LOCUS       62056_18475        4189 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 4189)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             941..1207
                     /codon_start=1
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     CDS             1916..1933
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2013..2259
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2274..2685
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2693..2740
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2759..3130
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3199..3446
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3521..4109)
                     /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 tgaagctgcg acgggactcg acgacccggc gaagaaggag atcgccatgc aactggtgtc
     1261 gagcgccgag aactcctcgc tcgactggaa ggcccagtac cggtacatcg aggacatcgg
     1321 tgacggtcgc ggctacaccg cgggcatcat cggcttctgt tccggcaccg gcgacatgct
     1381 cgacctggtg gagctgtacg gcgagcgcag ccccgggaac gtcctcgcgc cctatctgcc
     1441 ggcgctgcgc cgggtggacg gctccgactc gcacgagggc ctcgaccccg gcttcccgga
     1501 cgactggcgc cgggccgccg accaggaccc gcagttccgc cgggcccagg acgacgagcg
     1561 ggaccgcgtg tacttcgacc cggccgtacg gcgcggcaag gaggacgggc tgcggacgct
     1621 ggggcagttc gcgtactacg acgcgatggt gatgcacggc gacggcggcg gcctcggctt
     1681 cggcagtatc cgcgagcgcg ccctcggccg ggcccgcccg ccggcccagg gcggtgacga
     1741 ggtcgcctac ctgcacgcct tcctcgacga gcgggtgtgg gccatgaagc aggagcaggc
     1801 gcacagcgac accagccggg tcgacacggc gcagcgcgtc ttcctgaacg agggcaacct
     1861 ggacctcgag ccgccgctgg actggcacgt gtacggggac gcctaccaca tcggccatca
     1921 tcatcatcat cattgagttt gtagccttag acatgactgt tcctcagttc aagttgggca
     1981 cttacgagaa gaccggtctt gctagattct aatcaagagg atgtcagaat gccatttgcc
     2041 tgagagatgc aggcttcatt tttgatactt ttttatttgt aacctatata gtataggatt
     2101 ttttttgtca ttttgtttct tctcgtacga gcttgctcct gatcagccta tctcgcagct
     2161 gatgaatatc ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt tttcttggta
     2221 tttcccactc ctcttcagag tacagaagat taagtgagac cttcgtttgt gcggatcccc
     2281 cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac
     2341 tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat tttccctctt
     2401 tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc
     2461 gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc
     2521 ttgaaatttt tttttttagt ttttttctct ttcagtgacc tccattgata tttaagttaa
     2581 taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
     2641 ttacttcttg ttcattagaa agaaagcata gcaatctaat ctaaggggcg gtgttgacaa
     2701 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     2761 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga
     2821 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
     2881 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa
     2941 caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt
     3001 cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc
     3061 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga
     3121 ggagcaggac tgacacgtcc gacggcggcc cacgggtccc aggcctcgga gatccgtccc
     3181 ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca cgccctcccc
     3241 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     3301 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     3361 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     3421 tgggacgctc gaaggcttta atttgcaagc tggagaccaa catgtgagca aaaggccagc
     3481 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3541 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3601 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3661 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     3721 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3781 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3841 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3901 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3961 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4021 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4081 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4141 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatc
//
