LOCUS       sequence-246575-       12504 bp    DNA     circular SYN 15-APR-2025
DEFINITION  yeast plasmid overexpressing CPR and sLovA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12504)
  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 12504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_lacO-cP-AOX1-sAR
FEATURES             Location/Qualifiers
     source          1..12504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    256..272
                     /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)."
     terminator      complement(288..534)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(614..631)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2404..2420
                     /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)."
     primer_bind     complement(2555..2573)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2641..2745
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2746..2814,2815..3606)
                     /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     complement(2964..2983)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3777..4365
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4266..4285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4519..4536
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4551..4691
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4617..4636)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4777..4799
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4835..5583
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     primer_bind     5778..5797
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5951..5970)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5962..6777
                     /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             5962..6777
                     /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     complement(6035..6054)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             7190..9724
                     /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"
     terminator      complement(10143..10389)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(10469..10486)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 agtaggcggc gctacccacc aaaagcacca ccaggaccac caggtcgaga gtgtcgagtt
       61 gagccatttc gaataattag ttgttttttg atcttctcaa gttgtcgtta aaagtcgtta
      121 aaatcaaaag cttgtcaatt ggaaccagtc gcaattatga aagtaagcta ataatgatga
      181 taaaaaaaaa ggtttaagac agggcagctt ccttctgttt atatattgct gtcaagtagg
      241 ggttagtgtg tgaaattgtt atccgctcac aattccacac aagatcttct cacttaatct
      301 tctgtactct gaagaggagt gggaaatacc aagaaaaaca tcaaactcga atgattttcc
      361 caaaccccta ccacaagata ttcatcagct gcgagatagg ctgatcagga gcaagctcgt
      421 acgagaagaa acaaaatgac aaaaaaaatc ctatactata taggttacaa ataaaaaagt
      481 atcaaaaatg aagcctgcat ctctcaggca aatggcattc tgacatcctc ttgattagaa
      541 tctagcaaga ccggtcttct cgtaagtgcc caacttgaac tgaggaacag tcatgtctaa
      601 ggctacaaac tcaatgatga tgatgatgat gcaaagaacc tggcaatcta atgtcctcgt
      661 cacgacgacg catcatcaac ttggtggtag gatggatacg gacggaaaag gagtgtctgt
      721 agtactgcaa tggttcgtct gggacgacct tccagtcgta tctaatcaac aaataggcaa
      781 gcatcatttt gatctcctta gaggcgaaga aacgaccagg acaagctcta ggatgccaac
      841 cgaaaccgat atggtcaccg ttggtgttct ccaactgggc agagaaggcc ttagctgggt
      901 cctcacgcaa tctcatgtaa cggtaagggt cgtacttggc tggttctggc cagacctcag
      961 gattggacat tctgtcggcg gcaacggcga ccaattctcc cttagggatg aaggtaccgt
     1021 tggagaaggt gacgtcctgc aaagcgtaag atctcatggt ggcacactcg actggcttaa
     1081 cacgctgaga ctccttaaga caggagtcaa gcaacttcaa cttgtacaaa gaggctgggg
     1141 tccaaccacc ctgaccaata acggtacgga tctcgtcacg caatggctcc aacaagtgag
     1201 ggtgacgaac gatatcaacc aatccaccga tcaacaagtc agaggtaccg tagattccgg
     1261 cgaaatccat agccaattga gctccagcag cgtcgtacca ttttcccttg gcagtgtcct
     1321 cgaaccactg aatggaatcg acgtatctag gtggctcaat acccttagct ctgcaagcat
     1381 cacgctcagc tctacgttct tgaataattg gatccaacaa ttgtctagct cttctaactt
     1441 gagctctcaa cttggctcct tgaggttcaa gccagtgaac caatggacgc aaaatgactg
