LOCUS       sequence-256438-       15179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid overexpressing ACC1 from Komagataella phaffii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15179)
  AUTHORS   Liu Y, Bai C, Liu Q, Xu Q, Qian Z, Peng Q, Yu J, Xu M, Zhou X, Zhang
            Y, Cai M
  TITLE     Engineered ethanol-driven biosynthetic system for improving 
            production of acetyl-CoA derived drugs in Crabtree-negative yeast.
  JOURNAL   Metab Eng. 2019 Jul;54:275-284. doi: 10.1016/j.ymben.2019.05.001. 
            Epub 2019 May 9.
  PUBMED    31077813
REFERENCE   2  (bases 1 to 15179)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab 
            Eng."; date: "2019-07"; volume: "54:275-284. doi"; pages: " 
            10.1016/j.ymben.2019.05.001. Epub 2019 May 9"
COMMENT     Sequence Label: pK_P-GAP-ACC1*
FEATURES             Location/Qualifiers
     source          1..15179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        143..618
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     regulatory      729..738
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3603..3614)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             7264..7281
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      7361..7607
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(8026..10560)
                     /codon_start=1
                     /gene="Pichia pastoris HIS4"
                     /product="multifunctional enzyme, required for histidine 
                     biosynthesis"
                     /label=PpHIS4
                     /note="auxotrophic marker for Pichia pastoris"
                     /translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
                     LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
                     VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
                     DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
                     PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
                     TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
                     IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
                     KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
                     LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
                     GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
                     VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
                     LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
                     AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
                     QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
                     HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
     CDS             complement(10974..11789)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(10974..11789)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     11697..11716
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     11781..11800
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(11954..11973)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_feature    12168..12916
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     primer_bind     complement(12952..12974)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13060..13200
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     13115..13134
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(13215..13232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13386..13974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13466..13485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14145..14936,14937..15005))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     14768..14787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15006..15110)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(15178..15179,1..17)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 taggcgtatc acgaggccct ttcgtcttca agaattaatt ctcatgtttg acagcttatc
