LOCUS       sequence-246578-       17306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid overexpressing sLovA,CPR and LacI-Mit1AD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17306)
  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 17306)
  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-ICL1-LM_lacO-cP-AOX1-sAR
FEATURES             Location/Qualifiers
     source          1..17306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    133..149
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             1922..1939
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2019..2265
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     protein_bind    2281..2297
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             4571..4588
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4668..4914
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             9373..9390
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      9470..9716
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(10135..12669)
                     /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(13082..13897)
                     /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(13082..13897)
                     /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     13805..13824
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     13889..13908
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(14062..14081)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_feature    14276..15024
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     primer_bind     complement(15060..15082)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    15168..15308
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     15223..15242
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(15323..15340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(15494..16082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15574..15593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(16253..17044,17045..17113))
                     /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     16876..16895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(17114..17218)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     17286..17304
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttcgtcttc aagaattaat tctcatgttt gacagcttat catcgataag ctgactcatg
       61 ttggtattgt gaaatagacg cagatcggga acactgaaaa ataacagtta ttattcggag
      121 ctctgtgtgg aattgtgagc ggataacaat ttcacacact aacccctact tgacagcaat
      181 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      241 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      301 caacttgaga agatcaaaaa acaactaatt attcgaaatg actgttgacg ctttgactca
      361 gccacaccac ttgttgtcct tggcctggaa cgatactcaa cagcacggtt cctggtttgc
      421 ccctcttgtc actacctccg ccggtttgtt gtgtttgttg ttgtaccttt gttcctccgg
      481 tagaagatcc gacttgcctg tcttcaatcc taagacctgg tgggagttga ccactatgag
      541 agctaagcgt gatttcgatg ctaacgcccc atcctggatt gagtcctggt tctcccagaa
      601 cgacaagcca atcagattta ttgttgactc tggttactgc accatcttgc cttcctctat
      661 ggctgacgag ttccgtaaga tgaaggagtt gtgcatgtac aagttcttgg gtactgactt
      721 ccactcccac cttccaggtt ttgatggatt taaagaggtc accagagacg cccatttgat
      781 caccaaggtt gtcatgaacc agttccagac ccaggcccca aagtatgtca agccattggc
