LOCUS       V014320                 7433 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014320
VERSION     V014320
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7433)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..937
                     /label="AOX1 promoter"
                     /note="inducible promoter, regulated by methanol"
     CDS             949..1215
                     /label="alpha-factor secretion signal"
                     /note="N-terminal secretion signal from S. cerevisiae
                     alpha-factor"
     CDS             1228..1257
                     /note="Laccase-2 from Cerrena unicolor. Accession#: P85149"
                     /label="Laccase-2"
     CDS             2734..2763
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             2779..2796
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      2876..3122
                     /label="AOX1 terminator"
                     /note="transcription terminator for AOX1"
     CDS             3265..4077
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     terminator      4153..4400
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4446..4462)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        4936..5040
                     /label="AmpR promoter"
     CDS             5041..5898
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      6072..6660
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    6948..6969
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6984..7014
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    7022..7038
                     /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     7046..7062
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
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 attcgaagga tccaacatat gagatttcct
      961 tcaattttta ctgcagtttt attcgcagca tcctccgcat tagctgctcc agtcaacact
     1021 acaacagaag atgaaacggc acaaattccg gctgaagctg tcatcggtta ctcagattta
     1081 gaaggggatt tcgatgttgc tgttttgcca ttttccaaca gcacaaataa cgggttattg
     1141 tttataaata ctactattgc cagcattgct gctaaagaag aaggggtatc tctcgagaaa
     1201 agagaggctg aagcttacgt agaattcgct attggccctg tagctgatct ccacatttcc
     1261 aatgctaaca tctctcccga tggcttcact cgtgcagccg ttctcgctgg cgggtccttc
     1321 cctggacctc ttattactgg caataagggt gacaacttcc agatcaacgt gatcaacgac
     1381 ctcaccgatg cagaccaact caagaccacc accattcact ggcatggttt cttccagcac
     1441 ggtactaact gggctgacgg cccggctttc atcaatcagt gtcctattgc ctccggcaac
     1501 tcgttcttgt acaacttcca agttcctgac caagctggta cattctggta tcacagccat
     1561 ctctcgactc agtattgtga cggactccgc ggagcattcg tggtctatga tcccgatgac
     1621 ccacatgcaa gtttgtacga cgtcgatgat gagagtaccg tcatcactct tgcagactgg
     1681 taccatgacc tagcgcggtt gggtccgaag ttccccacca ctaactcgac actcattaac
     1741 ggccttggtc gttacgattt cggaccctcc tccgacttgg ctgtcatctc cgtgcaagct
     1801 ggaaagcggt accgtttccg tctggtctct atctcgtgtg actcgaacta cgtgttctcc
     1861 atcgacgagc acacattcac cgtcatcgaa gtcgacggtg tcaaccacca acctgtcgag
     1921 gctgattccc tccaaatctt ccctggtcag cggtactcca tcgtggtcaa tgccgacaaa
     1981 tcggaaggag gtaactattg gattcgcgcc tcacctactc ctggtcccca aggatttgcc
     2041 aatggaatca actctgctat cttgcgctat gtcggttccg atgaggttga gcctactacg
     2101 aaccagacga cgagcacaaa cccgcttgtg gaggctaacc tggttcccct tgagaaccct
     2161 ggagcgcctg gcgatcccac acccggcggt gtagatgtcc ccctcaacct tgcagtcgct
     2221 ttcgatggta caggactcga cttccaggtc aacggacaga cgttcatccc ccctaccgtt
     2281 cccgtcttgc tgcaaattct cagcggtgct caatctgcca cagacctcct ccccagcgga
     2341 agcgtctacg ttcttccgag caacgctacg gtcgaaataa gcattccagc tggcgccgtc
     2401 ggcggtcctc accccattca tttgcacggt cacacattcg atgtcgtccg aagtgccggc
     2461 agcagcactt ataactatgt caacccgcct cgtcgcgacg tcgtcagcat tggtaacgcc
     2521 ggtgacaacg taaccattcg tttccggact gataaccctg gaccatggtt cctccattgc
     2581 cacatcgact ggcatctcga agccggcttt gctgttgtct tcgctgaaga catccccaac
     2641 gtcgcctcag tcaactcccc tcctcaggcc tggagcgacc tttgccctat ctatgatgcg
     2701 cttgaccctt ctgaccactg agcggccgct ctagaacaaa aactcatctc agaagaggat
     2761 ctgaatagcg ccgtcgacca tcatcatcat catcattgag tttgtagcct tagacatgac
     2821 tgttcctcag ttcaagttgg gcacttacga gaagaccggt cttgctagat tctaatcaag
     2881 aggatgtcag aatgtcattt gcctgagaga tgcaggcttc atttttgata cttttttatt
     2941 tgtaacctat atagtatagg attttttttg tcattttgtt tcttctcgta cgagcttgct
     3001 cctgatcagc ctatctcgca gctgatgaat atcttgtggt aggggtttgg gaaaatcatt
     3061 cgagtttgat gtttttcttg gtatttccca ctcctcttca gaatacagaa gattaagtga
     3121 gaccttcgtt tgtgcggatc cgctagctta gtgtactacc caatcttgcc tcttcgggtg
     3181 tcttttataa ggacagattc acaagctctt gttgcccaat acacacatac acacagagat
     3241 aatagcagtc ttaattaaat gaaaatgagc catattcaac gggaaacgtc ttgctcaagg
     3301 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     3361 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
     3421 tttctgaaac atggcaaagg tagcgttgct aatgatgtta cagatgagat ggtcagacta
     3481 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     3541 gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa
     3601 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     3661 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     3721 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     3781 tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca
     3841 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     3901 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     3961 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     4021 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     4081 agatctcacg tccgacgggc ggcccacggg tcccaggcct cggagatccg tccccctttt
     4141 cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat
     4201 ccgctctaac caaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     4261 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     4321 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac
     4381 gctcgaaggc tttaatttgc aagctagtac ttttaatcgt ccacttgccg tcatgcttct
     4441 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4501 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     4561 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     4621 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     4681 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     4741 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4801 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     4861 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4921 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4981 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5041 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5101 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5161 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5221 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5281 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5341 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5401 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5461 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5521 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5581 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5641 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5701 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5761 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5821 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5881 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5941 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6001 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6061 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6181 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6721 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6781 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6841 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     6901 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     6961 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7021 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgaactcgtc
     7081 ctcggttccc ggcgatcctc tggagattgc ttaccaaaaa tggcccactg agggggtcta
     7141 gtatttgaag aagcctcact aaagcaggtc ttcgccacac atttaaagtt tgagaatggg
     7201 cggaggtcaa ccccccgtcc ccctaatcat tgctttacct catggaacta aacaaccacc
     7261 cccccgctat cctgagggaa acttcggcag gaaccagcta ctagatggtt cgattagtct
     7321 ttcgccccta tacccaaatt cgacgatcga tttgcacgtc agaaccgctg cgggcctcca
     7381 ccagagtttc ctcaggcttc gccctattca ggcatagttc accatctact agt
//