       61 atcgataagc tgactcatgt tggtattgtg aaatagacgc agatcgggaa cactgaaaaa
      121 taacagttat tattcggagc tctttttgta gaaatgtctt ggtgtcctcg accaatcagg
      181 tagccatccc tgaaatacct ggctccgtgg caacaccgaa cgacctgctg gcaacgttaa
      241 attctccggg gtaaaactta aatgtggagt aatagaacca gaaacgtctc ttcccttctc
      301 tctccttcca ccgcccgtta ccgtccctag gaaattttac tctgctggag agcttcttct
      361 acggccccct tgcagcaatg ctcttcccag cattacgttg cgggtaaaac ggaggtcgtg
      421 tacccgacct agcagcccag ggatggaaag tcccggccgt cgctggcaat aactgcgggc
      481 ggacgcatgt cttgagatta ttggaaacca ccagaatcga atataaaagg cgaacacctt
      541 tcccaatttt ggtttctcct gacccaaaga ctttaaattt aatttatttg tccctatttc
      601 aatcaattga acaactatat gagtagtgtt aaccactctc tccgtcattc aaagctaccg
      661 ccgcatttcc ttggtctcaa ctcggttgaa gtcgctgctc cctccaaggt cagagacttt
      721 gtcagggacc atggtggcca ctcggtcatc acgagagtgc tgatcgcaaa caacggtata
      781 gctgccgtga aagaaattcg ttccgtcagg aaatgggcgt atgaaacgtt tggtaacgat
      841 agagccattc aatttattgt tatggctacc ccagaggatc ttgaagctaa tgctgaatat
      901 attcgaatgg ctgaccagta tgtcatggtc ccaggaggaa ctgcaaacaa caactatgcg
      961 aacgtcgacc tcattgtaga aatagcagaa tctactgatg ctcatgctgt ttgggctggt
     1021 tggggttttg cctccgaaaa tccccatttg cctgagcaac tggccgcttc tcctaagaag
     1081 attatcttca ttggccctcc gggctctgcc atgcgatctc ttggtgacaa gatttcctct
     1141 actattgtcg cacaacatgc taaagtccca tgtattcctt ggtcaggaac tggtgtcgat
     1201 caggttataa tcgaccccgt aagcaatttg gtttccgttg atgaagaaac gtacgccaaa
     1261 ggatgctgtt ccgatccaca ggacggtttg gcaaaagcca aggctattgg tttccctgtg
     1321 atgattaaag cttccgaagg tggtggtggt aaaggaatta gaaaagttga cagggaggaa
     1381 gattttcttt ctctttatga tcaagctgct aatgaaattc caggttcccc aatttttatc
     1441 atgaagcttg ctggagatgc caggcatttg gaagttcaat tacttgctga tcaatatgga
     1501 accaacatct ccctttttgg aagagattgt tccgttcaaa gaagacacca aaagatcata
     1561 gaagaggcac cagttaccat tgccaaacaa gacactttca ggcaaatgga acaagccgct
     1621 gtcagactgg gtcaattggt tggatacgtt tctgccggta ccgttgagta tctatattca
     1681 cacgctgagg acaagttcta cttcttggaa ctgaaccctc gtcttcaagt tgagcatcca
     1741 accacagaaa tggccacagg tgtcaatctt ccagttgccc agttgctaat tgcaatgggt
     1801 attcctttga atagaatcag agatatcagg gtactttacg gacttgaacc aaatggcgct
     1861 acagaaattg actttgaatt caaaactgaa gaaagcttga agagtcaaag aaaacccatt
     1921 ccaaagggtc acactattgc atgtcgtatc acatctgaag atcctggtga aggttttaag
     1981 ccttctggtg gtgctctata tgagctaaat ttcagatctt cttctagcgt ttggggttac
     2041 ttcagtgtag gaaacaaatc ctcaattcat tctttcagtg actctcaatt tggtcatata
     2101 ttctcgtttg gcgaaaaccg tcaaatcgcc agaaaaaata tggtcgtcgc cttgaaagag
     2161 ctttctattc gtggtgactt tagaactaca attgagtact taataaaact gttggaaaca
     2221 gctgatttcg agaacaacac catcactact ggttggttgg acgaactgat ctcgaagaag
     2281 ctgactgctg aaagacctga tgaaaccaca gcaattttat gtggtgctga aaaaggtcaa
     2341 atcccaggca aagaacttct tcgtactatt ttcccaattg aatttattta tgaaggaaag
     2401 aagtacaagt ttactgtggt tcaggctgca tttgacaaat acaacgtctt tgtcaacgga
     2461 tgtatgatta ctgtaagtgt aacccatttg aaggatggca gtttattggt agcacttgat
     2521 ggtaaatccc attctgtcta ttacttgcag gaagaagtcg gaaatactag gttgtcggtg
     2581 gatggtaaat cttgcatttt agaagttgag catgagccaa ctgaacttcg tactccatct
     2641 ccaggtaaac ttatcaaata tcttgtggaa cacggtgatc acgtcaaaat tggacaacct
     2701 tacgctgaag ttgaagtaat gaagatgtgt atgcctttgg tcagtcagga gaatggaact
     2761 atcaggttat tgaagcagcc aggatcttcg gttgccgctg gagacatcct tgctattctt