     1501 gccacaaacg caactgtctg gcctggatag ccatggtaac ggcgtgatgc ttggcgatgt
     1561 ccaaccactc ttcgttgtga gccaacttag atccgaccat aatgaaggtg acggtacgag
     1621 tgaccaagtc caagcactgg ttgtagactg ggacggtatg ccactcgtta gagtcaccga
     1681 agatgtcggt gataattccg gaggcctcgt tggccaatgg cttgacatac tttggggcct
     1741 gggtctggaa ctggttcatg acaaccttgg tgatcaaatg ggcgtctctg gtgacctctt
     1801 taaatccatc aaaacctgga aggtgggagt ggaagtcagt acccaagaac ttgtacatgc
     1861 acaactcctt catcttacgg aactcgtcag ccatagagga aggcaagatg gtgcagtaac
     1921 cagagtcaac aataaatctg attggcttgt cgttctggga gaaccaggac tcaatccagg
     1981 atggggcgtt agcatcgaaa tcacgcttag ctctcatagt ggtcaactcc caccaggtct
     2041 taggattgaa gacaggcaag tcggatcttc taccggagga acaaaggtac aacaacaaac
     2101 acaacaaacc ggcggaggta gtgacaagag gggcaaacca ggaaccgtgc tgttgagtat
     2161 cgttccaggc caaggacaac aagtggtgtg gctgagtcaa agcgtcaaca gtcatttcga
     2221 ataattagtt gttttttgat cttctcaagt tgtcgttaaa agtcgttaaa atcaaaagct
     2281 tgtcaattgg aaccagtcgc aattatgaaa gtaagctaat aatgatgata aaaaaaaagg
     2341 tttaagacag ggcagcttcc ttctgtttat atattgctgt caagtagggg ttagtgtgtg
     2401 aaattgttat ccgctcacaa ttccacacag agctccgaat aataactgtt atttttcagt
     2461 gttcccgatc tgcgtctatt tcacaatacc aacatgagtc agcttatcga tgataagctg
     2521 tcaaacatga gaattaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     2581 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2641 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2701 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2761 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2821 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2881 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2941 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     3001 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3061 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3121 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3181 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3241 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     3301 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3361 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3421 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3481 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3541 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3601 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3661 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3721 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3781 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3841 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3901 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3961 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4021 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4081 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4141 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4201 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4261 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4321 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4381 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4441 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4501 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     4561 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     4621 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     4681 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4741 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4801 tttcaccgtc atcaccgaaa cgcgcgaggc agcagatctt gagataaatt tcacgtttaa
     4861 aatcagcgta cctttttctc gatatcgcat ctccgcaaat ttctctaaaa caagatagtg
     4921 cccctcaagt tgatgaggat cagactacga gctgatttgg ggtttaatac aggcagacat
     4981 acttgtcctt taacgtttaa agagtttaga ttggtattat cgtctggctc gtcaagatta
     5041 tttagatcaa agactagatt tattcataca aagatgacct caaaacacga gcacactatt
     5101 gtcagtttca agaccgcact ggcttggatc aatagtagac aaacaacttt gatttctctg
     5161 gattcggaat ggtttgaacg aaatagcaac gttgtcactg aagttggcat cagtatctac
