LOCUS       .                       7697 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     rep_origin      complement(237..825)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(1294..1672)
                     /label="ISS"
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of
                     5'-AACGTT-3' (Sato et al., 1996)"
     CDS             complement(1955..2755)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(2756..2976)
                     /label="URA3 promoter"
     promoter        2985..3328
                     /label="TEF promoter"
                     /note="Ashbya gossypii TEF promoter"
     CDS             3348..3911
                     /label="NrsR"
                     /note="nourseothricin acetyltransferase"
     terminator      3929..4126
                     /label="TEF terminator"
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(4723..5603)
                     /label="2μ ori"
                     /note="yeast 2μ plasmid origin of replication"
     rep_origin      complement(5614..6127)
                     /label="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6146..6588
                     /label="GAL1 promoter"
                     /note="inducible promoter, regulated by Gal4"
     promoter        6620..6638
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6670..7383
                     /label="GFP (S65T)"
                     /note="S65T variant of Aequorea victoria green fluorescent
                     protein (Heim et al., 1995)"
     terminator      7446..7693
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa gcccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1081 ataactacga tacgggagcg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1201 agaagtggtc ctgcaacttt atccgcctcc attcagtcta ttaattgttg ccgggaagct
     1261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttggcattgc tacaggcatc
     1321 gtggtgtcac tctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1621 aatagtgtat cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1861 ttcctttttc aatgggtaat aactgatata attaaattga agctctaatt tgtgagttta
     1921 gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat
     1981 atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg
     2041 caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg
     2101 ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa
     2161 tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa
     2221 caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata
     2281 gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt
     2341 cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg
     2401 taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat
     2461 taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg
     2521 cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt
     2581 ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac
     2641 gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg
     2701 cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt
     2761 atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact gggcaatttc
     2821 atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc
     2881 gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaagaaac cgaaatcaaa
     2941 aaaaagaata aaaaaaaaat gatgaattga attgaaaagc tagcgacatg gaggcccaga
     3001 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
     3061 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
     3121 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
     3181 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     3241 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     3301 ctcacatcac atccgaacat aaacaaccgg atcctacaat tatgtctact actttggatg
     3361 atactgctta tagatacaga acttctgttc caggtgatgc tgaagctatt gaagctttgg
     3421 atggttcttt cactaccgat actgttttta gagttactgc tactggtgat ggtttcactt
     3481 tgagagaagt tccagttgat ccaccattga ctaaggtttt cccagatgat gaatccgatg
     3541 atgaatccga tgctggtgaa gatggtgatc cagattctag aactttcgtt gcttatggtg
     3601 atgatggtga tttggctggt ttcgttgttg tttcttattc tggttggaac agaagattga
     3661 ctgttgaaga tattgaagtt gctccagaac atagaggtca tggtgttggt agagctttga
     3721 tgggtttggc tactgaattc gccagagaaa gaggtgctgg tcatttgtgg ttggaagtta
     3781 ccaatgttaa tgctccagct attcatgctt atagaagaat gggtttcact ttgtgtggtt
     3841 tggatactgc tttatacgat ggtactgctt ccgatggtga acaagctttg tatatgtcta
     3901 tgccatgtcc ataataactc gagtcagtac tgacaataaa aagattcttg ttttcaagaa
     3961 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     4021 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     4081 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactggcta gcttatcgat
     4141 gataagctgt caaagatgag aattaattcc acggactata gactatacta gatactccgt
     4201 ctactgtacg atacacttcc gctcaggtcc ttgtccttta acgaggcctt accactcttt
     4261 tgttactcta ttgatccagc tcagcaaagg cagtgtgatc taagattcta tcttcgcgat
     4321 gtagtaaaac tagctagacc gagaaagaga ctagaaatgc aaaaggcact tctacaatgg
     4381 ctgccatcat tattatccga tgtgacgctg cagcttctca atgatattcg aatacgcttt
     4441 gaggagatac agcctaatat ccgacaaact gttttacaga tttacgatcg tacttgttac
     4501 ccatcattga attttgaaca tccgaacctg ggagttttcc ctgaaacaga tagtatattt
     4561 gaacctgtat aataatatat agtctagcgc tttacggaag acaatgtatg tatttcggtt
     4621 cctggagaaa ctattgcatc tattgcatag gtaatcttgc acgtcgcatc cccggttcat
     4681 tttctgcgtt tccatcttgc acttcaatag catatctttg ttaacgaagc atctgtgctt
     4741 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     4801 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     4861 gcttcatttt tgtaaaacaa aaatgcaacg cgacgagagc gctaattttt caaacaaaga
     4921 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa
     4981 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca
     5041 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac
     5101 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt
     5161 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg
     5221 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac
     5281 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt
     5341 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt
     5401 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga
     5461 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg
     5521 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg
     5581 tttgtggaag cggtattcgc aatgggaagc tccaccccgg ttgataatca gaaaagcccc
     5641 aaaaacagga agattgtata agcaaatatt taaattgtaa acgttaatat tttgttaaaa
     5701 ttcgcgttaa atttttgtta aatcagctca ttttttaacg aatagcccga aatcggcaaa
     5761 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttccaac
     5821 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaag ggtctatcag
     5881 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5941 aaagcagtaa atcggaaggg taaacggatg cccccattta gagcttgacg gggaaagccg
     6001 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggggctag ggcggtggga
     6061 agtgtagggg tcacgctggg cgtaaccacc acacccgccg cgcttaatgg ggcgctacag
     6121 ggcgcgtggg gatgatccac tagtacggat tagaagccgc cgagcgggtg acagccctcc
     6181 gaaggaagac tctcctccgt gcgtcctcgt cctcaccggt cgcgttcctg aaacgcagat
     6241 gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct tttatggtta
     6301 tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaatgaac gaatcaaatt
     6361 aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg taattaatca
     6421 gcgaagcgat gatttttgat ctattaacag atatataaat gcaaaaactg cattaaccac
     6481 tttaactaat actttcaaca ttttcggttt gtattacttc ttattcaaat gtaataaaag
     6541 tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaaccccgg
     6601 atcggactac tagcagctgt aatacgactc actataggga atattaagct tggtaccgag
     6661 ctcggatcca tgggtaaggg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
     6721 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
     6781 acatacggaa aacttaccct taaatttatt tgcactactg gaaagcttcc tgttccttgg
     6841 ccaacacttg tcactactct tacttatggt gttcaatgct tttcaagata cccagatcat
     6901 atgaagcggc acgacttctt caagagcgcc atgcctgagg gatacgtgca ggagaggacc
     6961 atcttcttca aggacgacgg gaactacaag acacgtgctg aagtcaagtt tgagggagac
     7021 accctcgtca acagaatcga gcttaaggga atcgatttca aggaggacgg aaacatcctc
     7081 ggccacaagt tggaatacaa ctacaactcc cacaacgtat acatcatggc agacaaacaa
     7141 aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
     7201 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     7261 aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac
     7321 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
     7381 aaataagaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg
     7441 ccgcatcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
     7501 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
     7561 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
     7621 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
     7681 ggctttaatt tgcggcc
//
