LOCUS       .                       7987 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     CDS             1935..1937
                     /codon_start=1
                     /product="start codon for in vitro
                     transcription/translation"
                     /transl_table=1
                     /translation="M"
                     /label="ATG"
                     /note="color: #993366"
     misc_feature    1968..2035
                     /label="MCS"
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     rep_origin      6820..7984
                     /note="yeast 2μ plasmid origin of replication"
                     /note="color: #ffff00"
                     /label="2μ ori"
     CDS             complement(2719..3813)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /note="yeast auxotrophic marker"
                     /note="color: #ff7f50"
                     /product="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /transl_table=1
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIEATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA*"
                     /label="LEU2"
     CDS             complement(5573..6433)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #ccffcc"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        771..1475
                     /gene="S. cerevisiae ADH1"
                     /note="promoter for alcohol dehydrogenase 1"
                     /note="color: #ffffff"
                     /label="ADH1 promoter"
     rep_origin      complement(4814..5402)
                     /direction=LEFT
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="color: #ffff00"
                     /label="ori"
     promoter        complement(3814..4219)
                     /gene="S. cerevisiae LEU2"
                     /label="LEU2 promoter"
                     /note="color: #ffffff"
     CDS             1557..1898
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional
                     activator"
                     /transl_table=1
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
                     /label="GAL4 activation domain"
                     /note="color: #993366"
     terminator      2415..2602
                     /gene="S. cerevisiae ADH1"
                     /note="transcription terminator for alcohol dehydrogenase
                     1"
                     /note="color: #ffffff"
                     /label="ADH1 terminator"
     promoter        complement(6434..6538)
                     /gene="bla"
                     /label="AmpR promoter"
                     /note="color: #ffffff"
     CDS             1941..1967
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
                     /note="color: #cc99b2"
     promoter        1904..1922
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff"
                     /label="T7 promoter"
     CDS             1521..1541
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /transl_table=1
                     /translation="PKKKRKV"
                     /label="SV40 NLS"
                     /note="color: #993366"
     CDS             1491..1493
                     /codon_start=1
                     /product="start codon for expression in yeast"
                     /transl_table=1
                     /translation="M"
                     /label="ATG"
                     /note="color: #993366"
ORIGIN
        1 tgcatgcctg caggtcgaga tccgggatcg aagaaatgat ggtaaatgaa ataggaaatc
       61 aaggagcatg aaggcaaaag acaaatataa gggtcgaacg aaaaataaag tgaaaagtgt
      121 tgatatgatg tatttggctt tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca
      181 tgctgactct gtggcggacc cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc
      241 tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg
      301 agattggaga agcaataaga atgccggttg gggttgcgat gatgacgacc acgacaactg
      361 gtgtcattat ttaagttgcc gaaagaacct gagtgcattt gcaacatgag tatactagaa
      421 gaatgagcca agacttgcga gacgcgagtt tgccggtggt gcgaacaata gagcgaccat
      481 gaccttgaag gtgagacgcg cataaccgct agagtacttt gaagaggaaa cagcaatagg
      541 gttgctacca gtataaatag acaggtacat acaacactgg aaatggttgt ctgtttgagt
      601 acgctttcaa ttcatttggg tgtgcacttt attatgttac aatatggaag ggaactttac
      661 acttctccta tgcacatata ttaattaaag tccaatgcta gtagagaagg ggggtaacac
      721 ccctccgcgc tcttttccga tttttttcta aaccgtggaa tatttcggat atccttttgt
      781 tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga
      841 ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat
      901 agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc
      961 attgatggtg gtacataacg aactaatact gtagccctag acttgatagc catcatcata
     1021 tcgaagtttc actacccttt ttccatttgc catctattga agtaataata ggcgcatgca
     1081 acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat
     1141 gacaaaaaaa tgatggaaga cactaaagga aaaaattaac gacaaagaca gcaccaacag
     1201 atgtcgttgt tccagagctg atgaggggta tctcgaagca cacgaaactt tttccttcct
     1261 tcattcacgc acactactct ctaatgagca acggtatacg gccttccttc cagttacttg
     1321 aatttgaaat aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac ctgcaattat
     1381 taatcttttg tttcctcgtc attgttctcg ttccctttct tccttgtttc tttttctgca
     1441 caatatttca agctatacca agcatacaat caactccaag ctttgcaaag atggataaag
     1501 cggaattaat tcccgagcct ccaaaaaaga agagaaaggt cgaattgggt accgccgcca
     1561 attttaatca aagtgggaat attgctgata gctcattgtc cttcactttc actaacagta
     1621 gcaacggtcc gaacctcata acaactcaaa caaattctca agcgctttca caaccaattg
     1681 cctcctctaa cgttcatgat aacttcatga ataatgaaat cacggctagt aaaattgatg
     1741 atggtaataa ttcaaaacca ctgtcacctg gttggacgga ccaaactgcg tataacgcgt
     1801 ttggaatcac tacagggatg tttaatacca ctacaatgga tgatgtatat aactatctat
     1861 tcgatgatga agatacccca ccaaacccaa aaaaagagat ctttaatacg actcactata
     1921 gggcgagcgc cgccatggag tacccatacg acgtaccaga ttacgctcat atggccatgg
     1981 aggccagtga attccacccg ggtgggcatc gatacgggat ccatcgagct cgagctgcag
     2041 atgaatcgta gatactgaaa aaccccgcaa gttcacttca actgtgcatc gtgcaccatc
     2101 tcaatttctt tcatttatac atcgttttgc cttcttttat gtaactatac tcctctaagt