     2821 gcattggatg atcccagcaa ggtgaagcat gctttgccat tcgatggtac aatccctgat
     2881 atgaaacagc catttatcca tagcaacaaa ccagtttata agttcatttc tcttctctcc
     2941 gtgctgaaaa acattttagc agggtatgat aatcaagttg tgatgaacga tactctgcag
     3001 agtctattgg atgtgttgaa gaaccctgaa cttccttatt cggaatggaa tcattcgata
     3061 tctgcacttc attcaaggtt accaattcat ttggacgaac aattgaccag tttgattgag
     3121 agatcgcatc aacgtggtgc agactttcca gctaagcact tgctcaagct tttggacaag
     3181 gagcaggctg ttaatcctga tccacttttc tcccaggtca ttgcgcctct tactgctgtt
     3241 gccaaaagct acgaacatgg acttgaagtt catgaacaca atgtattcgc cgatttgatc
     3301 acccaatact acgacataga gagcttgttt gccgataaaa gggaggaaga tgttatttta
     3361 cagctacgtg atgagaacaa atcgtccctt gacaaggtca tcgatgtcgt cttgtcacat
     3421 tccagagttg gagctaagaa ccatttaatc agagctattc tggaaattta tcaaactatc
     3481 tgccaaaatg atctccaagc tgcaaccatt ttgaagaaac ctttgaaaaa gattgttgag
     3541 ctagattcta gatttacagc aaaggtttcg ttaaaagcta gagagatttt gattcaatgt
     3601 tcccttccct ctatcaaaga acgttcagac cagctcgagc atatccttcg atcttcagtt
     3661 gtacaaactc agtacggaga gagcttcaat ggaaactaca aactgcctaa cttggacgtt
     3721 atacaagacg taattgattc caagtacatt gtattcgatg ttttgacaca atttgttgtt
     3781 agcccaaaca agtatatatt tgcagcagca gccgaggtgt atctgcgaag agcttacagg
     3841 gcttactcgg tgagagaagt taaacatcat ttcgtaggtg attctgctct cccaattgtg
     3901 gaatggaagt tccaattgcc gctgttatca acagctgctt acaattccgt gcctgaagct
     3961 atgagaaact cctccagtaa ccgatcctct atttcaatgg atagagcagt tgctgtctcc
     4021 gatttgacct tcatgatcaa caagaatgat tctcaacctt tgagaacagg tatcataatt
     4081 cccacaaacc acttagatga cattgaggag tccttgtcat ctgccattga tgtcttccct
     4141 aaacgtccac gtaacaatgg accagctcct gacagaacta atgtggctcc tgagcaacct
     4201 actaacgtat gcaatgtttt cattgccaat gtttctggct acaacagtga ggctgagatc
     4261 gttgacaaga ttagcagcgt tctttctgag ttgaaagacg acctcagggc tagtggcgtt
     4321 cgaagagtta cctttgtctt gggagacaag gttggaactt atccaaaata ctataccttc
     4381 aaatttccag actattttga agacgagaca atccgtcaca tagagcctgc tcttgcgttc
     4441 cagctggaac taagaagatt gtccaatttc aatattaaac ctgttccaac tgagaataga
     4501 aatattcatg tgtatgaggc agttgccaaa aatacttcat gcattgacag gagatttttt
     4561 actaggggta tcatcagaac aagcagaatc agagaggatg tgactatctc tgaatacctt
     4621 atcagcgaag ctaatcgtct tatgagtgac attttggacg ctcttgagat tattgatacc
     4681 tccaacactg atttgaacca tatattcatc aatttctctg ctgttttcaa tgtcacgcca
     4741 gatgacgttg aagcagcgtt cggtggtttc ttagaaaggt ttggacgtag gctgtggaga
     4801 ctacgtgttt ctgctgctga aatccgtatt atgtgcacgg accctgagac tggtatccca
     4861 ttcccacttc gtgctttaat taacaacgtt tcaggatacg ttgtgaaatc tgaaatgtat
     4921 caagaggtga aaaatgatca tggggaatgg gttttcaaaa gtcttggtcc tacaccaggt
     4981 tcaatgcacc ttagaccaat ttcaacacca tacccaacca aagaatggct tcaaccaaaa
     5041 cgttacaaag ctcatcttat gggtactact tacgtgtatg atttccctga attattccgt
     5101 caagctacgc tctcccaatg gaaaaaatac tctcctactg cgagagttcc ttctgatgtg
     5161 tttgtggcca atgaattgat cgtcgatgat tcaggtgaac taactgaagt aagcagagaa
     5221 cccggcgcca acgttgtggg tatggtggcc ttcaaggtaa ccgcaaaaac tcctgagtat
     5281 ccacgcggtc gccatttcat cataattgct aatgatatca ccttcaagat cggatccttt
     5341 ggccctcaag aagatgaata tttcaacaag gccacacaac ttgcaagaaa attgggcatt
     5401 cctcgaattt atctgtcagc caactcgggt gctagaattg gagttgctga agaacttctt
     5461 ccattattca aagtagcctg gaaggaagaa ggtaaaccaa gcaagggatt tgaatactta
     5521 tacctcacat cggaagatct tactctattg gaaaagtccg gaaagtctaa cagcgttacc
     5581 actcaaagaa tagttgaaga aggcgaagaa cgccacgtta taactgccat cattggagct
     5641 agtgatggac tgggtgttga atgtctaaga ggttccggtt tgatcgctgg tgctacatct
     5701 cgggcgtaca aggacatctt cactatcaca ttggtcacct gtagatctgt tggtattggt