     5221 aaaccctaca tacctcttga aggtccagaa ggtaagcatt tgcccgctcc attcctaccc
     5281 gacatatcca ctctacattt tatagttcaa ggcaccgaaa aagttcgaaa caagaagttt
     5341 gttcctgata acaaggattt ctttattggt gggacttcat ttactgtctc caagaaggac
     5401 atttctgctg tcataaccga gatagtctca cagttcgagt ctactgatga ccaaaagtca
     5461 gagaaattta ccatgaaatc tcctcctccg gttgctttag ttgggcataa tctgatagga
     5521 gacctcaaga cactgaagaa tgcgggtatc accattccca tacttccaat catagatact
     5581 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     5641 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     5701 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     5761 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     5821 ccggtgatgc cgacctgcag gggggggggg gaaagccacg ttgtgtctca aaatctctga
     5881 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     5941 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg ctcgaggccg
     6001 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     6061 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
     6121 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     6181 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     6241 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
     6301 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     6361 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     6421 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     6481 cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc
     6541 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     6601 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     6661 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     6721 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     6781 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcggc
     6841 tttgttgaat aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca
     6901 acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca
     6961 aagctctcat caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct
     7021 ggtatgagtc agcaacacct tcttcacgag gcagacctca gcgccccccc ccccctgcag
     7081 gtcggccacg atgcgtccgg cgtagaggat ctcctgatga ctgactcact gataataaaa
     7141 atacggcttc agaatttctc aagactacac tcactgtccg acttcaagta tgacatttcc
     7201 cttgctacct gcatacgcaa gtgttgcaga gtttgataat tccttgagtt tggtaggaaa
     7261 agccgtgttt ccctatgctg ctgaccagct gcacaacctg atcaagttca ctcaatcgac
     7321 tgagcttcaa gttaatgtgc aagttgagtc atccgttaca gaggaccaat ttgaggagct
     7381 gatcgacaac ttgctcaagt tgtacaataa tggtatcaat gaagtgattt tggacctaga
     7441 tttggcagaa agagttgtcc aaaggatgat cccaggcgct agggttatct ataggaccct
     7501 ggttgataaa gttgcatcct tgcccgctaa tgctagtatc gctgtgcctt tttcttctcc
     7561 actgggcgat ttgaaaagtt tcactaatgg cggtagtaga actgtttatg ctttttctga
     7621 gaccgcaaag ttggtagatg tgacttccac tgttgcttct ggtataatcc ccattattga
     7681 tgctcggcaa ttgactactg aatacgaact ttctgaagat gtcaaaaagt tccctgtcag
     7741 tgaaattttg ttggcgtctt tgactactga ccgccccgat ggtctattca ctactttggt
     7801 ggctgactct tctaattact cgttgggcct ggtgtactcg tccaaaaagt ctattccgga
     7861 ggctataagg acacaaactg gagtctacca atctcgtcgt cacggtttgt ggtataaagg
     7921 tgctacatct ggagcaactc aaaagttgct gggtatcgaa ttggattgtg atggagactg
     7981 cttgaaattt gtggttgaac aaacaggtgt tggtttctgt cacttggaac gcacttcctg
     8041 ttttggccaa tcaaagggtc ttagagccat ggaagccacc ttgtgggatc gtaagagcaa
     8101 tgctccagaa ggttcttata ccaaacggtt atttgacgac gaagttttgt tgaacgctaa
     8161 aattagggag gaagctgatg aacttgcaga agctaaatcc aaggaagata tagcctggga
     8221 atgtgctgac ttattttatt ttgcattagt tagatgtgcc aagtacggtg tgacgttgga
     8281 cgaggtggag agaaacctgg atatgaagtc cctaaaggtc actagaagga aaggagatgc
     8341 caagccagga tacaccaagg aacaacctaa agaagaatcc aaacctaaag aagtcccttc
     8401 tgaaggtcgt attgaattgt gcaaaattga cgtttctaag gcctcctcac aagaaattga
     8461 agatgccctt cgtcgtccta tccagaaaac ggaacagatt atggaattag tcaaaccaat
     8521 tgtcgacaat gttcgtcaaa atggtgacaa agccctttta gaactaactg ccaagtttga
     8581 tggagtcgct ttgaagacac ctgtgttaga agctcctttc ccagaggaac ttatgcaatt
     8641 gccagataac gttaagagag ccattgatct ctctatagat aacgtcagga aattccatga
