LOCUS       40924_31460        9021 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pMpGWB200 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9021)
  AUTHORS   Ishizaki K, Nishihama R, Ueda M, Inoue K, Ishida S, Nishimura Y, 
            Shikanai T, Kohchi T.
  TITLE     Development of Gateway Binary Vector Series with Four Different 
            Selection Markers for the Liverwort Marchantia polymorpha
  JOURNAL   PLoS ONE 10 (9), E0138876 (2015)
  PUBMED    26406247
REFERENCE   2  (bases 1 to 9021)
  AUTHORS   Nishihama R, Ishizaki K, Kohchi T.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-JUN-2015) Contact:Ryuichi Nishihama Kyoto University, 
            Graduate School of Biostudies; Kitashirakawa-oiwake-cho, Sakyo-ku, 
            Kyoto, Kyoto 606-8502, Japan URL :http://www.lif.kyoto-u.ac.jp/e/
REFERENCE   3  (bases 1 to 9021)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9021)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2015"; volume: "10"; issue: "9"; pages: "E0138876"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-JUN-2015) Contact:Ryuichi Nishihama Kyoto University, Graduate 
            School of Biostudies"; volume: " Kitashirakawa-oiwake-cho, Sakyo-ku,
            Kyoto, Kyoto 606-8502, Japan URL :http"; pages: 
            "//www.lif.kyoto-u.ac.jp/e"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(54..78)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     281..297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(301..357)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      362..614
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(648..664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(672..688)
                     /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(696..726)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(741..762)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     polyA_signal    complement(847..1019)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1079..1609)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MVRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(1672..2348)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     misc_feature    complement(2543..2567)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             3088..3876
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4125..4713
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4899..5039)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5383..5577)
                     /direction=LEFT
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(5646..6716)
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(7148..7774)
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
       61 cgccaatata tcctgtcaaa cactgatagt ttaaactgaa ggcgggaaac gacaatctga
      121 tccaagctca agctgctcta gcattcgcca ttcaggctgc gcaactgttg ggaagggcga
      181 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      241 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
      301 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaatttccc
      361 cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
      421 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
      481 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
      541 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
      601 tatgttacta gatcgggaat tggttccgga accaattcgt aatcatggtc atagctgttt
      661 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      721 tgtaaagcct ggggtgccta atgagtgagc taactcacat taggctgaat taggcgcgcc
      781 tatttctgaa ttcaacacat tgcggacgtt tttaatgtac tgaattaacg ccgaattaat
      841 tcgggggatc tggattttag tactggattt tggttttagg aattagattt tattgataga
      901 agtattttac aaatacaaat acatactaag ggtttcttat atgctcaaca catgagcgaa
      961 accctatagg aaccctaatt cccttatctg ggaactactc acacattatt atggagaaac
     1021 tcgagcttgt cgatcgactc taggccggga agccgatctc ggcttgaacg aattgttagg
     1081 tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc aactttgtat
     1141 agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa
     1201 gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc tgcctccggt
     1261 gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc tcaaacttgg
     1321 gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga
     1381 tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga tgttgggagt
     1441 aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc atggatttga
     1501 cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca cctaactttg