      841 caacgaggcc tccggaatta tcaccgacat cttcggtgac tctaacgagt ggcataccgt
      901 cccagtctac aaccagtgct tggacttggt cactcgtacc gtcaccttca ttatggtcgg
      961 atctaagttg gctcacaacg aagagtggtt ggacatcgcc aagcatcacg ccgttaccat
     1021 ggctatccag gccagacagt tgcgtttgtg gccagtcatt ttgcgtccat tggttcactg
     1081 gcttgaacct caaggagcca agttgagagc tcaagttaga agagctagac aattgttgga
     1141 tccaattatt caagaacgta gagctgagcg tgatgcttgc agagctaagg gtattgagcc
     1201 acctagatac gtcgattcca ttcagtggtt cgaggacact gccaagggaa aatggtacga
     1261 cgctgctgga gctcaattgg ctatggattt cgccggaatc tacggtacct ctgacttgtt
     1321 gatcggtgga ttggttgata tcgttcgtca ccctcacttg ttggagccat tgcgtgacga
     1381 gatccgtacc gttattggtc agggtggttg gaccccagcc tctttgtaca agttgaagtt
     1441 gcttgactcc tgtcttaagg agtctcagcg tgttaagcca gtcgagtgtg ccaccatgag
     1501 atcttacgct ttgcaggacg tcaccttctc caacggtacc ttcatcccta agggagaatt
     1561 ggtcgccgtt gccgccgaca gaatgtccaa tcctgaggtc tggccagaac cagccaagta
     1621 cgacccttac cgttacatga gattgcgtga ggacccagct aaggccttct ctgcccagtt
     1681 ggagaacacc aacggtgacc atatcggttt cggttggcat cctagagctt gtcctggtcg
     1741 tttcttcgcc tctaaggaga tcaaaatgat gcttgcctat ttgttgatta gatacgactg
     1801 gaaggtcgtc ccagacgaac cattgcagta ctacagacac tccttttccg tccgtatcca
     1861 tcctaccacc aagttgatga tgcgtcgtcg tgacgaggac attagattgc caggttcttt
     1921 gcatcatcat catcatcatt gagtttgtag ccttagacat gactgttcct cagttcaagt
     1981 tgggcactta cgagaagacc ggtcttgcta gattctaatc aagaggatgt cagaatgcca
     2041 tttgcctgag agatgcaggc ttcatttttg atactttttt atttgtaacc tatatagtat
     2101 aggatttttt ttgtcatttt gtttcttctc gtacgagctt gctcctgatc agcctatctc
     2161 gcagctgatg aatatcttgt ggtaggggtt tgggaaaatc attcgagttt gatgtttttc
     2221 ttggtatttc ccactcctct tcagagtaca gaagattaag tgagaagatc ttgtgtggaa
     2281 ttgtgagcgg ataacaattt cacacactaa cccctacttg acagcaatat ataaacagaa
     2341 ggaagctgcc ctgtcttaaa cctttttttt tatcatcatt attagcttac tttcataatt
     2401 gcgactggtt ccaattgaca agcttttgat tttaacgact tttaacgaca acttgagaag
     2461 atcaaaaaac aactaattat tcgaaatggc tcaactcgac actctcgacc tggtggtcct
     2521 ggtggtgctt ttggtgggta gcgccgccta cttcaccaag ggcacctact gggccgttcc
     2581 caaggacccg tatgccgcct ccggtcccgc catgaatggt ggcgccaagg cgggcaaatc
     2641 cagggacatc attgagaaaa tggaagagac tggcaagaac tgtgtgattt tctacggctc
     2701 gcagaccggt accgccgagg attatgcgtc gcgcctggcc aaggaaggct cccagcgttt
     2761 cggcctcaag accatggtcg cagatctgga agactacgat tatgagaacc tggacaagtt
     2821 ccccgaggac aaggttgcct tcttcgtcat ggccacctat ggtgagggtg aacccaccga
     2881 caacgccgtc gagttctacc agttcatctc gggtgaggac gtcgcgttcg agagcggcgc
     2941 ctctgccgac gacaagcccc tgtcctccct caagtatgtc actttcggtc tcggtaacaa
     3001 cacctatgag cactatcagg ctatggttcg caatctggat gccgctctca ccaagctggg
     3061 tgcgcagcgc attggagatg ctggtgaagg tgatgacggc gctggcacca tggaagaaga
     3121 tttcctggcc tggaaagagc ccatgtggac tgccctgtcc gaggccatga accttcagga
     3181 gcgcgaagcc gtctatgagc cggtgttctc ggtcacggaa gatgaatccc tgtcccccga
     3241 agacgaagcc gtctacctcg gtgagccgac caagggtcac cgtgacggca cccccagtgg
     3301 cccgtattcc gctcacaacc ccttcatcgc ccccatcgtc gagtctcgtg aactgttcaa
     3361 cgtcaaggac cgtaattgtc tgcacatgga gatcagcatc gctggtagca acctttctta
     3421 ccagactggt gatcacatcg cgatttggcc cacgaacgct ggtgccgagg tggaccggtt