     5761 gcttacttgg tcagattggg tcaacgagcc attcaaattg aaggacaacc aataattttg
     5821 actggtgccc ctgctattaa taagttgttg ggtagggaag tgtactcttc caacctgcaa
     5881 cttggtggta cccagattat gtacaagaac ggtgtttcac acttaaccgc caatgatgat
     5941 ctcgcaggtg tcgaaaagat tatggattgg ttagcttatg tgcctgctaa gagaaacatg
     6001 cctgttccta ttttagaatc acttcatgac aaatgggaca gagatgtgga ctataagcct
     6061 acaagaaatg agccgtacga cgtcagatgg atgatcagtg gacgtgaaac tcctgatggt
     6121 gagttcgaat ctggattgtt tgactctggg tccttcactg aaactttgag tggatgggct
     6181 aaaggtgtag tcgtcggaag agcccgttta ggtggtattc ctatgggagt cattggtgtt
     6241 gaaactagag tcacagaaaa cctgattcca gctgatcccg ccaatccaga ctcaaccgaa
     6301 atgatgattc aagaagctgg tcaagtctgg taccctaaca gtgccttcaa gactgcacaa
     6361 gctatcaacg atttcaacaa tggtgaacag ctacccttga tgattttggc caactggaga
     6421 ggtttctctg gtggtcaaag agacatgtac aatgaagttt tgaaatacgg ttctttcatt
     6481 gtggatgctt tagtcgactt caagcagcct atcttcactt acattcctcc cactgctgag
     6541 ttgagaggtg gatcttgggt tgttgtagac cctaccatca atgaagacat gatggaaatg
     6601 tatgcagacg tcgaatcaag agcaggtgtt ttggaaccag aaggtatggt aggtatcaaa
     6661 taccgtaagg acaaactcct tgctactatg gaacgattgg atgccaaata tgctgagctt
     6721 aaatccaagg ttagcgatac tagtctttca gaaaaggatg tttccgagat caagaaacaa
     6781 attgagcaga gagagaagca attgttgcca atttatgcac aaatctctat tcaatttgct
     6841 gatcttcatg acagatctgg tcgtatgttg gccaagggtg tcattaaaaa ggaactggaa
     6901 tgggttaatt ctcgtcgttt cttcttctgg agagtccgtc gtcgtttgaa cgaggaatac
     6961 ctcattaagc gtattaccga attcctatct gcttctgcta ccagattgga caagatctcg
     7021 aggatcaatt cttggttgcc aacatcgatt gatttggaag atgaccagaa ggttgccatt
     7081 tggttggaag aaaaccgtaa agctcttgac gccaatatca aggagctcag ggctgagcat
     7141 gttagaagaa ctctggctac tcttgtcaga actgatatgg atactacttc caagagtttg
     7201 gctgaattga tcaaccttct tcctgaaacc gaaaaggaat caattttatc taagatcaag
     7261 tcacatcatc atcatcatca ttgagtttgt agccttagac atgactgttc ctcagttcaa
     7321 gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc
     7381 catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt
     7441 ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc
     7501 tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt
     7561 tcttggtatt tcccactcct cttcagagta cagaagatta agtgagaact agtagttcgt
     7621 ttgtgcaagc ttatcgataa gctttaatgc ggtagtttat cacagttaaa ttgctaacgc
     7681 agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg caccgtcacc
     7741 ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt gcgggatatc
     7801 gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata tgcgttgatg
     7861 caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg ccgcccagtc
     7921 ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac cgcacccgtc
     7981 ctgtggatct atcgaatcta aatgtaagtt aaaatctcta aataattaaa taagtcccag
     8041 tttctccata cgaaccttaa cagcattgcg gtgagcatct agaccttcaa cagcagccag
     8101 atccatcact gcttggccaa tatgtttcag tccctcagga gttacgtctt gtgaagtgat
     8161 gaacttctgg aaggttgcag tgttaactcc gctgtattga cgggcatatc cgtacgttgg
     8221 caaagtgtgg ttggtaccgg aggagtaatc tccacaactc tctggagagt aggcaccaac
     8281 aaacacagat ccagcgtgtt gtacttgatc aacataagaa gaagcattct cgatttgcag
     8341 gatcaagtgt tcaggagcgt actgattgga catttccaaa gcctgctcgt aggttgcaac
     8401 cgatagggtt gtagagtgtg caatacactt gcgtacaatt tcaacccttg gcaactgcac
     8461 agcttggttg tgaacagcat cttcaattct ggcaagctcc ttgtctgtca tatcgacagc
     8521 caacagaatc acctgggaat caataccatg ttcagcttga gacagaaggt ctgaggcaac
     8581 gaaatctgga tcagcgtatt tatcagcaat aactagaact tcagaaggcc cagcaggcat
     8641 gtcaatacta cacagggctg atgtgtcatt ttgaaccatc atcttggcag cagtaacgaa
     8701 ctggtttcct ggaccaaata ttttgtcaca cttaggaaca gtttctgttc cgtaagccat
