LOCUS       40924_35269        6389 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPLV28, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6389)
  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 6389)
  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 6389)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6389)
  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..6389
                     /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"
     regulatory      3898..5585
                     /note="pRPS5A; ribosomal protein S5A promoter"
                     /regulatory_class="promoter"
     misc_feature    5607..5636
                     /note="LIC; ligation-independent cloning (LIC) site"
     terminator      5662..5910
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5947..5965)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5986..6002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6010..6026)
                     /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(6034..6065)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6080..6101)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    6340..6364
                     /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 cggttgactc 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 gggtaccggg
     3901 ccataatcgt gagtagatat attactcaac ttttgattcg ctatttgcag tgcacctgtg
     3961 gcgttcatca catcttttgt gacactgttt gcactggtca ttgctattac aaaggacctt
     4021 cctgatgttg aaggagatcg aaagtaagta actgcacgca taaccatttt ctttccgctc
     4081 tttggctcaa tccatttgac agtcaaagac aatgtttaac cagctccgtt tgatatattg
     4141 tctttatgtg tttgttcaag catgtttagt taatcatgcc tttgattgat cttgaatagg
     4201 ttccaaatat caaccctggc aacaaaactt ggagtgagaa acattgcatt cctcggttct
     4261 ggacttctgc tagtaaatta tgtttcagcc atatcactag ctttctacat gcctcaggtg
     4321 aattcatcta tttccgtctt aactatttcg gttaattaaa gcacgaacac cattactgca
     4381 tgtagaagct tgataaacta tcgccaccaa tttatttttg ttgcgatatt gttactttcc
     4441 tcagtatgca gctttgaaaa gaccaaccct cttatccttt aacaatgaac aggtttttag
     4501 aggtagcttg atgattcctg cacatgtgat cttggcttca ggcttaattt tccaggtaaa
     4561 gcattatgag atactcttat atctcttaca tacttttgag ataatgcaca agaacttcat
     4621 aactatatgc tttagtttct gcatttgaca ctgccaaatt cattaatctc taatatcttt
     4681 gttgttgatc tttggtagac atgggtacta gaaaaagcaa actacaccaa ggtaaaatac
     4741 ttttgtacaa acataaactc gttatcacgg aacatcaatg gagtgtatat ctaacggagt
     4801 gtagaaacat ttgattattg caggaagcta tctcaggata ttatcggttt atatggaatc
     4861 tcttctacgc agagtatctg ttattcccct tcctctagct ttcaatttca tggtgaggat
     4921 atgcagtttt ctttgtatat cattcttctt cttctttgta gcttggagtc aaaatcggtt
     4981 ccttcatgta catacatcaa ggatatgtcc ttctgaattt ttatatcttg caataaaaat
     5041 gcttgtacca attgaaacac cagctttttg agttctatga tcactgactt ggttctaacc
     5101 aaaaaaaaaa aaatgtttaa tttacatatc taaaagtagg tttagggaaa cctaaacagt
     5161 aaaatatttg tatattattc gaatttcact catcataaaa acttaaattg caccataaaa
     5221 ttttgtttta ctattaatga tgtaatttgt gtaacttaag ataaaaataa tattccgtaa
     5281 gtttaccggc taaaaccacg tataaaccag ggaacctgtt aaaccggttc tttactggat
     5341 aaagaaatga aagcccatgt agacagctcc attagagccc aaaccctaaa tttctcatct
     5401 atataaaagg agtgacatta gggtttttgt tcgtcctctt aaagcttctc gttttctctg
     5461 ccgtctctct cattcgcgcg acgcaaacga tcttcaggtg atcttctttc tccaaatcct
     5521 ctctcataac tctgatttcg tacttgtgta tttgagctca cgctctgttt ctctcaccac
     5581 agccgggtac cgggcccccc ctcgagctag ttggaatagg ttaacccaac tccatactgc
     5641 agcccggggg atccgatccc cgatcgttca aacatttggc aataaagttt cttaagattg
     5701 aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat
     5761 gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc
     5821 ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa
     5881 ttatcgcgcg cggtgtcatc tatgtctaga gcggccgcca ccgcggtgga gctccagctt
     5941 ttgttccctt tagtgagggt taattccgag cttggcgtaa tcatggtcat agctgtttcc
     6001 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gacataaagt
     6061 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6121 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6181 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6241 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6301 cagaatcagg ggataacgca ggaaagaaca tgaaggcctt gacaggatat attggcgggt
     6361 aaactaagtc gctgtatgtg tttgtttga
//
