LOCUS       40924_35089        5016 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPLV03_v2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5016)
  AUTHORS   Wendrich JR, Liao CY, van den Berg WA, De Rybel B, Weijers D.
  TITLE     Ligation-independent cloning for plant research
  JOURNAL   Methods Mol. Biol. 1284, 421-431 (2015)
  PUBMED    25757785
REFERENCE   2  (bases 1 to 5016)
  AUTHORS   Wendrich JR, Liao C-Y., van den Berg WAM., De Rybel B, Weijers D.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2014) Laboratory of Biochemistry, Wageningen 
            University, Dreijenlaan 3, Wageningen 6703HA, The Netherlands
REFERENCE   3  (bases 1 to 5016)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5016)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods 
            Mol. Biol. 1284, 421-431 (2015)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-JUN-2014) Laboratory of Biochemistry, Wageningen University, 
            Dreijenlaan 3, Wageningen 6703HA, The Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     45..61
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    82..111
                     /note="LIC; ligation-independent cloning (LIC) site"
     terminator      125..373
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    395..419
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(510..1098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1272..2084)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      2375..2810
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    2945..2967
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     promoter        3029..3374
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3394..4416
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     polyA_signal    4650..4826
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     4994..5010
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt cgacggtatc
       61 gataagcttg atatcgaatt ctagttggaa tgggttaacc caactccata aggatccgat
      121 ccccgatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct
      181 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta
      241 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta
      301 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc
      361 atctatgtct agagcggccg ccacgcgtag gccttgacag gatatattgg cgggtaaact
      421 aagtcgctgt atgtgtttgt ttgagatctc atgtgagcaa aaggccagca aaaggccagg
      481 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      541 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      601 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      661 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      721 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
      781 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      841 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
      901 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
      961 ggtatctgcg ctctgctgaa gccagttacc ttcggaagaa gagttggtag ctcttgatcc
     1021 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1081 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1141 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1201 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatgt gtaacattgg
     1261 tctagtgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     1321 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     1381 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     1441 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     1501 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     1561 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     1621 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     1681 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     1741 tcaggatatt cttctaatac ctggaatgct gttttccctg ggatcgcagt ggtgagtaac
     1801 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     1861 agccagttta gtctgaccat ctcatctgta acaacattgg caacgctacc tttgccatgt
     1921 ttcagaaaca actctggcgc atcgggcttc ccatacaatc ggtagattgt cgcacctgat
     1981 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     2041 aatcgcggcc ttgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     2101 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     2161 taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag
     2221 ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag
     2281 ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact
     2341 ttctggctgg atgatggggc gattcaggcg atccccatcc aacagcccgc cgtcgagcgg
     2401 gcttttttat ccccggaagc ctgtggatag agggtagtta tccacgtgaa accgctaatg
     2461 ccccgcaaag ccttgattca cggggctttc cggcccgctc caaaaactat ccacgtgaaa
     2521 tcgctaatca gggtacgtga aatcgctaat cggagtacgt gaaatcgcta ataaggtcac
     2581 gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata gccctttcag atcaacagct
     2641 tgcaaacacc cctcgctccg gcaagtagtt acagcaagta gtatgttcaa ttagcttttc
     2701 aattatgaat atatatatca attattggtc gcccttggct tgtggacaat gcgctacgcg
     2761 caccggctcc gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc cagcgccttt
     2821 gcccacaacc cggcggccgg ccgcaacaga tcgttttata aatttttttt tttgaaaaag
     2881 aaaaagcccg aaaggcggca acctctcggg cttctggatt tccgatcccc ggaattagag
     2941 atcttggcag gatatattgt ggtgtaacgt tatcgtaccc ctactccaaa aatgtcaaag
     3001 atacagtctc agaagaccaa agggctattg agacttttca acaaagggta atttcgggaa
     3061 acctcctcgg attccattgc ccagctatct gtcacttcat cgaaaggaca gtagaaaagg
     3121 aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggctatcatt caagatgcct
     3181 ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaagaag
     3241 acgttccaac cacgtcttca aagcaagtgg attgatgtga catctccact gacgtaaggg
     3301 atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga agttcatttc
     3361 atttggagag gacagcccaa gctgatcccc cctatgaaaa agcctgaact caccgcgacg
     3421 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     3481 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     3541 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     3601 gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag cctgacctat
     3661 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     3721 gctgttcttc agccggtcgc ggaggctatg gatgcgatcg ctgcggccga tcttagccag
     3781 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     3841 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     3901 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     3961 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     4021 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     4081 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     4141 gagcggaggc atccggagct tgcaggatcg ccacgcctcc gggcgtatat gctccgcatt
     4201 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     4261 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     4321 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     4381 aaccgacgcc ccagcactcg tccgagggca aaggaataga gtagatgccg accgaacaag
     4441 agctgatttc gagaacgcct cagccagcaa ctcgcgcgag cctagcaagg caaatgcgag
     4501 agaacggcct tacgcttggt ggcacagttc tcgtccacag ttcgctaagc tcgctcggct
     4561 ggtcgcggga gggccggtcg cagtgattca ggaattaatt cggtacgctg aaatcaccag
     4621 tctctctcta caaatctatc tctctctatt ttctccataa ataatgtgtg agtagtttcc
     4681 cgataaggga aattagggtt cttatagggt ttcgctcatg tgttgagcat ataagaaacc
     4741 cttagtatgt atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac
     4801 caaaatccag tactaaaatc cagatcgata acattaacgt ttacaatttc gcgccattcg
     4861 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4921 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     4981 cagtcacgac gttgtaaaac gacggccagt gaattg
//