     2161 ttcaatcttg gccatgtaac ctctgatcta tagaattttt taaatgacta gaattaatgc
     2221 ccatcttttt tttggaccta aattcttcat gaaaatatat tacgagggct tattcagaag
     2281 ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta
     2341 ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg
     2401 acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     2461 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     2521 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     2581 ttattgacca cacctctacc ggccggtcga aattccccta ccctatgaac atattccatt
     2641 ttgtaatttc gtgtcgtttc tattatgaat ttcatttata aagtttatgt acaaatatca
     2701 taaaaaaaga gaatcttttt aagcaaggat tttcttaact tcttcggcga cagcatcacc
     2761 gacttcggtg gtactgttgg aaccacctaa atcaccagtt ctgatacctg catccaaaac
     2821 ctttttaact gcatcttcaa tggccttacc ttcttcaggc aagttcaatg acaatttcaa
     2881 catcattgca gcagacaaga tagtggcgat agggttgacc ttattctttg gcaaatctgg
     2941 agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc
     3001 caaggacgca gatggcaaca aacccaagga acctgggata acggaggctt catcggagat
     3061 gatatcacca aacatgttgc tggtgattat aataccattt aggtgggttg ggttcttaac
     3121 taggatcatg gcggcagaat caatcaattg atgttgaacc ttcaatgtag gaaattcgtt
     3181 cttgatggtt tcctccacag tttttctcca taatcttgaa gaggccaaaa cattagcttt
     3241 atccaaggac caaataggca atggtggctc atgttgtagg gccatgaaag cggccattct
     3301 tgtgattctt tgcacttctg gaacggtgta ttgttcacta tcccaagcga caccatcacc
     3361 atcgtcttcc tttctcttac caaagtaaat acctcccact aattctctga caacaacgaa
     3421 gtcagtacct ttagcaaatt gtggcttgat tggagataag tctaaaagag agtcggatgc
     3481 aaagttacat ggtcttaagt tggcgtacaa ttgaagttct ttacggattt ttagtaaacc
     3541 ttgttcaggt ctaacactac ctgtacccca tttaggacca cccacagcac ctaacaaaac
     3601 ggcatcagcc ttcttggagg cttccagcgc ctcatctgga agtgggacac ctgtagcttc
     3661 gatagcagca ccaccaatta aatgattttc gaaatcgaac ttgacattgg aacgaacatc
     3721 agaaatagct ttaagaacct taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc
     3781 tggcaaaacg acgatcttct taggggcaga cattagaatg gtatatcctt gaaatatata
     3841 tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat tgctaaccac
     3901 ctattggaaa aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa
     3961 gcatttagtc atgaacgctt ctctattcta tatgaaaagc cggttccggc gctctcacct
     4021 ttcctttttc tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat
     4081 taacaaaaaa tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc
     4141 gttatgttga ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata
     4201 cctgagtatt cccacagttg ggggatctcg actctagcta gaggatcaat tcgtaatcat
     4261 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4321 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4381 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctga taacttcgta
     4441 taatgtatgc tatacgaagt tattaggtct gaagaggagt ttacgtccag ccaagctagc
     4501 ttggctgcag gtcgagcggc cgcgatccgg aacccttaat ataacttcgt ataatgtatg
     4561 ctatacgaag ttatcagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4621 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4681 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4981 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5221 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5341 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5401 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5461 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5521 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5581 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5641 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5701 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5761 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5821 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5881 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5941 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6001 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6061 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6121 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6181 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6241 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6301 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6361 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6421 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6481 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6541 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     6601 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     6661 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     6721 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6781 atgcggcatc agagcagatt gtactgagag tgcaccataa cgcatttaag cataaacacg
     6841 cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac
     6901 gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg
     6961 aaacgctttg aagttcctat tccgaagttc ctattctcta gctagaaagt ataggaactt
     7021 cagagcgctt ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc
     7081 accggactgt aacgagctac taaaatattg cgaataccgc ttccacaaac attgctcaaa
     7141 agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc atccaccttt
     7201 cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat ctctagtatt
     7261 actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga aaacaatacg
     7321 aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa ccgttcataa
     7381 ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag tatgcgcaat
     7441 ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc acccgcagct
     7501 tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga agagaaaata
     7561 gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag
     7621 actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa
     7681 tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct ttgttggtaa
     7741 aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
     7801 aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag attcttcgtt
     7861 ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg
     7921 tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc acagatgctt
     7981 cgttgct
//