     1561 ttttagggcg actgccctgc tgcgtaacat cgttgctgct gcgtaccatg gtcgatcgac
     1621 agatctgcga aagctcgaga gagatagatt tgtagagaga gactggtgat ttcagcgtgt
     1681 cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag gatagtggga
     1741 ttgtgcgtca tcccttacgt cagtggagat atcacatcaa tccacttgct ttgaagacgt
     1801 ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc
     1861 actgtcggca gaggcatctt gaacgatagc ctttccttta tcgcaatgat ggcatttgta
     1921 ggtgccacct tccttttcta ctgtcctttt gatgaagtga cagatagctg ggcaatggaa
     1981 tccgaggagg tttcccgata ttaccctttg ttgaaaagtc tcaatagccc tttggtcttc
     2041 tgagactgta tctttgatat tcttggagta gacgagagtg tcgtgctcca ccatgttcac
     2101 atcaatccac ttgctttgaa gacgtggttg gaacgtcttc tttttccacg atgctcctcg
     2161 tgggtggggg tccatctttg ggaccactgt cggcagaggc atcttgaacg atagcctttc
     2221 ctttatcgca atgatggcat ttgtaggtgc caccttcctt ttctactgtc cttttgatga
     2281 agtgacagat agctgggcaa tggaatccga ggaggtttcc cgatattacc ctttgttgaa
     2341 aagtctcaat agccctttgg tcttctgaga ctgtatcttt gatattcttg gagtagacga
     2401 gagtgtcgtg ctccaccatg ttggcaagct gctctagcca atacgcaaac cgaattcaga
     2461 aataaattca gcctaattcg gcgttaattc agtacattaa aaacgtccgc aatgtgttat
     2521 taagttgtct aagcgtcaat ttgtttacac cacaatatat cctgccacca gccagccaac
     2581 agctccccga ccggcagctc ggcacaaaat caccactcga tacaggcagc ccatcagtcc
     2641 gggacggcgt cagcgggaga gccgttgtaa ggcggcagac tttgctcatg ttaccgatgc
     2701 tattcggaag aacggcaact aagctgccgg gtttgaaaca cggatgatct cgcggagggt
     2761 agcatgttga ttgtaacgat gacagagcgt tgctgcctgt gatcaattcg ggcacgaacc
     2821 cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc
     2881 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg
     2941 gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg
     3001 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc
     3061 agtcgcccta aaacaaagtt aaacatcatg ggggaagcgg tgatcgccga agtatcgact
     3121 caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta
     3181 catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg
     3241 gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg
     3301 gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt
     3361 gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa
     3421 tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg
     3481 gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag
     3541 gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg
     3601 ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc
     3661 cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg
     3721 gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat
     3781 cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac
     3841 tacgtgaaag gcgagatcac caaggtagtc ggcaaataat gtctagctag aaattcgttc
     3901 aagccgacgc cgcttcgcgg cgcggcttaa ctcaagcgtt agatgcacta agcacataat
     3961 tgctcacagc caaactatca ggtcaagtct gcttttatta tttttaagcg tgcataataa
     4021 gccctacaca aattgggaga tatatcatgc atgaccaaaa tcccttaacg tgagttttcg
     4081 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4141 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4201 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4261 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4321 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4381 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4441 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4501 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4561 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4621 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4681 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4741 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4801 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4861 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     4921 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     4981 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     5041 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5101 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     5161 caccgaaacg cgcgaggcag ggtgccttga tgtgggcgcc ggcggtcgag tggcgacggc
     5221 gcggcttgtc cgcgccctgg tagattgcct ggccgtaggc cagccatttt tgagcggcca
     5281 gcggccgcga taggccgacg cgaagcggcg gggcgtaggg agcgcagcga ccgaagggta
     5341 ggcgcttttt gcagctcttc ggctgtgcgc tggccagaca gttatgcaca ggccaggcgg
     5401 gttttaagag ttttaataag ttttaaagag ttttaggcgg aaaaatcgcc ttttttctct
     5461 tttatatcag tcacttacat gtgtgaccgg ttcccaatgt acggctttgg gttcccaatg