     3481 cctccaggtg tttggtcttg agaacaagcg tcattccgtc atcaacatca agggtatcga
     3541 tgtgaccgcc aaggttccca ttccgactcc caccacgtat gatgctgctg ttcgctacta
     3601 tatggaaatt gctgcgcccg tctcccgtca gtttgtggct accctggctg cgtttgctcc
     3661 cgatgaggag actaaggcgg aaatcgtgcg tttgggtagc gacaaggact acttccacga
     3721 gaaaatcagc aaccagtgct tcaccatcgc tcaggctctt cagagtgtca cctccaagcc
     3781 cttctcggct gtcccgttct ctctgcttat cgagggtctc aataagctcc agccccgtta
     3841 ctactccatc tcttcctcct ccatggtcca gaaggataag atcagcatta ctgccgtcgt
     3901 ggaatccact cgcttgcctg gtgccgccca ccttgtcaag ggtgtcacga ccaactatct
     3961 ccttgccctg aagcaaaagc agaatgggga tccgtctccc gaccctcacg gcttaactta
     4021 tactatcact gggccccgta acaagtacga cggaatccac gttcccgttc acgtccgcca
     4081 ctccaatttc aagctcccct ctgatccctc tcggcccatt atcatggttg gccctggtac
     4141 cggtgtggct cccttccgtg gattcatcca ggagcgtgcc gccttggccg ccaagggtga
     4201 gaacgtcggt cccaccgtgt tgttctttgg atgccgcagg cgcgatgagg actttatgta
     4261 cgcagatgaa ttcaagacct accaggaaca gcttggggac aagcttcaga tcattactgc
     4321 gttttctcgt gaaacttccc agaaggtgta tgttcagcac agactgcgtg aacactccga
     4381 tctggtgagc agcctcctga agcagaaggc taacttttac gtctgcggtg acgccgccaa
     4441 catggcgcgt gaagtcaacc ttgtgcttgg ccagatcatc gcgcaacagc ggggtctccc
     4501 ggctgaacgg gccgaggaaa tggtgaagca catgcgcagc agcggcagct accaggagga
     4561 cgtgtggtca catcatcatc atcatcattg agtttgtagc cttagacatg actgttcctc
     4621 agttcaagtt gggcacttac gagaagaccg gtcttgctag attctaatca agaggatgtc
     4681 agaatgccat ttgcctgaga gatgcaggct tcatttttga tactttttta tttgtaacct
     4741 atatagtata ggattttttt tgtcattttg tttcttctcg tacgagcttg ctcctgatca
     4801 gcctatctcg cagctgatga atatcttgtg gtaggggttt gggaaaatca ttcgagtttg
     4861 atgtttttct tggtatttcc cactcctctt cagagtacag aagattaagt gagatctaac
     4921 actttgtata gcacatcgta ccagtttatc aaaatccaac aagttttcct ctgggtactg
     4981 aaattcgatt agtaataaac gggcgcactt ggtgatctcc tcatcctgtt cactagcctt
     5041 cttttcgctt aacttcatca acgagatcaa ctcgagtctg cgagcagaga actgtttcca
     5101 aagagtttca tcgctgaaat tcagcttctt tcgaataaaa tgtgtcactc ccactttatt
     5161 actcgtggtg ttatttgttt tccacgaact agttgaatcg ccatctttac taccgtcctg
     5221 ggtgacatcc ggcgaattcg ggacaaatgt gctgttccgg tagcttgtag gaagcggcat
     5281 ccgtagggca atatacgact atagcttcta aagcgtagta caatgaaatg ttcgaaggaa
     5341 caacaaacgg atttgttttt cgtaggctca acccgttgag gtgtaactct ttagcgaaag
     5401 ggtaagattg attgttcgaa gtagggcctc aaagggaaag agaaaaaaaa aaatacacca
     5461 agagttacgt aagcatatat tttttacgta aagcatgatt gaatttcagc agtattgttt
     5521 aacaaggctg atgtcgtgtg ccaatcaaaa caaaagagat tcgcataatg ccataattgg
     5581 ggtgtgtggg cgccccctaa aacgtctttc tcatcatcat ctgcaacccc catcgaacct
     5641 cattaaatca catgacttgt gcgatcctcg gtcaactcgt tccgtgcacc cattccaccc
     5701 cgggctgacc aacgcaaggt tctccgagag tccgctaccc cagatttata tcagcaacca
     5761 gtcacctttt tccgggcacg actctatatg ccctggaaaa ccggagacga tgagcctgac
     5821 tgtaaaaggt gacagaaccc ccaactctgg ttaatctctt caacaaatac tttattttct
     5881 ttcaattcaa agaacacagt atcaagtata tcaagaatgg gtgttaagcc agttactttg
     5941 tatgacgttg ctgaatacgc tggagtttcc taccaaactg tctctagagt tgttaatcaa
     6001 gcttctcatg tctccgctaa gactagagag aaggttgagg ctgctatggc tgaattgaac
     6061 tatattccaa atagagttgc tcagcagttg gctggaaagc aatctttgtt gattggagtc
