LOCUS       40924_35159        6892 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPLV12, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6892)
  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 6892)
  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 6892)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6892)
  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..6892
                     /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"
     primer_bind     4617..4633
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4654..4683
                     /note="LIC; ligation-independent cloning (LIC) site"
     CDS             4700..4720
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4727..6148
                     /label=tdTomato
                     /note="tandem dimeric (pseudo-monomeric) derivative of
                     DsRed (Shaner et al., 2004)"
     terminator      6165..6413
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(6450..6468)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6489..6505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6513..6529)
                     /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(6537..6568)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6583..6604)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    6843..6867
                     /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 gtaccgggcc ccccctcgag
     4621 gtcgacggta tcgataagct tgatatcgaa ttctagttgg aatgggttaa cccaactcca
     4681 taaggatccc atgactagtc ctaagaagaa gaggaaggtt ggaactagta agggcgagga
     4741 ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga
     4801 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     4861 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat
     4921 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc
     4981 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac
     5041 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg
     5101 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     5161 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     5221 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     5281 cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     5341 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctggg
     5401 gcatggcacc ggcagcaccg gcagcggcag ctccggcacc gcctcctccg aggacaacaa
     5461 catggccgtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg
     5521 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     5581 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     5641 gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa
     5701 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct
     5761 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat
     5821 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
     5881 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     5941 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
     6001 gaagcccgtg caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     6061 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     6121 cctgtacggc atggacgagc tgtacaagta aggatccgat ccccgatcgt tcaaacattt
     6181 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
     6241 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
     6301 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
     6361 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtct agagcggccg
     6421 ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattcc gagcttggcg
     6481 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6541 atacgagccg gaagacataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6601 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6661 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6721 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6781 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgaaggc
     6841 cttgacagga tatattggcg ggtaaactaa gtcgctgtat gtgtttgttt ga
//
