LOCUS       40924_35254        6042 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPLV25, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6042)
  AUTHORS   De Rybel B, van den Berg W, Lokerse A, Liao CY, van Mourik H, Moller
            B, Peris CL, Weijers D.
  TITLE     A versatile set of ligation-independent cloning vectors for 
            functional studies in plants
  JOURNAL   Plant Physiol. 156 (3), 1292-1299 (2011)
  PUBMED    21562332
REFERENCE   2  (bases 1 to 6042)
  AUTHORS   De Rybel B, van den Berg W, Weijers D.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2011) Laboratory of Biochemistry, Wageningen 
            University, Dreijenlaan 3, Wageningen 6703HA, The Netherlands
REFERENCE   3  (bases 1 to 6042)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6042)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Physiol."; date: "2011"; volume: "156"; issue: "3"; pages: 
            "1292-1299"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-MAY-2011) Laboratory of Biochemistry, Wageningen University, 
            Dreijenlaan 3, Wageningen 6703HA, The Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(66..654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(828..1640)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1931..2366
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    2501..2523
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     terminator      complement(2534..2786)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(2823..3371)
                     /label=BlpR
                     /note="phosphinothricin acetyltransferase"
     promoter        complement(3413..3592)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     3819..3835
                     /note="M13F; M13 forward primer site"
     primer_bind     3839..3855
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3865..3883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4897..5242
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    5260..5289
                     /note="LIC; ligation-independent cloning (LIC) site"
     terminator      5315..5563
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5600..5618)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5639..5655)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5663..5679)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5687..5718)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5733..5754)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    5993..6017
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gatctcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
       61 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      541 gttaccttcg gaagaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      721 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      781 tttaaatcaa tctaaagtat atatgtgtaa cattggtcta gtgattagaa aaactcatcg
      841 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
      901 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
      961 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     1021 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     1081 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     1141 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     1201 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     1261 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     1321 aatgctgttt tccctgggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     1381 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     1441 tctgtaacaa cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     1501 ggcttcccat acaatcggta gattgtcgca cctgattgcc cgacattatc gcgagcccat
     1561 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggccttga gcaagacgtt
     1621 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     1681 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     1741 acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt
     1801 cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc
     1861 tacaacaaag ctctcatcaa ccgtggctcc ctcactttct ggctggatga tggggcgatt
     1921 caggcgatcc ccatccaaca gcccgccgtc gagcgggctt ttttatcccc ggaagcctgt
     1981 ggatagaggg tagttatcca cgtgaaaccg ctaatgcccc gcaaagcctt gattcacggg
     2041 gctttccggc ccgctccaaa aactatccac gtgaaatcgc taatcagggt acgtgaaatc
     2101 gctaatcgga gtacgtgaaa tcgctaataa ggtcacgtga aatcgctaat caaaaaggca
     2161 cgtgagaacg ctaatagccc tttcagatca acagcttgca aacacccctc gctccggcaa
     2221 gtagttacag caagtagtat gttcaattag cttttcaatt atgaatatat atatcaatta
