LOCUS       62056_8410        8979 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8979)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    15..31
                     /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     39..55
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        76..94
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        863..1567
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             2555..2788
                     /codon_start=1
                     /label=VP16 AD
                     /note="transcriptional activation domain of herpes simplex
                     virus protein VP16 (Triezenberg et al., 1988; Cousens et 
                     al., 1989)"
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     terminator      2793..3037
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(3060..3078)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3088..3104)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3249..3704
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4004..4408
                     /label=LEU2 promoter
     CDS             4421..5512
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     CDS             complement(6236..7048)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      7490..8078
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(8255..8756)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     protein_bind    8920..8941
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8956..8979
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
       61 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tcgccgggat
      121 cgaagaaatg atggtaaatg aaataggaaa tcaaggagca tgaaggcaaa agacaaatat
      181 aagggtcgaa cgaaaaataa agtgaaaagt gttgatatga tgtatttggc tttgcggcgc
      241 cgaaaaaacg agtttacgca attgcacaat catgctgact ctgtggcgga cccgcgctct
      301 tgccggcccg gcgataacgc tgggcgtgag gctgtgcccg gcggagtttt ttgcgcctgc
      361 attttccaag gtttaccctg cgctaagggg cgagattgga gaagcaataa gaatgccggt
      421 tggggttgcg atgatgacga ccacgacaac tggtgtcatt atttaagttg ccgaaagaac
      481 ctgagtgcat ttgcaacatg agtatactag aagaatgagc caagacttgc gagacgcgag
      541 tttgccggtg gtgcgaacaa tagagcgacc atgaccttga aggtgagacg cgcataaccg
      601 ctagagtact ttgaagagga aacagcaata gggttgctac cagtataaat agacaggtac
      661 atacaacact ggaaatggtt gtctgtttga gtacgctttc aattcatttg ggtgtgcact
      721 ttattatgtt acaatatgga agggaacttt acacttctcc tatgcacata tattaattaa
      781 agtccaatgc tagtagagaa ggggggtaac acccctccgc gctcttttcc gatttttttc
      841 taaaccgtgg aatatttcgg atatcctttt gttgtttccg ggtgtacaat atggacttcc
      901 tcttttctgg caaccaaacc catacatcgg gattcctata ataccttcgt tggtctccct
      961 aacatgtagg tggcggaggg gagatataca atagaacaga taccagacaa gacataatgg
     1021 gctaaacaag actacaccaa ttacactgcc tcattgatgg tggtacataa cgaactaata
     1081 ctgtagccct agacttgata gccatcatca tatcgaagtt tcactaccct ttttccattt
     1141 gccatctatt gaagtaataa taggcgcatg caacttcttt tctttttttt tcttttctct
     1201 ctcccccgtt gttgtctcac catatccgca atgacaaaaa aatgatggaa gacactaaag
     1261 gaaaaaatta acgacaaaga cagcaccaac agatgtcgtt gttccagagc tgatgagggg
     1321 tatctcgaag cacacgaaac tttttccttc cttcattcac gcacactact ctctaatgag
     1381 caacggtata cggccttcct tccagttact tgaatttgaa ataaaaaaaa gtttgctgtc
     1441 ttgctatcaa gtataaatag acctgcaatt attaatcttt tgtttcctcg tcattgttct
     1501 cgttcccttt cttccttgtt tctttttctg cacaatattt caagctatac caagcataca
     1561 atcaactcca agctggccgc tctagagcaa gcaaaagatg atctctgatg aacagctgaa
     1621 ctccttggcc atcaccttcg gtattgtgat gatgacttta attgtcattt accatgctgt
     1681 tgactccacc atgtctccta agaacgcggc cattacggcc aggcctccat ggtatatctt
     1741 ggccgcctcg gccatgtcgg gggggatccc tccagatcaa caaagattga tctttgccgg
     1801 taagcagcta gaagacggta gaacgctgtc tgattacaac attcagaagg agtccacctt
     1861 acatcttgtg ctaaggctaa gaggtggtat gcacagatca gcttgcggcc gcatgaaagc
     1921 gttaacggcc aggcaacaag aggtgtttga tctagaagga gtccacctta ccatccgtga
     1981 tcacatcagc cagacaggta tgccgccgac gcgtgcggaa atcgcgcagc gtttggggtt
     2041 ccgttcccca aacgcggctg aagaacatct gaaggcgctg gcacgcaaag gcgttattga
     2101 aattgtttcc ggcgcatcac gcgggatccg tctgttgcag gaagaggaag aagggttgcc
     2161 gctggtaggt cgtgtggctg ccggtgaacc acttctggcg caacagcata ttgaaggtca
     2221 ttatcaggtc gacccttcct tattcaagcc gaatgctgat ttcctgctgc gcgtcagcgg
     2281 gatgtcgatg aaagatatcg gcattatgga tggtgacttg ctggcagtgc ataaaactca
     2341 ggatgtacgt aacggtcagg tcgttgtcgc acgtattgat gacgaagtta ccgttaaggg
     2401 cctgaaaaaa cagggcaata aagtcgaact gttgccagaa aatagcgagt ttaaaccaat
     2461 tgtcgttgac cttcgtcagc agagcttcac cattgaaggg ctggcggttg gggttattcg
     2521 caacggcgac tggctggaat tcccggggat atgggccccc ccgaccgatg tcagcctggg
     2581 ggacgagctc cacttagacg gcgaggacgt ggcgatggcg catgccgacg cgctagacga
     2641 tttcgatctg gacatgttgg gggacgggga ttccccgggt ccgggattta ccccccacga
     2701 ctccgccccc tacggcgctc tggatatggc cgacttcgag tttgagcaga tgtttaccga
     2761 tgcccttgga attgacgagt acggtgggta ggtcatgtaa ttagttatgt cacgcttaca
     2821 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     2881 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     2941 aatttttctt tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg
     3001 cttgagaagg ttttgggacg ctcgaaggtt taatttgggg ggcccggtac ccaattcgcc
     3061 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     3121 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3181 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3241 atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3301 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3361 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3421 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3481 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3541 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3601 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3661 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcctgat gcggtatttt
