LOCUS       62056_18500        4040 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 4040)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             1000..1716
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1768..1785
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1864..2110
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2125..2536
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2544..2591
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2610..2981
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3050..3297
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3372..3960)
                     /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 aggaattcac gtggcccagc
      961 cggccgtctc ggatcggtac ctcgagccgc ggcggccgca tggtgagcaa gggcgaggag
     1021 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1081 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1141 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1201 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1261 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1321 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1381 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1441 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1501 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1561 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     1621 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1681 gccgggatca ctctcggcat ggacgagctg tacaaggggc ccgaacaaaa actcatctca
     1741 gaagaggatc tgaatagcgc cgtcgaccat catcatcatc atcattgagt tttagcctta
     1801 gacatgactg ttcctcagtt caagttgggc acttacgaga agaccggtct tgctagattc
     1861 taatcaagag gatgtcagaa tgccatttgc ctgagagatg caggcttcat ttttgatact
     1921 tttttatttg taacctatat agtataggat tttttttgtc attttgtttc ttctcgtacg
     1981 agcttgctcc tgatcagcct atctcgcagc tgatgaatat cttgtggtag gggtttggga
     2041 aaatcattcg agtttgatgt ttttcttggt atttcccact cctcttcaga gtacagaaga
     2101 ttaagtgaga ccttcgtttg tgcggatccc ccacacacca tagcttcaaa atgtttctac
     2161 tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca
     2221 cccaagcaca gcatactaaa ttttccctct ttcttcctct agggtgtcgt taattacccg
     2281 tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag
     2341 gcaataaaaa tttttatcac gtttcttttt cttgaaattt ttttttttag tttttttctc
     2401 tttcagtgac ctccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag
     2461 tttcattttt cttgttctat tacaactttt tttacttctt gttcattaga aagaaagcat
     2521 agcaatctaa tctaaggggc ggtgttgaca attaatcatc ggcatagtat atcggcatag
     2581 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
     2641 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     2701 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     2761 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     2821 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     2881 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     2941 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtc cgacggcggc
     3001 ccacgggtcc caggcctcgg agatccgtcc cccttttcct ttgtcgatat catgtaatta
     3061 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     3121 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     3181 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     3241 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag
     3301 ctggagacca acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3361 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3421 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3481 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3541 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     3601 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3661 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3721 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3781 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     3841 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3901 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3961 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4021 gggattttgg tcatgagatc
//