     6121 gctacttctt ctttggcttt gcatgctcca tctcagattg ttgctgctat taagtccaga
     6181 gctgaccagt tgggagcttc tgttgttgtt tctatggttg agagatctgg agttgaggct
     6241 tgcaaggctg ctgttcataa cttgttggct cagagagttt ctggattgat tattaattac
     6301 ccattggacg atcaagacgc tattgccgtt gaggccgctt gtaccaacgt cccagctttg
     6361 ttcttggacg tttccgatca aactccaatt aattctatta ttttttctca cgaggatgga
     6421 actagattgg gagttgaaca cttggttgct ttgggacatc aacagattgc tttgttggct
     6481 ggaccattgt cttccgtttc tgctagattg agattggccg gatggcacaa gtacttgacc
     6541 agaaaccaga ttcaaccaat tgctgagaga gagggagatt ggtctgctat gtctggattc
     6601 cagcagacta tgcagatgtt gaacgaagga attgtcccaa ccgctatgtt ggtcgctaat
     6661 gaccaaatgg ctttgggagc tatgagagct attactgaat ctggattgag agtcggagct
     6721 gacatttctg ttgttggata tgatgacact gaggattctt cttgctacat tccaccattg
     6781 actactatta agcaagactt cagattgttg ggacagactt ctgttgatag attgttgcag
     6841 ttgtcccaag gacaagctgt taaaggaaac caattgttgc cagtttcttt ggttaagaga
     6901 aagactactt tggctccaaa cactcagact gcttccccaa gagctttggc tgactctttg
     6961 atgcaattgg ctagacaagt ctctagattg gagtctggac aaggtggcgg cggctctgtt
     7021 aacaactcca tgaaggattt cttaggcaag aaaacggtgg atggagctga tagtctcaat
     7081 ttggccgtga atctgcaaca acagcagagt tcaaacacaa ttgccaatca atcgctttcc
     7141 tcaattggat tggaaagttt tggttacggc tctggtatca aaaacgagtt taacttccaa
     7201 gacttgatag gttcaaactc tggcagttca gatccgacat tttcagtaga cgctgacgag
     7261 gcccaaaaac tcgacatttc caacaagaac agtcgtaaga gacagaaact aggtttgctg
     7321 ccggtcagca atgcaacttc ccatttgaac ggtttcaatg gaatgtccaa tggaaagtca
     7381 cactctttct cttcaccgtc tgggactaat gacgatgaac taagtggctt gatgttcaac
     7441 tcaccaagct tcaaccccct cacagttaac gattctacca acaacagcaa ccacaatata
     7501 ggtttgtctc cgatgtcatg cttattttct acagttcaag aagcatctca aaaaaagcat
     7561 ggaaattcca gtagacactt ttcataccca tctgggccgg aggacctttg gttcaatgag
     7621 ttccaaaaac aggccctcac agccaatgga gaaaatgctg tccaacaggg agatgatgct
     7681 tctaagaaca acacagccat tcctaaggac cagtcttcga actcatcgat tttcagttca
     7741 cgttctagtg cagcttctag caactcagga gacgatattg gaaggatggg cccattctcc
     7801 aaaggaccag agattgagtt caactacgat tcttttttgg aatcgttgaa ggcagagtca
     7861 ccctcttctt caaagtacaa tctgccggaa actttgaaag agtacatgac ccttagttcg
     7921 tctcatctga atagtcaaca ctccgacact ttggcaaatg gcactaacgg taactattct
     7981 agcaccgttt ccaacaactt gagcttaagt ttgaactcct tctctttctc tgacaagttc
     8041 tcattgagtc caccaacaat cactgacgcc gaaaagtttt cattgatgag aaacttcatt
     8101 gacaacatct cgccatggtt tgacactttt gacaatacca aacagtttgg aacaaaaatt
     8161 ccagttctgg ccaaaaaatg ttcttcattg tactatgcca ttctggctat atcttctcgt
     8221 caaagagaaa ggataaagaa agagcacaat gaaaaaacat tgcaatgcta ccaatactca
     8281 ctacaacagc tcatccctac tgttcaaagc tcaaataata ttgagtacat tatcacatgt
     8341 attctcctga gtgtgttcca catcatgtct agtgaacctt caacccagag ggacatcatt
     8401 gtgtcattgg caaaatacat tcaagcatgc aacataaacg gatttacatc taatgacaaa
     8461 ctggaaaaga gtattttctg gaactatgtc aatttggatt tggctacttg tgcaatcggt
     8521 gaagagtcaa tggtcattcc ttttagctac tgggttaaag agacaactga ctacaagacc
     8581 attcaagatg tgaagccatt tttcaccaag aagactagca cgacaactga cgatgacttg
     8641 gacgatatgt atgccatcta catgctgtac attagtggta gaatcattaa cctgttgaac
     8701 tgcagagatg cgaagctcaa ttttgagccc aagtgggagt ttttgtggaa tgaactcaat
     8761 gaatgggaat tgaacaaacc cttgaccttt caaagtattg ttcagttcaa ggccaatgac