     3721 ctccttacgc atctgtgcgg tatttcacac cgcatatcga cggtcgagga gaacttctag
     3781 tatatccaca tacctaatat tattgcctta ttaaaaatgg aatcccaaca attacatcaa
     3841 aatccacatt ctcttcaaaa tcaattgtcc tgtacttcct tgttcatgtg tgttcaaaaa
     3901 cgttatattt ataggataat tatactctat ttctcaacaa gtaattggtt gtttggccga
     3961 gcggtctaag gcgcctgatt caagaaatat cttgaccgca gttaactgtg ggaatactca
     4021 ggtatcgtaa gatgcaagag ttcgaatctc ttagcaacca ttattttttt cctcaacata
     4081 acgagaacac acaggggcgc tatcgcacag aatcaaattc gatgactgga aattttttgt
     4141 taatttcaga ggtcgcctga cgcatatacc tttttcaact gaaaaattgg gagaaaaagg
     4201 aaaggtgaga ggccggaacc ggcttttcat atagaataga gaagcgttca tgactaaatg
     4261 cttgcatcac aatacttgaa gttgacaata ttatttaagg acctattgtt ttttccaata
     4321 ggtggttagc aatcgtctta ctttctaact tttcttacct tttacatttc agcaatatat
     4381 atatatattt caaggatata ccattctaat gtctgcccct atgtctgccc ctaagaagat
     4441 cgtcgttttg ccaggtgacc acgttggtca agaaatcaca gccgaagcca ttaaggttct
     4501 taaagctatt tctgatgttc gttccaatgt caagttcgat ttcgaaaatc atttaattgg
     4561 tggtgctgct atcgatgcta caggtgtccc acttccagat gaggcgctgg aagcctccaa
     4621 gaaggttgat gccgttttgt taggtgctgt gggtggtcct aaatggggta ccggtagtgt
     4681 tagacctgaa caaggtttac taaaaatccg taaagaactt caattgtacg ccaacttaag
     4741 accatgtaac tttgcatccg actctctttt agacttatct ccaatcaagc cacaatttgc
     4801 taaaggtact gacttcgttg ttgtcagaga attagtggga ggtatttact ttggtaagag
     4861 aaaggaagac gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt
     4921 gcaaagaatc acaagaatgg ccgctttcat ggccctacaa catgagccac cattgcctat
     4981 ttggtccttg gataaagcta atgttttggc ctcttcaaga ttatggagaa aaactgtgga
     5041 ggaaaccatc aagaacgaat tccctacatt gaaggttcaa catcaattga ttgattctgc
     5101 cgccatgatc ctagttaaga acccaaccca cctaaatggt attataatca ccagcaacat
     5161 gtttggtgat atcatctccg atgaagcctc cgttatccca ggttccttgg gtttgttgcc
     5221 atctgcgtcc ttggcctctt tgccagacaa gaacaccgca tttggtttgt acgaaccatg
     5281 ccacggttct gctccagatt tgccaaagaa taaggttgac cctatcgcca ctatcttgtc
     5341 tgctgcaatg atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg ccattgaaga
     5401 tgcagttaaa aaggttttgg atgcaggtat cagaactggt gatttaggtg gttccaacag
     5461 taccaccgaa gtcggtgatg ctgtcgccga agaagttaag aaaatccttg cttaaaaaga
     5521 ttctcttttt ttatgatatt tgtacataaa ctttataaat gaaattcata atagaaacga
     5581 cacgaaatta caaaatggaa tatgttcata gggtagacga aactatatac gcaatctaca
     5641 tacatttatc aagaaggaga aaaaggagga tagtaaagga atacaggtaa gcaaattgat
     5701 actaatggct caacgtgata aggaaaaaga attgcacttt aacattaata ttgacaagga
     5761 ggagggcacc acacaaaaag ttaggtgtaa cagaaaatca tgaaactacg attcctaatt
     5821 tgatattgga ggattttctc taaaaaaaaa aaaatacaac aaataaaaaa cactcaatga
     5881 cctgaccatt tgatggagtt taagtcaata ccttcttgaa ccatttccca taatggtgaa
     5941 agttccctca agaattttac tctgtcagaa acggccttac gacgtagtcg atatggtgca
     6001 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     6061 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     6121 ccgtctccgg gaccacatgt ggatgatcca gttcgattta ttcaacaaag ccgccgtccc
     6181 gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa
     6241 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     6301 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     6361 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     6421 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     6481 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     6541 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     6601 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     6661 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     6721 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     6781 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     6841 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     6901 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     6961 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgagc
     7021 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     7081 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     7141 gagacacaac gtggcttact aggatcgatc cgacatgtgg cggtatcatt gcagcactgg
     7201 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     7261 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     7321 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7381 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7441 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7501 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7561 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7621 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7681 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7741 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7801 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7861 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7921 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7981 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     8041 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     8101 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     8161 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     8221 cgaccgagcg cagcgagtca gtgagcgagg aagctcgctt gcctgtaact tacacgcgcc
     8281 tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt
     8341 tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa
     8401 aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt
     8461 agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat
     8521 cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac
     8581 ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt
     8641 tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt
     8701 aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa
     8761 aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctaagcgg aagagcgccc
     8821 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     8881 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca
     8941 ttaggcaccc caggctttac actttatgct tccggctcc
//
