LOCUS       .                      10931 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        2..937
                     /label="AOX1 promoter"
                     /note="inducible promoter, regulated by methanol"
     CDS             949..1215
                     /label="α-factor secretion signal"
                     /note="N-terminal secretion signal from S. cerevisiae
                     alpha-factor"
     CDS             2935..2952
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      2976..3222
                     /label="AOX1 terminator"
                     /note="transcription terminator for AOX1"
     CDS             complement(6586..7398)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     misc_feature    7777..8533
                     /label="AOX1 3' fragment"
                     /note="region downstream of Pichia pastoris AOX1 gene"
     misc_feature    8676..8816
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9002..9590)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(9764..10621)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(10622..10726)
                     /label="AmpR promoter"
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 aaagtcgcca 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 tccaaacgat 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 agaattcatg tgcaaaggta ctttttattt aggtcaagaa
     1261 ggttcttctc tagaaagggt taatggttta caatctgcag aagttgaaga attgagacat
     1321 ttgagacaaa aaatggaaca agctcaggcc gatgtcggaa gtaaaaataa agaagtacaa
     1381 gcattacata ccaaattaaa tgctatggca caacaatatc aagatcaaca agctcatatt
     1441 tcattattaa aagaatcctt aaaagctaaa gaacagcata ttaatatgtt gcaagctgat
     1501 atgaattcat taagatccag aattgaagaa aaagaaatga ttcttgccca tcaagatgaa
     1561 cagttgaaca atctgtcctc tgataaatct aagagatccg gtgaggtgaa tgaactaaag
     1621 gaaattatag atttaaaaga aagaaaggtt gcagtgttac aaaaaaaaat tgataacctg
     1681 caagagcagt tgcaagaaaa aaaccatcag ttggaagatt tgagaggcaa agttaagtca
     1741 ttggaagctg attccaagtc atcggataat gcaatgactt ctttagaaga agcattgatg
     1801 gagaaagaca aatctctaat gttgttgaga gaacagaaag aaaaagaagg tgtttctttg
     1861 agagaagatt ctgaaaaaca tcaaagacaa actcatgaat tgacagctag aattgatgcc
     1921 ttacaaaagg atgtgaatga taaagaatct ggtctactag atttgaagga acatgcctct
     1981 caacttgcct ccgcaggttt gaaaaaagat agtaagataa ctacacttga aattaatttg
     2041 gcacaaacta aagaagaatt gtgtcaagct gaggaagaat taaagaaatg gaaagatgaa
     2101 aaggaagccg taacaggtca acaagtcaaa gaattgaacg aaaaagcaga aaaatcaaat
     2161 caagaggctc aaaaattgca atcagaagtt gatagaatat tagaaatcat gaaagacatg
     2221 gaaggagaaa aaaatgataa agataataga attaagacac tggagggtca attaaaggag
     2281 gctaacttga aattcacctc tatcaaaaaa gatcaacaaa atcaaaagaa ggcaaatcaa
     2341 caattactag aagaggctaa aaaaagagaa ggtgaactga ctaccgatac acatcaatta
     2401 caatctatgg ttaaagaaaa atcaaatcgg ttagaagaat tagaagaagc attgaaggaa
     2461 tcagttagca ttactgccga tagagaaatg attttagcac aacaaactca aaccgttaca
     2521 aaacttcaaa aacagttgga tgaaactaaa agaactttgc aagctgttca aaagaagcta
     2581 gcagataatg aaaacaaact aacagctact caagccattt tggaagaaaa ggataacaga
     2641 ttaaaaattt tggcagcgga aggaagaaaa catttagaag aagccttaga gacaaagcaa
     2701 gaagctttaa cagctgcaat cggcgaaaag gacgctcata tagctttact tgaattaaat
     2761 aggaaaggtt ctaagaccag tgatgaattg aaatctttga aaaaagaaaa ggacgctttg
     2821 gttcagcaat tgaaagatga aacacaaaaa agattgaaat tgcttcatca ttcctctgat
     2881 tcagataagt tgttaagagg taattcccct attacccctg aacaaagcgg ccgccaccat
     2941 caccatcacc attaaccggt cttgctagat tctaatcaag aggatgtcag aatgccattt
     3001 gcctgagaga tgcaggcttc atttttgata cttttttatt tgtaacctat atagtatagg
     3061 attttttttg tcattttgtt tcttctcgta cgagcttgct cctgatcagc ctatctcgca
     3121 gctgatgaat atcttgtggt aggggtttgg gaaaatcatt cgagtttgat gtttttcttg
     3181 gtatttccca ctcctcttca gagtacagaa gattaagtga gaagttcgtt tgtgcaagct
     3241 tatcgataag ctttaatgcg gtagtttatc acagttaaat tgctaacgca gtcaggcacc
     3301 gtgtatgaaa tctaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt
     3361 aggcataggc ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga
     3421 cagcatcgcc agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg
     3481 cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc
     3541 gctacttgga gccactatcg actacgcgat catggcgacc acacccgtcc tgtggatcta
