LOCUS       62056_18480        4306 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 4306)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             1265..1981
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2033..2050
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2130..2376
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2391..2802
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2810..2857
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2876..3247
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3316..3563
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3638..4226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt
      961 tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga
     1021 agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga
     1081 tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa
     1141 tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc
     1201 tgaagctgaa ttcacgtggc ccagccggcc gtctcggatc ggtacctcga gccgcggcgg
     1261 ccgcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1321 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1381 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1441 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1501 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1561 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1621 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1681 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1741 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1801 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1861 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1921 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1981 gggtctagaa caaaaactca tctcagaaga ggatctgaat agcgccgtcg accatcatca
     2041 tcatcatcat tgagtttgta gccttagaca tgactgttcc tcagttcaag ttgggcactt
     2101 acgagaagac cggtcttgct agattctaat caagaggatg tcagaatgcc atttgcctga
     2161 gagatgcagg cttcattttt gatacttttt tatttgtaac ctatatagta taggattttt
     2221 tttgtcattt tgtttcttct cgtacgagct tgctcctgat cagcctatct cgcagctgat
     2281 gaatatcttg tggtaggggt ttgggaaaat cattcgagtt tgatgttttt cttggtattt
     2341 cccactcctc ttcagagtac agaagattaa gtgagacctt cgtttgtgcg gatcccccac
     2401 acaccatagc ttcaaaatgt ttctactcct tttttactct tccagatttt ctcggactcc
     2461 gcgcatcgcc gtaccacttc aaaacaccca agcacagcat actaaatttt ccctctttct
     2521 tcctctaggg tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa agagaccgcc
     2581 tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg
     2641 aaattttttt ttttagtttt tttctctttc agtgacctcc attgatattt aagttaataa
     2701 acggtcttca atttctcaag tttcagtttc atttttcttg ttctattaca acttttttta
     2761 cttcttgttc attagaaaga aagcatagca atctaatcta aggggcggtg ttgacaatta
     2821 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     2881 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     2941 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     3001 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     3061 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     3121 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     3181 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     3241 gcaggactga cacgtccgac ggcggcccac gggtcccagg cctcggagat ccgtccccct
     3301 tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc cctcccccca
     3361 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     3421 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     3481 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     3541 gacgctcgaa ggctttaatt tgcaagctgg agaccaacat gtgagcaaaa ggccagcaaa
     3601 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3661 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3721 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3781 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     3841 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3901 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3961 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4021 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4081 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4141 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4201 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4261 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagatc
//
