LOCUS       40924_35284        9507 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPLV31, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9507)
  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 9507)
  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 9507)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9507)
  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..9507
                     /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)"
     promoter        2585..2930
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             2950..3972
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     regulatory      3976..4197
                     /note="t35S; CaMV 35S terminator"
                     /regulatory_class="terminator"
     polyA_signal    4206..4382
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     4530..4546
                     /note="M13F; M13 forward primer site"
     primer_bind     4550..4566
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4573..4591
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4606..8718
                     /note="pMP; MONOPTEROS (ARF5) promoter"
                     /regulatory_class="promoter"
     misc_feature    8725..8754
                     /note="LIC; ligation-independent cloning (LIC) site"
     terminator      8780..9028
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(9065..9083)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9104..9120)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(9128..9144)
                     /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(9152..9183)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(9198..9219)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    9458..9482
                     /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 gtacccctac tccaaaaatg tcaaagatac agtctcagaa gaccaaaggg
     2581 ctattgagac ttttcaacaa agggtaattt cgggaaacct cctcggattc cattgcccag
     2641 ctatctgtca cttcatcgaa aggacagtag aaaaggaagg tggctcctac aaatgccatc
     2701 attgcgataa aggaaaggct atcattcaag atgcctctgc cgacagtggt cccaaagatg
     2761 gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc
     2821 aagtggattg atgtgacatc tccactgacg taagggatga cgcacaatcc cactatcctt
     2881 cgcaagaccc ttcctctata taaggaagtt catttcattt ggagaggaca gcccaagctg
     2941 atccccccta tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     3001 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     3061 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     3121 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     3181 cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     3241 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttcttcagcc ggtcgcggag
     3301 gctatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     3361 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     3421 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     3481 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     3541 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     3601 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     3661 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     3721 ggatcgccac gcctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     3781 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     3841 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     3901 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     3961 agggcaaagg aatagagtag atgccgaccg aacaagagct gatttcgaga acgcctcagc
     4021 cagcaactcg cgcgagccta gcaaggcaaa tgcgagagaa cggccttacg cttggtggca
     4081 cagttctcgt ccacagttcg ctaagctcgc tcggctggtc gcgggagggc cggtcgcagt
     4141 gattcaggaa ttaattcggt acgctgaaat caccagtctc tctctacaaa tctatctctc
     4201 tctattttct ccataaataa tgtgtgagta gtttcccgat aagggaaatt agggttctta
     4261 tagggtttcg ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa
     4321 aatacttcta tcaataaaat ttctaattcc taaaaccaaa atccagtact aaaatccaga
     4381 tcgataacat taacgtttac aatttcgcgc cattcgccat tcaggctgcg caactgttgg
     4441 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     4501 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     4561 gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccgacgt gtgtgaatta
     4621 ccaaggcgtt gaacgacgtg aaagagcgcg tgcggctcga ttcttacgtc gttgaatatt
     4681 gaccgtcgga tcaaaactca tcgatccaca tagatgcgtg cgcgaaggca catgccgtta
     4741 tgaagggttc tcgatgaaca tcacgtgtcc tctagggatt ggctattatt gcaaactccg
     4801 ttttatgagg caaaaagaca atttaacctt tttaattttt atctatttgg taataaaaac
     4861 atttttattt cgtttgttta ttaattaatt aatgaggtta ttaaaggtgc cttagcaata
     4921 gcaggtaggc acttaattta atagtccaaa ctccaaagct aattaattta aaggacacaa
     4981 atcctaataa accccaaaca gttatttgcg accttttctt atcgaaatcc gttctttgaa
     5041 ttgtttactt tttgatcgat tttaataaca agattcacaa acacatataa gtgatttttt
     5101 aaaaaatgga atgagaaaat aaagaaaaga atttgaacgg ttaagaatat gataatcacg
     5161 aagatggcat gatattatag ttggagtggg ttacttggcg aaagaatcaa atagaatttt
     5221 cataaatttg gaatgtgacg tgtacttatt tctccaagcg ttacctgatt ttcgtattat
     5281 tattacggat tcgtgatctt cgtatcccat atggtcaatc gtttctattt cacttgtaaa