     3601 tcgaatctaa atgtaagtta aaatctctaa ataattaaat aagtcccagt ttctccatac
     3661 gaaccttaac agcattgcgg tgagcatcta gaccttcaac agcagccaga tccatcactg
     3721 cttggccaat atgtttcagt ccctcaggag ttacgtcttg tgaagtgatg aacttctgga
     3781 aggttgcagt gttaactccg ctgtattgac gggcatatcc gtacgttggc aaagtgtggt
     3841 tggtaccgga ggagtaatct ccacaactct ctggagagta ggcaccaaca aacacagatc
     3901 cagcgtgttg tacttgatca acataagaag aagcattctc gatttgcagg atcaagtgtt
     3961 caggagcgta ctgattggac atttccaaag cctgctcgta ggttgcaacc gatagggttg
     4021 tagagtgtgc aatacacttg cgtacaattt caacccttgg caactgcaca gcttggttgt
     4081 gaacagcatc ttcaattctg gcaagctcct tgtctgtcat atcgacagcc aacagaatca
     4141 cctgggaatc aataccatgt tcagcttgag acagaaggtc tgaggcaacg aaatctggat
     4201 cagcgtattt atcagcaata actagaactt cagaaggccc agcaggcatg tcaatactac
     4261 acagggctga tgtgtcattt tgaaccatca tcttggcagc agtaacgaac tggtttcctg
     4321 gaccaaatat tttgtcacac ttaggaacag tttctgttcc gtaagccata gcagctactg
     4381 cctgggcgcc tcctgctagc acgatacact tagcaccaac cttgtgggca acgtagatga
     4441 cttctggggt aagggtacca tccttcttag gtggagatgc aaaaacaatt tctttgcaac
     4501 cagcaacttt ggcaggaaca cccagcatca gggaagtgga aggcagaatt gcggttccac
     4561 caggaatata gaggccaact ttctcaatag gtcttgcaaa acgagagcag actacaccag
     4621 ggcaagtctc aacttgcaac gtctccgtta gttgagcttc atggaatttc ctgacgttat
     4681 ctatagagag atcaatggct ctcttaacgt tatctggcaa ttgcataagt tcctctggga
     4741 aaggagcttc taacacaggt gtcttcaaag cgactccatc aaacttggca gttagttcta
     4801 aaagggcttt gtcaccattt tgacgaacat tgtcgacaat tggtttgact aattccataa
     4861 tctgttccgt tttctggata ggacgacgaa gggcatcttc aatttcttgt gaggaggcct
     4921 tagaaacgtc aattttgcac aattcaatac gaccttcaga agggacttct ttaggtttgg
     4981 attcttcttt aggttgttcc ttggtgtatc ctggcttggc atctcctttc cttctagtga
     5041 cctttaggga cttcatatcc aggtttctct ccacctcgtc caacgtcaca ccgtacttgg
     5101 cacatctaac taatgcaaaa taaaataagt cagcacattc ccaggctata tcttccttgg
     5161 atttagcttc tgcaagttca tcagcttcct ccctaatttt agcgttcaac aaaacttcgt
     5221 cgtcaaataa ccgtttggta taagaacctt ctggagcatt gctcttacga tcccacaagg
     5281 tggcttccat ggctctaaga ccctttgatt ggccaaaaca ggaagtgcgt tccaagtgac
     5341 agaaaccaac acctgtttgt tcaaccacaa atttcaagca gtctccatca caatccaatt
     5401 cgatacccag caacttttga gttgctccag atgtagcacc tttataccac aaaccgtgac
     5461 gacgagattg gtagactcca gtttgtgtcc ttatagcctc cggaatagac tttttggacg
     5521 agtacaccag gcccaacgag taattagaag agtcagccac caaagtagtg aatagaccat
     5581 cggggcggtc agtagtcaaa gacgccaaca aaatttcact gacagggaac tttttgacat
     5641 cttcagaaag ttcgtattca gtagtcaatt gccgagcatc aataatgggg attataccag
     5701 aagcaacagt ggaagtcaca tctaccaact ttgcggtctc agaaaaagca taaacagttc
     5761 tactaccgcc attagtgaaa cttttcaaat cgcccagtgg agaagaaaaa ggcacagcga
     5821 tactagcatt agcgggcaag gatgcaactt tatcaaccag ggtcctatag ataaccctag
     5881 cgcctgggat catcctttgg acaactcttt ctgccaaatc taggtccaaa atcacttcat
     5941 tgataccatt attgtacaac ttgagcaagt tgtcgatcag ctcctcaaat tggtcctctg
     6001 taacggatga ctcaacttgc acattaactt gaagctcagt cgattgagtg aacttgatca
     6061 ggttgtgcag ctggtcagca gcatagggaa acacggcttt tcctaccaaa ctcaaggaat
     6121 tatcaaactc tgcaacactt gcgtatgcag gtagcaaggg aaatgtcata cttgaagtcg
     6181 gacagtgagt gtagtcttga gaaattctga agccgtattt ttattatcag tgagtcagtc
     6241 atcaggagat cctctacgcc ggacgcatcg tggccgacct gcaggggggg ggggggcgct
     6301 gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc gccccatcat
     6361 ccagccagaa agtgagggag ccacggttga tgagagcttt gttgtaggtg gaccagttgg
     6421 tgattttgaa cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct
     6481 gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag ccgccgtccc gtcaagtcag
     6541 cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa actcatcgag
     6601 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     6661 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     6721 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     6781 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     6841 caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     6901 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     6961 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     7021 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     7081 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     7141 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     7201 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     7261 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     7321 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgagc aagacgtttc
