LOCUS       40924_33073        6516 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pnCGW DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6516)
  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 6516)
  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 6516)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6516)
  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..6516
                     /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             1111..1629
                     /codon_start=1
                     /label=VN173
                     /note="N-terminal fragment of mVenus for use in bimolecular
                     fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIE"
     protein_bind    1642..1766
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1803..1833
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1887..2543
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2888..3190
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3234..3358)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3384..3636
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3645..3661)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3874..4329
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4611..4715
                     /label=AmpR promoter
     CDS             4716..5573
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5747..6335
                     /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 ccctgaagtt catctgcacc
     1261 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag
     1321 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1381 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1441 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1501 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacat cagccacaac
     1561 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1621 aacatcgagg acgctgttat cacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat
     1681 gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt
     1741 aaaacacaac atatccagtc actatggcgg cccctcgagg ggccgcatta ggcaccccag
     1801 gctttacact ttatgcttcc ggctcgtata atgtgtggat tttgagttag gatccgtcga
     1861 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     1921 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     1981 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     2041 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     2101 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     2161 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     2221 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     2281 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     2341 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     2401 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     2461 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     2521 gcgatgagtg gcagggcggg gcgtaaacgc gtggatccgg cttactaaaa gccagataac
     2581 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
     2641 gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga
     2701 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
     2761 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
     2821 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc
     2881 tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     2941 ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag
     3001 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
     3061 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
     3121 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     3181 ctggggaata taaatgtcag gctcccttat acacagccag tctgcaggtc gaccatagtg
     3241 actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
     3301 atatattgat atttatatca ttttacgttt ctcgttcagc tttcttgtac aaagtggtga
     3361 taacctagag ctcgaatttc cccgatcgtt caaacatttg gcaataaagt ttcttaagat
     3421 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
     3481 atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
     3541 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
     3601 aattatcgcg cgcggtgtca tctatgttac tagatcggga attcactggc cgtcgtttta
     3661 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3721 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3781 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3841 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     3901 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3961 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4021 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4081 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4141 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4201 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4261 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     4321 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     4381 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4441 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4501 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     4561 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4621 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4681 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4741 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4801 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4861 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4921 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4981 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5041 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5101 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5161 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5221 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5281 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5341 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5401 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5461 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5521 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5581 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5641 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5701 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5761 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5821 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5881 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5941 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6001 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6061 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6121 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6181 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6241 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6301 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6361 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6421 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6481 ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