     8761 agcagctact gcctgggcgc ctcctgctag cacgatacac ttagcaccaa ccttgtgggc
     8821 aacgtagatg acttctgggg taagggtacc atccttctta ggtggagatg caaaaacaat
     8881 ttctttgcaa ccagcaactt tggcaggaac acccagcatc agggaagtgg aaggcagaat
     8941 tgcggttcca ccaggaatat agaggccaac tttctcaata ggtcttgcaa aacgagagca
     9001 gactacacca gggcaagtct caacttgcaa cgtctccgtt agttgagctt catggaattt
     9061 cctgacgtta tctatagaga gatcaatggc tctcttaacg ttatctggca attgcataag
     9121 ttcctctggg aaaggagctt ctaacacagg tgtcttcaaa gcgactccat caaacttggc
     9181 agttagttct aaaagggctt tgtcaccatt ttgacgaaca ttgtcgacaa ttggtttgac
     9241 taattccata atctgttccg ttttctggat aggacgacga agggcatctt caatttcttg
     9301 tgaggaggcc ttagaaacgt caattttgca caattcaata cgaccttcag aagggacttc
     9361 tttaggtttg gattcttctt taggttgttc cttggtgtat cctggcttgg catctccttt
     9421 ccttctagtg acctttaggg acttcatatc caggtttctc tccacctcgt ccaacgtcac
     9481 accgtacttg gcacatctaa ctaatgcaaa ataaaataag tcagcacatt cccaggctat
     9541 atcttccttg gatttagctt ctgcaagttc atcagcttcc tccctaattt tagcgttcaa
     9601 caaaacttcg tcgtcaaata accgtttggt ataagaacct tctggagcat tgctcttacg
     9661 atcccacaag gtggcttcca tggctctaag accctttgat tggccaaaac aggaagtgcg
     9721 ttccaagtga cagaaaccaa cacctgtttg ttcaaccaca aatttcaagc agtctccatc
     9781 acaatccaat tcgataccca gcaacttttg agttgctcca gatgtagcac ctttatacca
     9841 caaaccgtga cgacgagatt ggtagactcc agtttgtgtc cttatagcct ccggaataga
     9901 ctttttggac gagtacacca ggcccaacga gtaattagaa gagtcagcca ccaaagtagt
     9961 gaatagacca tcggggcggt cagtagtcaa agacgccaac aaaatttcac tgacagggaa
    10021 ctttttgaca tcttcagaaa gttcgtattc agtagtcaat tgccgagcat caataatggg
    10081 gattatacca gaagcaacag tggaagtcac atctaccaac tttgcggtct cagaaaaagc
    10141 ataaacagtt ctactaccgc cattagtgaa acttttcaaa tcgcccagtg gagaagaaaa
    10201 aggcacagcg atactagcat tagcgggcaa ggatgcaact ttatcaacca gggtcctata
    10261 gataacccta gcgcctggga tcatcctttg gacaactctt tctgccaaat ctaggtccaa
    10321 aatcacttca ttgataccat tattgtacaa cttgagcaag ttgtcgatca gctcctcaaa
    10381 ttggtcctct gtaacggatg actcaacttg cacattaact tgaagctcag tcgattgagt
    10441 gaacttgatc aggttgtgca gctggtcagc agcataggga aacacggctt ttcctaccaa
    10501 actcaaggaa ttatcaaact ctgcaacact tgcgtatgca ggtagcaagg gaaatgtcat
    10561 acttgaagtc ggacagtgag tgtagtcttg agaaattctg aagccgtatt tttattatca
    10621 gtgagtcagt catcaggaga tcctctacgc cggacgcatc gtggccgacc tgcanggggg
    10681 gggggggcgc tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc aggcctgaat
    10741 cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt tgttgtaggt
    10801 ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt tgtcgggaag
    10861 atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa gccgccgtcc
    10921 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa
    10981 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
    11041 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
    11101 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
    11161 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
    11221 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
    11281 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
    11341 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
    11401 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
    11461 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
    11521 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
    11581 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
    11641 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
    11701 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag
    11761 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