     7381 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     7441 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac
     7501 gtggctttcc cccccccccc tgcaggtcgg catcaccggc gccacaggtg cggttgctgg
     7561 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     7621 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     7681 ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
     7741 ctgcttccta atgcaggagt cgcataaggg agagcgtcga gtatctatga ttggaagtat
     7801 gggaatggtg atacccgcat tcttcagtgt cttgaggtct cctatcagat tatgcccaac
     7861 taaagcaacc ggaggaggag atttcatggt aaatttctct gacttttggt catcagtaga
     7921 ctcgaactgt gagactatct cggttatgac agcagaaatg tccttcttgg agacagtaaa
     7981 tgaagtccca ccaataaaga aatccttgtt atcaggaaca aacttcttgt ttcgaacttt
     8041 ttcggtgcct tgaactataa aatgtagagt ggatatgtcg ggtaggaatg gagcgggcaa
     8101 atgcttacct tctggacctt caagaggtat gtagggtttg tagatactga tgccaacttc
     8161 agtgacaacg ttgctatttc gttcaaacca ttccgaatcc agagaaatca aagttgtttg
     8221 tctactattg atccaagcca gtgcggtctt gaaactgaca atagtgtgct cgtgttttga
     8281 ggtcatcttt gtatgaataa atctagtctt tgatctaaat aatcttgacg agccaaggcg
     8341 ataaataccc aaatctaaaa ctcttttaaa acgttaaaag gacaagtatg tctgcctgta
     8401 ttaaacccca aatcagctcg tagtctgatc ctcatcaact tgaggggcac tatcttgttt
     8461 tagagaaatt tgcggagatg cgatatcgag aaaaaggtac gctgatttta aacgtgaaat
     8521 ttatctcaag atctctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     8581 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     8641 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     8701 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     8761 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     8821 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     8881 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     8941 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     9001 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     9061 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     9121 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     9181 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     9241 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     9301 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     9361 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     9421 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     9481 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     9541 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     9601 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     9661 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     9721 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     9781 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     9841 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     9901 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     9961 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    10021 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
    10081 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    10141 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    10201 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    10261 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    10321 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
    10381 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    10441 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    10501 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    10561 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    10621 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
    10681 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
    10741 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
    10801 gtatcacgag gccctttcgt cttcaagaat taattctcat gtttgacagc ttatcatcga
    10861 taagctgact catgttggta ttgtgaaata gacgcagatc gggaacactg aaaaataaca
    10921 gttattattc g
//