     5341 ttacaatatc tcgttcactg ttctataaca tactattacg tgctaagtat tactattgca
     5401 cagatcatct tgaacggtaa tcgacactta cccgggttaa tcagtattat tactttcatt
     5461 attctaacca aaaattaggt tgcacttgcc catgtgtacg acaccaatta ccttcaaaaa
     5521 gtttcttcaa agtatcttca tcaacttaac gtaaaatttt aaaccaacat aaatgttttt
     5581 ggttagccaa aagttagtta aaatttcact acgtattata catttttagg ttgctaatca
     5641 cttattgttc cactacattt ttattctttc caattattct acatgttata tctccatttt
     5701 acattacttg gtccactgtc actgttaaaa gccaccacta ttttatgttt ttttttcaat
     5761 tactacgatt tataagattt gtactatatt atgttacacc aatatggaag tatacttttt
     5821 gtgatgattt aagaaagtaa atctagtggc agtaagtaaa attataatga aactgttttc
     5881 ttatgggatt aataggaatc ggtctttgct ttgttagttt ggcaggagag agaggtaggt
     5941 aaggatgtga actgtgaagc gcagaaacgg cgccgtgtcg gtccaaaaat cattttcaaa
     6001 ataacattaa aatacacgtg tcgagtatgc taatgtacgc ctcctcttcc tctcttactc
     6061 aaccattgat ttgccgtcca cttcacgcgt gcctagcggt caaaattttc agacgcaaaa
     6121 gcaccaaata atgcttttca ctttcttttt ttctttcttc atttcttatt cttgtttttt
     6181 tacaagtttc caatctacat aagacaaact aacgtttagg aaggattcgg attgataatc
     6241 tgcatacact tgattttgat tggtatcctt ttctttgcat gtggttaaaa tttggtatgg
     6301 taagagtttt tatttttatg aatttaggat tattgaaggc attgtcacaa atgatataca
     6361 gtagagaaaa aaaaaaacga tacaagagaa aacaattgat aagaatgata aaattatatc
     6421 tagtaaacct catcgtggga tgttattcct cgttatacgt aatgactatt tgttccaaag
     6481 attttgctga tggcacacaa tcttggcacc ggaaatgaga gtttttccat tatcatcaaa
     6541 agcatgtttc actccttgct aaatttgaac tgtatattaa aatgtgctgc caacagcaaa
     6601 ttccacacat tatatacttg attggagatc ctttgattca aatatatctt agtgacaaac
     6661 gcgtatccga ataagtagtt atgtttatat agatattcta tcataatgat gctactatat
     6721 atccacttat aaattataat atgtttgatt atttttctct aatatgtgat aatgtggatt
     6781 gtgtactaat cctatatata acttaactat gctcatttgt attttgttta tgagtaatat
     6841 tacatgaagg aaagcgattt ttatttacgt atgattgata agtaatgata tggaaagtga
     6901 tatggtttca tatcatacaa tataatacaa tgaccgcatg tcaattattt cgtcgcaaga
     6961 aaacaatgga aaggtcaaac tagagatgtg tgcgtaaaaa aaaaaaatgt atgtaactaa
     7021 aaaggtctaa agatgtaatg gtacacacta tacacaatta gacataatta cacatgtggt
     7081 ttagagtatt agatattaat aatgagcaga acaagctatg tttacaggtg cagagaacca
     7141 agatattcct ttcttttggt tgttgtagat acaaatatat tttacaaagc tcaatttcaa
     7201 tacaaatgca agaaccacaa acaacagtcc tttcaattta tttggaggtg gtcttaactc
     7261 ttaagtatat ataaacaata ccaccttata acttttaagt tttaactaaa ccatacttaa
     7321 atctttttta atctctatac caaatcttgc aaatatcaaa ataacttttt ctaaccaaag
     7381 aaagaaaaca tttttgaagt ccttaacttg gaaaggaaaa gaacacccgg gcaaaaaaga
     7441 ttggaagata cacttcctcc cgtaacctct ttaggcttcg tgctaatata cgctattttg
     7501 acattctctc ctatcatacg caccttgtat tagagatctc ccaaagtctc accactcaca
     7561 tcccttttac gaaatctcag caagaaaaga ttaacatttt gagaaaagta tatattattg
     7621 caacgaaaaa agaagcctcc tcctttgtca taaagttctt aatcaatttt aatttattgc
     7681 cataatcaaa aaacatttta tattttacaa agttcttgaa aaagtaggac actttttaaa
     7741 ataatattca ctcatggtac ttattttaag ctttattatc tgccttataa cataagcatt
     7801 gaaagtactt gtacattcaa ctacgtttct gtgcgtatca gagattacgt gtgcttactc
     7861 tactttatta aatttttgac gtaaactgtt ctaaaacatt aaataataat atagatgtat
     7921 ctattgaaat accaaaactt tattttataa ttaatgggaa acaataaaat ataatattat
     7981 tcgtttaaag agtctaaagc tgcaacggca tcgccatata cagaaagttt taagcgcagg
     8041 ataagagcat gcacactctt tcccatttcc aagcaaaata aaacctatac atacacattt
     8101 atactaatac tcatatacat tagagagaga agaagaaaaa atgtgggatt tttcagagaa
     8161 ataattaaaa gaaagcaaaa gttttttttt tcttaaaagc ttttgtctct ctctcaattg
     8221 cttcttgttc ttgtttgcat ccggaaccaa agaaagaaac tgtgtggttt catgatggca
     8281 cctttggttt ctcttcttca ttgaagggtc gaatttggtc tgagaaaaga agagtggaga
     8341 gagagaaaga gaagaggcaa gagccccaca agagagacat gcttgtctgt gtagtcccta
     8401 ttttgtatgt tctttaatct ctcccactct gtttttgtca tgtgtaaggg ttttgtcctt
     8461 cgattttgga gtcttcttta accaaaaaaa aacagattcc atgaaatggc tttgtcttta
     8521 attcctctgg tttgtcctgg atttatcttc ttgtagcctc tttgctcaat agagctcttt
     8581 ttgtctcttc ttgtattaaa taaaataaaa tgaagactta gtaaagaaaa agaaaaactc
     8641 ttgagtcttg ctaaagccta gttagtgttg agtgtgggag agacaaaaca gaacattgaa
     8701 aaatctctct gtatgatgct cgagctagtt ggaataggtt aacccaactc catactgcag
     8761 cccgggggat ccgatccccg atcgttcaaa catttggcaa taaagtttct taagattgaa
     8821 tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt
     8881 aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc
     8941 gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt
     9001 atcgcgcgcg gtgtcatcta tgtctagagc ggccgccacc gcggtggagc tccagctttt
     9061 gttcccttta gtgagggtta attccgagct tggcgtaatc atggtcatag ctgtttcctg
     9121 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaaga cataaagtgt
     9181 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     9241 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     9301 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     9361 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     9421 gaatcagggg ataacgcagg aaagaacatg aaggccttga caggatatat tggcgggtaa
     9481 actaagtcgc tgtatgtgtt tgtttga
//