     5521 tacgggttcc ggttcccaat gtacggcttt gggttcccaa tgtacgtgct atccacagga
     5581 aagagacctt ttcgaccttt ttcccctgct agggcaattt gccctagcat ctgctccgta
     5641 cattaggaac cggcggatgc ttcgccctcg atcaggttgc ggtagcgcat gactaggatc
     5701 gggccagcct gccccgcctc ctccttcaaa tcgtactccg gcaggtcatt tgacccgatc
     5761 agcttgcgca cggtgaaaca gaacttcttg aactctccgg cgctgccact gcgttcgtag
     5821 atcgtcttga acaaccatct ggcttctgcc ttgcctgcgg cgcggcgtgc caggcggtag
     5881 agaaaacggc cgatgccggg atcgatcaaa aagtaatcgg ggtgaaccgt cagcacgtcc
     5941 gggttcttgc cttctgtgat ctcgcggtac atccaatcag ctagctcgat ctcgatgtac
     6001 tccggccgcc cggtttcgct ctttacgatc ttgtagcggc taatcaaggc ttcaccctcg
     6061 gataccgtca ccaggcggcc gttcttggcc ttcttcgtac gctgcatggc aacgtgcgtg
     6121 gtgtttaacc gaatgcaggt ttctaccagg tcgtctttct gctttccgcc atcggctcgc
     6181 cggcagaact tgagtacgtc cgcaacgtgt ggacggaaca cgcggccggg cttgtctccc
     6241 ttcccttccc ggtatcggtt catggattcg gttagatggg aaaccgccat cagtaccagg
     6301 tcgtaatccc acacactggc catgccggcc ggccctgcgg aaacctctac gtgcccgtct
     6361 ggaagctcgt agcggatcac ctcgccagct cgtcggtcac gcttcgacag acggaaaacg
     6421 gccacgtcca tgatgctgcg actatcgcgg gtgcccacgt catagagcat cggaacgaaa
     6481 aaatctggtt gctcgtcgcc cttgggcggc ttcctaatcg acggcgcacc ggctgccggc
     6541 ggttgccggg attctttgcg gattcgatca gcggccgctt gccacgattc accggggcgt
     6601 gcttctgcct cgatgcgttg ccgctgggcg gcctgcgcgg ccttcaactt ctccaccagg
     6661 tcatcaccca gcgccgcgcc gatttgtacc gggccggatg gtttgcgacc gctcacgccg
     6721 attcctcggg cttgggggtt ccagtgccat tgcagggccg gcagacaacc cagccgctta
     6781 cgcctggcca accgcccgtt cctccacaca tggggcattc cacggcgtcg gtgcctggtt
     6841 gttcttgatt ttccatgccg cctcctttag ccgctaaaat tcatctactc atttattcat
     6901 ttgctcattt actctggtag ctgcgcgatg tattcagata gcagctcggt aatggtcttg
     6961 ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct ccgccggcaa ctgaaagttg
     7021 acccgcttca tggctggcgt gtctgccagg ctggccaacg ttgcagcctt gctgctgcgt
     7081 gcgctcggac ggccggcact tagcgtgttt gtgcttttgc tcattttctc tttacctcat
     7141 taactcaaat gagttttgat ttaatttcag cggccagcgc ctggacctcg cgggcagcgt
     7201 cgccctcggg ttctgattca agaacggttg tgccggcggc ggcagtgcct gggtagctca
     7261 cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta cccggccagc gcctcggcaa
     7321 cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac gacaaaggcc gcttgtagcc
     7381 ttccatccgt gacctcaatg cgctgcttaa ccagctccac caggtcggcg gtggcccata
     7441 tgtcgtaagg gcttggctgc accggaatca gcacgaagtc ggctgccttg atcgcggaca
     7501 cagccaagtc cgccgcctgg ggcgctccgt cgatcactac gaagtcgcgc cggccgatgg
     7561 ccttcacgtc gcggtcaatc gtcgggcggt cgatgccgac aacggttagc ggttgatctt
     7621 cccgcacggc cgcccaatcg cgggcactgc cctggggatc ggaatcgact aacagaacat
     7681 cggccccggc gagttgcagg gcgcgggcta gatgggttgc gatggtcgtc ttgcctgacc
     7741 cgcctttctg gttaagtaca gcgataacct tcatgcgttc cccttgcgta tttgtttatt
     7801 tactcatcgc atcatatacg cagcgaccgc atgacgcaag ctgttttact caaatacaca
     7861 tcaccttttt agacggcggc gctcggtttc ttcagcggcc aagctggccg gccaggccgc
     7921 cagcttggca tcagacaaac cggccaggat ttcatgcagc cgcacggttg agacgtgcgc
     7981 gggcggctcg aacacgtacc cggccgcgat catctccgcc tcgatctctt cggtaatgaa
     8041 aaacggttcg tcctggccgt cctggtgcgg tttcatgctt gttcctcttg gcgttcattc
     8101 tcggcggccg ccagggcgtc ggcctcggtc aatgcgtcct cacggaaggc accgcgccgc
     8161 ctggcctcgg tgggcgtcac ttcctcgctg cgctcaagtg cgcggtacag ggtcgagcga
     8221 tgcacgccaa gcagtgcagc cgcctctttc acggtgcggc cttcctggtc gatcagctcg
     8281 cgggcgtgcg cgatctgtgc cggggtgagg gtagggcggg ggccaaactt cacgcctcgg
     8341 gccttggcgg cctcgcgccc gctccgggtg cggtcgatga ttagggaacg ctcgaactcg
     8401 gcaatgccgg cgaacacggt caacaccatg cggccggccg gcgtggtggt gtcggcccac
     8461 ggctctgcca ggctacgcag gcccgcgccg gcctcctgga tgcgctcggc aatgtccagt
     8521 aggtcgcggg tgctgcgggc caggcggtct agcctggtca ctgtcacaac gtcgccaggg
     8581 cgtaggtggt caagcatcct ggccagctcc gggcggtcgc gcctggtgcc ggtgatcttc
     8641 tcggaaaaca gcttggtgca gccggccgcg tgcagttcgg cccgttggtt ggtcaagtcc
     8701 tggtcgtcgg tgctgacgcg ggcatagccc agcaggccag cggcggcgct cttgttcatg
     8761 gcgtaatgtc tccggttcta gtcgcaagta ttctacttta tgcgactaaa acacgcgaca
     8821 agaaaacgcc aggaaaaggg cagggcggca gcctgtcgcg taacttagga cttgtgcgac
     8881 atgtcgtttt cagaagacgg ctgcactgaa cgtcagaagc cgactgcact atagcagcgg
     8941 aggggttgga tcaaagtact ttgatcccga ggggaaccct gtggttggca tgcacataca
     9001 aatggacgaa cggataaacc t
//