     8821 gaatcgcagg gcggatcaac ttttccaact gttctattct ccaactctcg aagctgttac
     8881 agtaaccagc tgtatcatat gagctacatc atcttagtgc agaataaacc acgattatac
     8941 aaaatcccct ttactacagt ttctgcttca atgtcatctc catcggacaa caaagctggg
     9001 atgtctgctt ccagcacacc tgcttcagac caccacgctt ctggtgatca tttgtctcca
     9061 agaagtgtag agccctctct ttcgacaacg ttgagccctc cgcctaatgc aaacggtgca
     9121 ggtaacaagt tccgctctac gctctggcat gccaagcaga tctgtgggat ttctatcaac
     9181 aacaaccaca acagcaatct agcagccaaa gtgaactcat tgcaaccatt gtggcacgct
     9241 ggaaagctaa ttagttccaa gtctgaacat acacagttgc tgaaactgtt gaacaacctt
     9301 gagtgtgcaa caggctggcc tatgaactgg aagggcaagg agttaattga ctactggaat
     9361 gttgaagaat aacatcatca tcatcatcat tgagtttgta gccttagaca tgactgttcc
     9421 tcagttcaag ttgggcactt acgagaagac cggtcttgct agattctaat caagaggatg
     9481 tcagaatgcc atttgcctga gagatgcagg cttcattttt gatacttttt tatttgtaac
     9541 ctatatagta taggattttt tttgtcattt tgtttcttct cgtacgagct tgctcctgat
     9601 cagcctatct cgcagctgat gaatatcttg tggtaggggt ttgggaaaat cattcgagtt
     9661 tgatgttttt cttggtattt cccactcctc ttcagagtac agaagattaa gtgagaacta
     9721 gtagttcgtt tgtgcaagct tatcgataag ctttaatgcg gtagtttatc acagttaaat
     9781 tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt catcctcggc
     9841 accgtcaccc tggatgctgt aggcataggc ttggttatgc cggtactgcc gggcctcttg
     9901 cgggatatcg tccattccga cagcatcgcc agtcactatg gcgtgctgct agcgctatat
     9961 gcgttgatgc aatttctatg cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc
    10021 cgcccagtcc tgctcgcttc gctacttgga gccactatcg actacgcgat catggcgacc
    10081 acacccgtcc tgtggatcta tcgaatctaa atgtaagtta aaatctctaa ataattaaat
    10141 aagtcccagt ttctccatac gaaccttaac agcattgcgg tgagcatcta gaccttcaac
    10201 agcagccaga tccatcactg cttggccaat atgtttcagt ccctcaggag ttacgtcttg
    10261 tgaagtgatg aacttctgga aggttgcagt gttaactccg ctgtattgac gggcatatcc
    10321 gtacgttggc aaagtgtggt tggtaccgga ggagtaatct ccacaactct ctggagagta
    10381 ggcaccaaca aacacagatc cagcgtgttg tacttgatca acataagaag aagcattctc
    10441 gatttgcagg atcaagtgtt caggagcgta ctgattggac atttccaaag cctgctcgta
    10501 ggttgcaacc gatagggttg tagagtgtgc aatacacttg cgtacaattt caacccttgg
    10561 caactgcaca gcttggttgt gaacagcatc ttcaattctg gcaagctcct tgtctgtcat
    10621 atcgacagcc aacagaatca cctgggaatc aataccatgt tcagcttgag acagaaggtc
    10681 tgaggcaacg aaatctggat cagcgtattt atcagcaata actagaactt cagaaggccc
    10741 agcaggcatg tcaatactac acagggctga tgtgtcattt tgaaccatca tcttggcagc
    10801 agtaacgaac tggtttcctg gaccaaatat tttgtcacac ttaggaacag tttctgttcc
    10861 gtaagccata gcagctactg cctgggcgcc tcctgctagc acgatacact tagcaccaac
    10921 cttgtgggca acgtagatga cttctggggt aagggtacca tccttcttag gtggagatgc
    10981 aaaaacaatt tctttgcaac cagcaacttt ggcaggaaca cccagcatca gggaagtgga
    11041 aggcagaatt gcggttccac caggaatata gaggccaact ttctcaatag gtcttgcaaa
    11101 acgagagcag actacaccag ggcaagtctc aacttgcaac gtctccgtta gttgagcttc
    11161 atggaatttc ctgacgttat ctatagagag atcaatggct ctcttaacgt tatctggcaa
    11221 ttgcataagt tcctctggga aaggagcttc taacacaggt gtcttcaaag cgactccatc
    11281 aaacttggca gttagttcta aaagggcttt gtcaccattt tgacgaacat tgtcgacaat
    11341 tggtttgact aattccataa tctgttccgt tttctggata ggacgacgaa gggcatcttc
    11401 aatttcttgt gaggaggcct tagaaacgtc aattttgcac aattcaatac gaccttcaga