     2281 ttggtcgccc ttggcttgtg gacaatgcgc tacgcgcacc ggctccgccc gtggacaacc
     2341 gcaagcggtt gcccaccgtc gagcgccagc gcctttgccc acaacccggc ggccggccgc
     2401 aacagatcgt tttataaatt tttttttttg aaaaagaaaa agcccgaaag gcggcaacct
     2461 ctcgggcttc tggatttccg atccccggaa ttagagatct tggcaggata tattgtggtg
     2521 taacgttatc cccgatctag taacatagat gacaccgcgc gcgataattt atcctagttt
     2581 gcgcgctata ttttgttttc tatcgcgtat taaatgtata attgcgggac tctaatcata
     2641 aaaacccatc tcataaataa cgtcatgcat tacatgttaa ttattacatg cttaacgtaa
     2701 ttcaacagaa attatatgat aatcatcgca agaccggcaa caggattcaa tcttaagaaa
     2761 ctttattgcc aaatgtttga acgatcgagc ttgcatgctt gcagagatcc gccgggtcat
     2821 cagatttcgg tgacgggcag gaccggacgg ggcggcaccg gcaggctgaa gtccagctgc
     2881 cagaaaccca cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccgcgg
     2941 ggggcatatc cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca
     3001 gcgaccacgc tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg
     3061 gagcccagtc ccgtccgctg gtggcggggg gagacgtaca cggtcgactc ggccgtccag
     3121 tcgtaggcgt tgcgtgcctt ccagggaccc gcgtaggcga tgccggcgac ctcgccgtcc
     3181 acctcggcga cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc
     3241 tgcggttcct gcggctcggt acggaagttg accgtgcttg tctcgatgta gtggttgacg
     3301 atggtgcaga ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt
     3361 tctgggctca tggtagatcc cctcgatcga gttgagagtg aatatgagac tctaattgga
     3421 taccgagggg aatttatgga acgtcagtgg agcatttttg acaagaaata tttgctagct
     3481 gatagtgacc ttaggcgact tttgaacgcg caataatggt ttctgacgta tgtgcttagc
     3541 tcattaaact ccagaaaccc gcggctcagt ggctccttca acgttgcggt tctgtcagtt
     3601 ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt cataacgtga ctcccttaat
     3661 tctccgctca tgatcgataa cattaacgct tacaatttcc attcgccatt caggctgcgc
     3721 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     3781 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     3841 aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtaccatc
     3901 gacgaattaa ttccaatccc acaaaaatct gagcttaaca gcacagttgc tcctctcaga
     3961 gcagaatcgg gtattcaaca ccctcatatc aactactacg ttgtgtataa cggtccacat
     4021 gccggtatat acgatgactg gggttgtaca aaggcggcaa caaacggcgt tcccggagtt
     4081 gcacacaaga aatttgccac tattacagag gcaagagcag cagctgacgc gtacacaaca
     4141 agtcagcaaa caggcaggtt gaacttcatc cccaaaggag aagctcaact caagcccaag
     4201 agctttgcta aggccctaac aagcccacca aagcaaaaag cccactggct cacgctagga
     4261 accaaaaggc ccagcagtga tccagcccca aaagagatct cctttgcccc ggagattaca
     4321 atggacgatt tcctctatct ttacgatcta ggaaggaagt tcgaaggtga aggtgacgac
     4381 actatgttca ccactgataa tgagaaggtt agcctcttca atttcagaaa gaatgctgac
     4441 ccacagatgg ttagagaggc ctacgcagca ggtctcatca agacgatcta cccgagtaac
     4501 aatctccagg agatcaaata ccttcccaag aaggttaaag atgcagtcaa aagattcagg
     4561 actaattgca tcaagaacac agagaaagac atatttctca agatcagaag tactattcca
     4621 gtatggacga ttcaaggctt gcttcataaa ccaaggcaag taatagagat tggagtctct
     4681 aaaaaggtag ttcctactga atctaaggcc atgcatggag tctaagattc aaatcgagga
     4741 tctaacagaa ctcgccgtga agactggcga acagttcata cagagtcttt tacgactcaa
     4801 tgacaagaag aaaatcttcg tcaacatggt ggagcacgac actctggtct actccaaaaa
     4861 tgtcaaagat acagtctcag aagaccaaag ggctattgag acttttcaac aaaggataat
     4921 ttcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatcg aaaggacagt
     4981 agaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg ctatcattca
     5041 agatctctct gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga
     5101 aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgaca tctccactga
     5161 cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag
     5221 ttcatttcat ttggagagga cacggggccc cccctcgagc tagttggaat aggttaaccc
     5281 aactccatac tgcagcccgg gggatccgat ccccgatcgt tcaaacattt ggcaataaag
     5341 tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa
     5401 ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt
     5461 tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc
     5521 aaactaggat aaattatcgc gcgcggtgtc atctatgtct agagcggccg ccaccgcggt
     5581 ggagctccag cttttgttcc ctttagtgag ggttaattcc gagcttggcg taatcatggt
     5641 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5701 gaagacataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5761 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5821 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5881 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5941 atacggttat ccacagaatc aggggataac gcaggaaaga acatgaaggc cttgacagga
     6001 tatattggcg ggtaaactaa gtcgctgtat gtgtttgttt ga
//
