LOCUS       40924_33612        8001 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast two-hybrid vector pNtrp DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8001)
  AUTHORS   Banno H.
  TITLE     Identification of proteins that interact with a plant nuclear 
            protein using the yeast split-Trp sensor
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8001)
  AUTHORS   Banno H.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-APR-2014) Contact:Hiroharu Banno Chubu University, 
            Bioscience and Biotechnology; 1200 Matsumoto-cho, Kasugai, Aichi 
            487-8501, Japan Phone :81-568-52-6242
REFERENCE   3  (bases 1 to 8001)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8001)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-APR-2014) Contact:Hiroharu Banno Chubu University, Bioscience 
            and Biotechnology"; volume: " 1200 Matsumoto-cho, Kasugai, Aichi 
            487-8501, Japan Phone "; pages: "81-568-52-624"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..1165)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1447..1551
                     /label=AmpR promoter
     CDS             1552..2409
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2583..3171
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3479..3907
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     protein_bind    3969..3990
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4005..4035
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4043..4059
                     /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     4067..4083
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4125..4529
                     /label=LEU2 promoter
     CDS             4530..5621
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        6496..7200
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_feature    7209..7340
                     /label=Ntrp
                     /note="Ntrp"
     CDS             7341..7367
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     misc_feature    7371..7423
                     /label=multiple cloning site(MCS)
                     /note="multiple cloning site(MCS)"
     primer_bind     complement(7447..7463)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      7670..7857
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
ORIGIN
        1 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
       61 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
      121 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
      181 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
      241 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
      301 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
      361 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
      421 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
      481 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
      541 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
      601 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
      661 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
      721 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
      781 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
      841 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
      901 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc
      961 tctgaagttc ctatactttc tagctagaga ataggaactt cggaatagga acttcaaagc
     1021 gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac agctcactgt
     1081 tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga agaacggcat
     1141 agtgcgtgtt tatgcttaaa tgcgttatgg tgcactctca gtacaatctg ctctgatgcc
     1201 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     1261 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     1321 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     1381 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     1441 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1501 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1561 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1621 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1681 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1741 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     1801 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1861 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     1921 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     1981 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2041 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2101 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2161 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2221 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2281 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2341 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2401 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2461 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2521 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2581 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2641 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2701 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     2761 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2821 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2881 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2941 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3001 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3061 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3121 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3181 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3241 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3301 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     3361 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatccagctg gcgtaatagc