    11461 agggacttct ttaggtttgg attcttcttt aggttgttcc ttggtgtatc ctggcttggc
    11521 atctcctttc cttctagtga cctttaggga cttcatatcc aggtttctct ccacctcgtc
    11581 caacgtcaca ccgtacttgg cacatctaac taatgcaaaa taaaataagt cagcacattc
    11641 ccaggctata tcttccttgg atttagcttc tgcaagttca tcagcttcct ccctaatttt
    11701 agcgttcaac aaaacttcgt cgtcaaataa ccgtttggta taagaacctt ctggagcatt
    11761 gctcttacga tcccacaagg tggcttccat ggctctaaga ccctttgatt ggccaaaaca
    11821 ggaagtgcgt tccaagtgac agaaaccaac acctgtttgt tcaaccacaa atttcaagca
    11881 gtctccatca caatccaatt cgatacccag caacttttga gttgctccag atgtagcacc
    11941 tttataccac aaaccgtgac gacgagattg gtagactcca gtttgtgtcc ttatagcctc
    12001 cggaatagac tttttggacg agtacaccag gcccaacgag taattagaag agtcagccac
    12061 caaagtagtg aatagaccat cggggcggtc agtagtcaaa gacgccaaca aaatttcact
    12121 gacagggaac tttttgacat cttcagaaag ttcgtattca gtagtcaatt gccgagcatc
    12181 aataatgggg attataccag aagcaacagt ggaagtcaca tctaccaact ttgcggtctc
    12241 agaaaaagca taaacagttc tactaccgcc attagtgaaa cttttcaaat cgcccagtgg
    12301 agaagaaaaa ggcacagcga tactagcatt agcgggcaag gatgcaactt tatcaaccag
    12361 ggtcctatag ataaccctag cgcctgggat catcctttgg acaactcttt ctgccaaatc
    12421 taggtccaaa atcacttcat tgataccatt attgtacaac ttgagcaagt tgtcgatcag
    12481 ctcctcaaat tggtcctctg taacggatga ctcaacttgc acattaactt gaagctcagt
    12541 cgattgagtg aacttgatca ggttgtgcag ctggtcagca gcatagggaa acacggcttt
    12601 tcctaccaaa ctcaaggaat tatcaaactc tgcaacactt gcgtatgcag gtagcaaggg
    12661 aaatgtcata cttgaagtcg gacagtgagt gtagtcttga gaaattctga agccgtattt
    12721 ttattatcag tgagtcagtc atcaggagat cctctacgcc ggacgcatcg tggccgacct
    12781 gcaggggggg gggggcgctg aggtctgcct cgtgaagaag gtgttgctga ctcataccag
    12841 gcctgaatcg ccccatcatc cagccagaaa gtgagggagc cacggttgat gagagctttg
    12901 ttgtaggtgg accagttggt gattttgaac ttttgctttg ccacggaacg gtctgcgttg
    12961 tcgggaagat gcgtgatctg atccttcaac tcagcaaaag ttcgatttat tcaacaaagc
    13021 cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg
    13081 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
    13141 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
    13201 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
    13261 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
    13321 ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc
    13381 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
    13441 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
    13501 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
    13561 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
    13621 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
    13681 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
    13741 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
    13801 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
    13861 gcctcgagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
    13921 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
    13981 agagattttg agacacaacg tggctttccc ccccccccct gcaggtcggc atcaccggcg
    14041 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
    14101 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
    14161 ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg
    14221 gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag
