LOCUS       40924_23974        6640 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pGWnV4 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  AUTHORS   Nishimura K, Matsunami E, Yoshida S, Kohata S, Yamauchi J, Jisaka M,
            Nagaya T, Yokota K, Nakagawa T.
  TITLE     The tyrosine-sorting motif of the vacuolar sorting receptor VSR4 
            from Arabidopsis thaliana, which is involved in the interaction 
            between VSR4 and AP1M2, mu1-adaptin type 2 of clathrin adaptor 
            complex 1 subunits, participates in the post-Golgi sorting of VSR4
  JOURNAL   Biosci. Biotechnol. Biochem. 80 (4), 694-705 (2016)
  PUBMED    26745465
REFERENCE   2  (bases 1 to 6640)
  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 6640)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6640)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biosci. 
            Biotechnol. Biochem."; date: "2016"; volume: "80"; issue: "4"; 
            pages: "694-705"
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..6640
                     /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        763..1108
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    1130..1254
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1291..1321
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1375..2031
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             2376..2678
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2722..2846)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2857..3486
                     /note="n-terminal fragment of yellow fluorescent protein 
                     (replacement of gcc with aag at 3475-3477)"
     CDS             2857..3375
                     /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"
     terminator      3508..3760
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3769..3785)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3998..4453
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4735..4839
                     /label=AmpR promoter
     CDS             4840..5697
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5871..6459
                     /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 ggtctagacc caagcttgca tgcctgcagg tccccagatt agccttttca atttcagaaa
      301 gaatgctaac ccacagatgg ttagagaggc ttacgcagca ggtctcatca agacgatcta
      361 cccgagcaat aatctccagg aaatcaaata ccttcccaag aaggttaaag atgcagtcaa
      421 aagattcagg actaactgca tcaagaacac agagaaagat atatttctca agatcagaag
      481 tactattcca gtatggacga ttcaaggctt gcttcacaaa ccaaggcaag taatagagat
      541 tggagtctct aaaaaggtag ttcccactga atcaaaggcc atggagtcaa agattcaaat
      601 agaggaccta acagaactcg ccgtaaagac tggcgaacag ttcatacaga gtctcttacg
      661 actcaatgac aagaagaaaa tcttcgtcaa catggtggag cacgacacac ttgtctactc
      721 caaaaatatc aaagatacag tctcagaaga ccaaagggca attgagactt ttcaacaaag
      781 ggtaatatcc ggaaacctcc tcggattcca ttgcccagct atctgtcact ttattgtgaa
      841 gatagtggaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggccat
      901 cgttgaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
      961 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc
     1021 cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata
     1081 aggaagttca tttcatttgg agagaacacg ggggactcta gagttatcaa caagtttgta
     1141 caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt
     1201 gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac tatggcggcc
     1261 cctcgagggg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
     1321 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
     1381 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     1441 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     1501 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     1561 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
     1621 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     1681 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     1741 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     1801 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1861 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     1921 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     1981 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaagatct
     2041 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     2101 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc
     2161 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     2221 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     2281 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     2341 cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata
     2401 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     2461 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     2521 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     2581 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     2641 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
     2701 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta
     2761 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     2821 cgttcagctt tcttgtacaa agtggctgat aacagcatgg tgagcaaggg cgaggagctg
     2881 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2941 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
     3001 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
     3061 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3121 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     3181 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3241 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3301 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     3361 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     3421 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     3481 agcaaagctt agagctcgaa tttccccgat cgttcaaaca tttggcaata aagtttctta
     3541 agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt
     3601 aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt
     3661 agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag
     3721 gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattcac tggccgtcgt
     3781 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     3841 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     3901 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     3961 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     4021 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4081 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4141 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4201 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4261 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4321 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     4381 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4441 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4501 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4561 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4621 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4681 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4741 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4801 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4861 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4921 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4981 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5041 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5101 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5161 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5221 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5281 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5341 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5401 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5461 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5521 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5581 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5641 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5701 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5761 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5821 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5881 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5941 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6001 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     6061 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6121 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6181 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6241 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6301 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6361 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6421 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6481 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6541 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6601 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
