LOCUS       40924_33497        6489 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pnRGW DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6489)
  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 6489)
  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 6489)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6489)
  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..6489
                     /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..1605
                     /label=nmrfp1
                     /note="nmrfp1"
     CDS             1114..1143
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    1615..1739
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1776..1806
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1860..2516
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2861..3163
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3207..3331)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3357..3609
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3618..3634)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3847..4302
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4584..4688
                     /label=AmpR promoter
     CDS             4689..5546
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5720..6308
                     /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 atggaacaaa agctaatctc cgaggaagac
     1141 ttgatggcct cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag
     1201 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     1261 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
     1321 atcctgtccc ctcagttcac gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     1381 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     1441 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     1501 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     1561 accatgggct gggaggcctc caccgagcgg atgtaccccg aggacgctgt tatcacaagt
     1621 ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
     1681 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
     1741 cggcccctcg aggggccgca ttaggcaccc caggctttac actttatgct tccggctcgt
     1801 ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag gaagctaaaa
     1861 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     1921 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     1981 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     2041 acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg
     2101 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     2161 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     2221 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     2281 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     2341 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     2401 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     2461 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa
     2521 cgcgtggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt
     2581 gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat
     2641 gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca aggcatatat
     2701 gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt
     2761 gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa
     2821 atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac
     2881 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac
     2941 gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc
     3001 ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga
     3061 tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga
     3121 aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct
     3181 tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt
     3241 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     3301 tttctcgttc agctttcttg tacaaagtgg tgataaccta gagctcgaat ttccccgatc
     3361 gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga
     3421 ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga
     3481 cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga
     3541 tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt
     3601 tactagatcg ggaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3661 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     3721 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     3781 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3841 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3901 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3961 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4021 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     4081 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4141 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     4201 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4261 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4321 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4381 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4441 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4501 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4561 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4621 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4681 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4741 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4801 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4861 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4921 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4981 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5041 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5101 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5161 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5221 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5281 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5341 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5401 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5461 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5521 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5581 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5641 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5701 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5761 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5821 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5881 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5941 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6001 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6061 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6121 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6181 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6241 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6301 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6361 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6421 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6481 aagcggaag
//
