LOCUS       40924_33667        6624 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pnVGW3 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6624)
  AUTHORS   Nishimura K, Ishikawa S, Matsunami E, Yamauchi J, Homma K, Faulkner 
            C, Oparka K, Jisaka M, Nagaya T, Yokota K, Nakagawa T.
  TITLE     New Gateway-compatible vectors for a high-throughput protein-protein
            interaction analysis by a bimolecular fluorescence complementation 
            (BiFC) assay in plants and their application to a plant clathrin 
            structure analysis
  JOURNAL   Biosci. Biotechnol. Biochem. 79 (12), 1995-2006 (2015)
  PUBMED    26193449
REFERENCE   2  (bases 1 to 6624)
  AUTHORS   Nishimura K, Matsunami E.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-FEB-2015) Contact:Kohji Nishimura Shimane University, 
            Organization of Research; 1060 Nishikawatsu, Matsue, Shimane 
            690-8504, Japan
REFERENCE   3  (bases 1 to 6624)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6624)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biosci. 
            Biotechnol. Biochem."; date: "2015"; volume: "79"; issue: "12"; 
            pages: "1995-2006"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-FEB-2015) Contact:Kohji Nishimura Shimane University, 
            Organization of Research; 1060 Nishikawatsu, Matsue, Shimane 
            690-8504, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     constructed using pUC119.
FEATURES             Location/Qualifiers
     source          1..6624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        745..1090
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    1111..1740
                     /note="n-terminal fragment of yellow fluorescent protein 
                     (replacement of gcc with aag at 1729-1731)"
     CDS             1111..1629
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     protein_bind    1750..1874
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1911..1941
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1995..2651
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             2996..3298
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3342..3466)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3492..3744
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3753..3769)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3982..4437
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4719..4823
                     /label=AmpR promoter
     CDS             4824..5681
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5855..6443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 catgcctgca ggtccccaga ttagcctttt caatttcaga aagaatgcta acccacagat
      301 ggttagagag gcttacgcag caggtctcat caagacgatc tacccgagca ataatctcca
      361 ggaaatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg
      421 catcaagaac acagagaaag atatatttct caagatcaga agtactattc cagtatggac
      481 gattcaaggc ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt
      541 agttcccact gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact
      601 cgccgtaaag actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa
      661 aatcttcgtc aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac
      721 agtctcagaa gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct
      781 cctcggattc cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg
      841 tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc
      901 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
      961 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
     1021 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
     1081 ggagagaaca cgggggactc tagaatgagc atggtgagca agggcgagga gctgttcacc
     1141 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1201 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     1261 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcctgcag
     1321 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1381 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1441 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1501 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1561 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1621 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1681 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1741 gctgttatca caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc
     1801 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
     1861 atccagtcac tatggcggcc cctcgagggg ccgcattagg caccccaggc tttacacttt
     1921 atgcttccgg ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag
     1981 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     2041 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
     2101 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     2161 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg
     2221 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     2281 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     2341 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     2401 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt
     2461 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     2521 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     2581 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     2641 agggcggggc gtaaacgcgt ggatccggct tactaaaagc cagataacag tatgcgtatt
     2701 tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa
     2761 aaagaggtat gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt
     2821 gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa
     2881 gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc
     2941 gcccggttta ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca
     3001 gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag
     3061 tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct
     3121 gtcagataaa gtctcccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg
     3181 catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga
     3241 tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata
     3301 aatgtcaggc tcccttatac acagccagtc tgcaggtcga ccatagtgac tggatatgtt
     3361 gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat
     3421 ttatatcatt ttacgtttct cgttcagctt tcttgtacaa agtggtgata acctagagct
     3481 cgaatttccc cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg
     3541 ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta
     3601 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat
     3661 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg
     3721 cggtgtcatc tatgttacta gatcgggaat tcactggccg tcgttttaca acgtcgtgac
     3781 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     3841 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     3901 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3961 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4021 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4081 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4141 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4201 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4261 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4321 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4381 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     4441 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4501 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4561 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4621 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4681 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4741 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4801 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4861 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4921 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4981 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5041 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5101 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5161 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5221 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5281 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5341 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5401 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5461 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5521 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5581 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5641 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5701 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5761 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5821 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5881 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5941 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6001 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6061 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6121 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6181 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6241 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6301 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6361 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6421 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6481 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6541 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6601 agtcagtgag cgaggaagcg gaag
//
