LOCUS       40924_13400        6237 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Gateway vector pcRGW DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6237)
  AUTHORS   Kamigaki A, Nito K, Hikino K, Goto-Yamada S, Nishimura M, Nakagawa 
            T, Mano S.
  TITLE     Gateway Vectors for Simultaneous Detection of Multiple 
            Protein-Protein Interactions in Plant Cells Using Bimolecular 
            Fluorescence Complementation
  JOURNAL   PLoS ONE 11 (8), E0160717 (2016)
  PUBMED    27490375
REFERENCE   2  (bases 1 to 6237)
  AUTHORS   Mano S, Nakagawa T.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUL-2013) Contact:Shoji Mano National Institute for 
            Basic Biology, Department of Cell Biology; 38 Nishigonaka, Myodaiji,
            Okazaki, Aichi 444-8585, Japan
REFERENCE   3  (bases 1 to 6237)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6237)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2016"; volume: "11"; issue: "8"; pages: "E0160717"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-JUL-2013) Contact:Shoji Mano National Institute for Basic 
            Biology, Department of Cell Biology; 38 Nishigonaka, Myodaiji, 
            Okazaki, Aichi 444-8585, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     These colnes were obtained at our laboratory.
FEATURES             Location/Qualifiers
     source          1..6237
                     /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"
     CDS             1114..1140
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     protein_bind    1363..1487
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1524..1554
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1608..2264
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2609..2911
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2955..3079)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3105..3357
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3366..3382)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3595..4050
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4332..4436
                     /label=AmpR promoter
     CDS             4437..5294
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5468..6056
                     /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 atgtacccat acgatgttcc tgactatgcg
     1141 ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg ccactacgac
     1201 gccgaggtca agaccaccta catggccaag aagcccgtgc agctgcccgg cgcctacaag
     1261 accgacatca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1321 gagcgcgccg agggccgcca ctccaccggc gccgctgtta tcacaagttt gtacaaaaaa
     1381 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
     1441 aaacagacta cataatactg taaaacacaa catatccagt cactatggcg gcccctcgag
     1501 gggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
     1561 ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     1621 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     1681 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1741 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1801 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     1861 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1921 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1981 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     2041 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     2101 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     2161 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     2221 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg
     2281 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     2341 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
     2401 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     2461 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     2521 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     2581 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     2641 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     2701 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     2761 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     2821 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     2881 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
     2941 gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     3001 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     3061 ctttcttgta caaagtggtg ataacctaga gctcgaattt ccccgatcgt tcaaacattt
     3121 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
     3181 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
     3241 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
     3301 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcggg
     3361 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     3421 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     3481 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     3541 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3601 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3661 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3721 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3781 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3841 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3901 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     3961 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4021 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     4081 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4141 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4201 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     4261 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     4321 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4381 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4441 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4501 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4561 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4621 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4681 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4741 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4801 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4861 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4921 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4981 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5041 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5101 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5161 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5221 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5281 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5341 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5401 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5461 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5521 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5581 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     5641 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5701 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5761 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5821 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5881 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5941 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6001 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6061 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6121 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6181 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