    11821 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
    11881 tgagacacaa cgtggctttc cccccccccc ctgcaggtcg gcatcaccgg cgccacaggt
    11941 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
    12001 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
    12061 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
    12121 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agtatctatg
    12181 attggaagta tgggaatggt gatacccgca ttcttcagtg tcttgaggtc tcctatcaga
    12241 ttatgcccaa ctaaagcaac cggaggagga gatttcatgg taaatttctc tgacttttgg
    12301 tcatcagtag actcgaactg tgagactatc tcggttatga cagcagaaat gtccttcttg
    12361 gagacagtaa atgaagtccc accaataaag aaatccttgt tatcaggaac aaacttcttg
    12421 tttcgaactt tttcggtgcc ttgaactata aaatgtagag tggatatgtc gggtaggaat
    12481 ggagcgggca aatgcttacc ttctggacct tcaagaggta tgtagggttt gtagatactg
    12541 atgccaactt cagtgacaac gttgctattt cgttcaaacc attccgaatc cagagaaatc
    12601 aaagttgttt gtctactatt gatccaagcc agtgcggtct tgaaactgac aatagtgtgc
    12661 tcgtgttttg aggtcatctt tgtatgaata aatctagtct ttgatctaaa taatcttgac
    12721 gagccagacg ataataccaa tctaaactct ttaaacgtta aaggacaagt atgtctgcct
    12781 gtattaaacc ccaaatcagc tcgtagtctg atcctcatca acttgagggg cactatcttg
    12841 ttttagagaa atttgcggag atgcgatatc gagaaaaagg tacgctgatt ttaaacgtga
    12901 aatttatctc aagatctgct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
    12961 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
    13021 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
    13081 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
    13141 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
    13201 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
    13261 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
    13321 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
    13381 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
    13441 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
    13501 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
    13561 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
    13621 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
    13681 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
    13741 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
    13801 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
    13861 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
    13921 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
    13981 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
    14041 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
    14101 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
    14161 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
    14221 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
    14281 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
    14341 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
    14401 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
    14461 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
    14521 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
    14581 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
    14641 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
    14701 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    14761 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    14821 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    14881 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    14941 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    15001 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
    15061 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
    15121 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaa
//
