcgg ccaccaagtc cttgactgcg tattggaccg tccgcaaaga
     4861 acgtccgatg agcttggaaa gtgtcttctg gctgaccacc acggcgttct ggtggcccat
     4921 ctgcgccacg aggtgatgca gcagcattgc cgccgtgggt ttcctcgcaa taagcccggc
     4981 ccacgcctca tgcgctttgc gttccgtttg cacccagtga ccgggcttgt tcttggcttg
     5041 aatgccgatt tctctggact gcgtggccat gcttatctcc atgcggtagg ggtgccgcac
     5101 ggttgcggca ccatgcgcaa tcagctgcaa cttttcggca gcgcgacaac aattatgcgt
     5161 tgcgtaaaag tggcagtcaa ttacagattt tctttaacct acgcaatgag ctattgcggg
     5221 gggtgccgca atgagctgtt gcgtaccccc cttttttaag ttgttgattt ttaagtcttt
     5281 cgcatttcgc cctatatcta gttctttggt gcccaaagaa gggcacccct gcggggttcc
     5341 cccacgcctt cggcgcggct ccccctccgg caaaaagtgg cccctccggg gcttgttgat
     5401 cgactgcgcg gccttcggcc ttgcccaagg tggcgctgcc cccttggaac ccccgcactc
     5461 gccgccgtga ggctcggggg gcaggcgggc gggcttcgcc cttcgactgc ccccactcgc
     5521 ataggcttgg gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc
     5581 cttgacccgc cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg
     5641 aggcttttcc ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt
     5701 ggagccggtg ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc
     5761 tcgtgggcag gcgcagcctg tccatcagct tgtccagcag ggttgtccac gggccgagcg
     5821 aagcgagcca gccggtggcc gctcgcggcc atcgtccaca tatccacggg ctggcaaggg
     5881 agcgcagcga ccgcgcaggg cgaagcccgg agagcaagcc cgtaggggg
//