    14281 tatctatgat tggaagtatg ggaatggtga tacccgcatt cttcagtgtc ttgaggtctc
    14341 ctatcagatt atgcccaact aaagcaaccg gaggaggaga tttcatggta aatttctctg
    14401 acttttggtc atcagtagac tcgaactgtg agactatctc ggttatgaca gcagaaatgt
    14461 ccttcttgga gacagtaaat gaagtcccac caataaagaa atccttgtta tcaggaacaa
    14521 acttcttgtt tcgaactttt tcggtgcctt gaactataaa atgtagagtg gatatgtcgg
    14581 gtaggaatgg agcgggcaaa tgcttacctt ctggaccttc aagaggtatg tagggtttgt
    14641 agatactgat gccaacttca gtgacaacgt tgctatttcg ttcaaaccat tccgaatcca
    14701 gagaaatcaa agttgtttgt ctactattga tccaagccag tgcggtcttg aaactgacaa
    14761 tagtgtgctc gtgttttgag gtcatctttg tatgaataaa tctagtcttt gatctaaata
    14821 atcttgacga gccagacgat aataccaatc taaactcttt aaacgttaaa ggacaagtat
    14881 gtctgcctgt attaaacccc aaatcagctc gtagtctgat cctcatcaac ttgaggggca
    14941 ctatcttgtt ttagagaaat ttgcggagat gcgatatcga gaaaaaggta cgctgatttt
    15001 aaacgtgaaa tttatctcaa gatctgctgc ctcgcgcgtt tcggtgatga cggtgaaaac
    15061 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
    15121 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
    15181 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
    15241 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
    15301 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
    15361 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
    15421 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    15481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    15541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    15601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    15661 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    15721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    15781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    15841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    15901 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
    15961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    16021 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    16081 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    16141 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    16201 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    16261 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    16321 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    16381 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    16441 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    16501 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    16561 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    16621 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    16681 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    16741 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    16801 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    16861 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    16921 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    16981 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    17041 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    17101 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    17161 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    17221 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
    17281 ataaaaatag gcgtatcacg aggccc
//