     3421 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     3481 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3541 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3601 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3661 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3721 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3781 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3841 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3901 gcgaatttta acaaaatctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     3961 aacgcaatta atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt
     4021 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     4081 gaccatgatt acgaattgat cctctagcta gagtcgagat ccccaactgt gggaatactc
     4141 aggtatcgta agatgcaaga gttcgaatct cttagcaacc attatttttt tcctcaacat
     4201 aacgagaaca cacaggggcg ctatcgcaca gaatcaaatt cgatgactgg aaattttttg
     4261 ttaatttcag aggtcgcctg acgcatatac ctttttcaac tgaaaaattg ggagaaaaag
     4321 gaaaggtgag aggccggaac cggcttttca tatagaatag agaagcgttc atgactaaat
     4381 gcttgcatca caatacttga agttgacaat attatttaag gacctattgt tttttccaat
     4441 aggtggttag caatcgtctt actttctaac ttttcttacc ttttacattt cagcaatata
     4501 tatatatatt tcaaggatat accattctaa tgtctgcccc taagaagatc gtcgttttgc
     4561 caggtgacca cgttggtcaa gaaatcacag ccgaagccat taaggttctt aaagctattt
     4621 ctgatgttcg ttccaatgtc aagttcgatt tcgaaaatca tttaattggt ggtgctgcta
     4681 tcgatgctac aggtgtccca cttccagatg aggcgctgga agcctccaag aaggttgatg
     4741 ccgttttgtt aggtgctgtg ggtggtccta aatggggtac aggtagtgtt agacctgaac
     4801 aaggtttact aaaaatccgt aaagaacttc aattgtacgc caacttaaga ccatgtaact
     4861 ttgcatccga ctctctttta gacttatctc caatcaagcc acaatttgct aaaggtactg
     4921 acttcgttgt tgtcagagaa ttagtgggag gtatttactt tggtaagaga aaggaagacg
     4981 atggtgatgg tgtcgcttgg gatagtgaac aatacaccgt tccagaagtg caaagaatca
     5041 caagaatggc cgctttcatg gccctacaac atgagccacc attgcctatt tggtccttgg
     5101 ataaagctaa tgttttggcc tcttcaagat tatggagaaa aactgtggag gaaaccatca
     5161 agaacgaatt tcctacattg aaggttcaac atcaattgat tgattctgcc gccatgatcc
     5221 tagttaagaa cccaacccac ctaaatggta ttataatcac cagcaacatg tttggtgata
     5281 tcatctccga tgaagcctcc gttatcccag gttccttggg tttgttgcca tctgcgtcct
     5341 tggcctcttt gccagacaag aacaccgcat ttggtttgta cgaaccatgc cacggttctg
     5401 ctccagattt gccaaagaat aaggttgacc ctatcgccac tatcttgtct gctgcaatga
     5461 tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc cattgaagat gcagttaaaa
     5521 aggttttgga tgcaggtatc agaactggtg atttaggtgg ttccaacagt accaccgaag
     5581 tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc ttaaaaagat tctctttttt
     5641 tatgatattt gtacataaac tttataaatg aaattcataa tagaaacgac acgaaattac
     5701 aaaatggaat atgttcatag ggtaggggaa tttcgaccgg atcgatccgg gatcgaagaa
     5761 atgatggtaa atgaaatagg aaatcaagga gcatgaaggc aaaagacaaa tataagggtc
     5821 gaacgaaaaa taaagtgaaa agtgttgata tgatgtattt ggctttgcgg cgccgaaaaa
     5881 acgagtttac gcaattgcac aatcatgctg actctgtggc ggacccgcgc tcttgccggc
     5941 ccggcgataa cgctgggcgt gaggctgtgc ccggcggagt tttttgcgcc tgcattttcc
     6001 aaggtttacc ctgcgctaag gggcgagatt ggagaagcaa taagaatgcc ggttggggtt
     6061 gcgatgatga cgaccacgac aactggtgtc attatttaag ttgccgaaag aacctgagtg
     6121 catttgcaac atgagtatac tagaagaatg agccaagact tgcgagacgc gagtttgccg
     6181 gtggtgcgaa caatagagcg accatgacct tgaaggtgag acgcgcataa ccgctagagt
     6241 actttgaaga ggaaacagca atagggttgc taccagtata aatagacagg tacatacaac
     6301 actggaaatg gttgtctgtt tgagtacgct ttcaattcat ttgggtgtgc actttattat
     6361 gttacaatat ggaagggaac tttacacttc tcctatgcac atatattaat taaagtccaa
     6421 tgctagtaga gaaggggggt aacacccctc cgcgctcttt tccgattttt ttctaaaccg
     6481 tggaatattt cggatatcct tttgttgttt ccgggtgtac aatatggact tcctcttttc
     6541 tggcaaccaa acccatacat cgggattcct ataatacctt cgttggtctc cctaacatgt
     6601 aggtggcgga ggggagatat acaatagaac agataccaga caagacataa tgggctaaac
     6661 aagactacac caattacact gcctcattga tggtggtaca taacgaacta atactgtagc
     6721 cctagacttg atagccatca tcatatcgaa gtttcactac cctttttcca tttgccatct
     6781 attgaagtaa taataggcgc atgcaacttc ttttcttttt ttttcttttc tctctccccc
     6841 gttgttgtct caccatatcc gcaatgacaa aaaaatgatg gaagacacta aaggaaaaaa
     6901 ttaacgacaa agacagcacc aacagatgtc gttgttccag agctgatgag gggtatctcg
     6961 aagcacacga aactttttcc ttccttcatt cacgcacact actctctaat gagcaacggt
     7021 atacggcctt ccttccagtt acttgaattt gaaataaaaa aaagtttgct gtcttgctat
     7081 caagtataaa tagacctgca attattaatc ttttgtttcc tcgtcattgt tctcgttccc
     7141 tttcttcctt gtttcttttt ctgcacaata tttcaagcta taccaagcat acaatcaact
     7201 ccaagcttat gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg
     7261 gcttgcagag cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta
     7321 ttatatgtgt gcccaataga tacccttatg atgtgccaga ttatgcctct gtcgacggcg
     7381 cgccggccac ttcggcctcg agagatctgg ccgaagcggc cgctagatag atagaagctc
     7441 gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     7501 taatcgcctt gcagcacatc cccctttcgc cagagctttg gacttcttcg ccagaggttt
     7561 ggtcaagtct ccaatcaagg ttgtcggctt gtctaccttg ccagaaattt acgaaaagat
     7621 ggaaaagggt caaatcgttg gtagatacgt tgttgacact tctaaataag cgaatttctt
     7681 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     7741 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     7801 caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat
     7861 gccgagcaaa tgcctgcaaa tcgctcccca tttcacccaa ttgtagatat gctaactcca
     7921 gcaatgagtt gatgaatctc ggtgtgtatt ttatgtcctc agaggacaac acctgttgta
     7981 atcgttcttc cacacggaag c
//