     8701 agctcaacta acggagacgt tgcaagttga gacttgccct ggtgtagtct gctctcgttt
     8761 tgcaagacct attgagaaag ttggcctcta tattcctggt ggaaccgcaa ttctgccttc
     8821 cacttccctg atgctgggtg ttcctgccaa agttgctggt tgcaaagaaa ttgtttttgc
     8881 atctccacct aagaaggatg gtacccttac cccagaagtc atctacgttg cccacaaggt
     8941 tggtgctaag tgtatcgtgc tagcaggagg cgcccaggca gtagctgcta tggcttacgg
     9001 aacagaaact gttcctaagt gtgacaaaat atttggtcca ggaaaccagt tcgttactgc
     9061 tgccaagatg atggttcaaa atgacacatc agccctgtgt agtattgaca tgcctgctgg
     9121 gccttctgaa gttctagtta ttgctgataa atacgctgat ccagatttcg ttgcctcaga
     9181 ccttctgtct caagctgaac atggtattga ttcccaggtg attctgttgg ctgtcgatat
     9241 gacagacaag gagcttgcca gaattgaaga tgctgttcac aaccaagctg tgcagttgcc
     9301 aagggttgaa attgtacgca agtgtattgc acactctaca accctatcgg ttgcaaccta
     9361 cgagcaggct ttggaaatgt ccaatcagta cgctcctgaa cacttgatcc tgcaaatcga
     9421 gaatgcttct tcttatgttg atcaagtaca acacgctgga tctgtgtttg ttggtgccta
     9481 ctctccagag agttgtggag attactcctc cggtaccaac cacactttgc caacgtacgg
     9541 atatgcccgt caatacagcg gagttaacac tgcaaccttc cagaagttca tcacttcaca
     9601 agacgtaact cctgagggac tgaaacatat tggccaagca gtgatggatc tggctgctgt
     9661 tgaaggtcta gatgctcacc gcaatgctgt taaggttcgt atggagaaac tgggacttat
     9721 ttaattattt agagatttta acttacattt agattcgata gatccacagg acgggtgtgg
     9781 tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg actgggcggc
     9841 ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc atcaacgcat
     9901 atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg atatcccgca
     9961 agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc
    10021 cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat
    10081 ttaactgtga taaactaccg cattaaagct tatcgataag cttgcacaaa cgaactacta
    10141 gttctcactt aatcttctgt actctgaaga ggagtgggaa ataccaagaa aaacatcaaa
    10201 ctcgaatgat tttcccaaac ccctaccaca agatattcat cagctgcgag ataggctgat
    10261 caggagcaag ctcgtacgag aagaaacaaa atgacaaaaa aaatcctata ctatataggt
    10321 tacaaataaa aaagtatcaa aaatgaagcc tgcatctctc aggcaaatgg cattctgaca
    10381 tcctcttgat tagaatctag caagaccggt cttctcgtaa gtgcccaact tgaactgagg
    10441 aacagtcatg tctaaggcta caaactcaat gatgatgatg atgatgtgac cacacgtcct
    10501 cctggtagct gccgctgctg cgcatgtgct tcaccatttc ctcggcccgt tcagccggga
    10561 gaccccgctg ttgcgcgatg atctggccaa gcacaaggtt gacttcacgc gccatgttgg
    10621 cggcgtcacc gcagacgtaa aagttagcct tctgcttcag gaggctgctc accagatcgg
    10681 agtgttcacg cagtctgtgc tgaacataca ccttctggga agtttcacga gaaaacgcag
    10741 taatgatctg aagcttgtcc ccaagctgtt cctggtaggt cttgaattca tctgcgtaca
    10801 taaagtcctc atcgcgcctg cggcatccaa agaacaacac ggtgggaccg acgttctcac
    10861 ccttggcggc caaggcggca cgctcctgga tgaatccacg gaagggagcc acaccggtac
    10921 cagggccaac catgataatg ggccgagagg gatcagaggg gagcttgaaa ttggagtggc
    10981 ggacgtgaac gggaacgtgg attccgtcgt acttgttacg gggcccagtg atagtataag
    11041 ttaagccgtg agggtcggga gacggatccc cattctgctt ttgcttcagg gcaaggagat
    11101 agttggtcgt gacacccttg acaaggtggg cggcaccagg caagcgagtg gattccacga
    11161 cggcagtaat gctgatctta tccttctgga ccatggagga ggaagagatg gagtagtaac
    11221 ggggctggag cttattgaga ccctcgataa gcagagagaa cgggacagcc gagaagggct
    11281 tggaggtgac actctgaaga gcctgagcga tggtgaagca ctggttgctg attttctcgt
    11341 ggaagtagtc cttgtcgcta cccaaacgca cgatttccgc cttagtctcc tcatcgggag
    11401 caaacgcagc cagggtagcc acaaactgac gggagacggg cgcagcaatt tccatatagt
    11461 agcgaacagc agcatcatac gtggtgggag tcggaatggg aaccttggcg gtcacatcga
    11521 tacccttgat gttgatgacg gaatgacgct tgttctcaag accaaacacc tggaggaacc
    11581 ggtccacctc ggcaccagcg ttcgtgggcc aaatcgcgat gtgatcacca gtctggtaag
    11641 aaaggttgct accagcgatg ctgatctcca tgtgcagaca attacggtcc ttgacgttga
    11701 acagttcacg agactcgacg atgggggcga tgaaggggtt gtgagcggaa tacgggccac
    11761 tgggggtgcc gtcacggtga cccttggtcg gctcaccgag gtagacggct tcgtcttcgg
    11821 gggacaggga ttcatcttcc gtgaccgaga acaccggctc atagacggct tcgcgctcct
    11881 gaaggttcat ggcctcggac agggcagtcc acatgggctc tttccaggcc aggaaatctt
    11941 cttccatggt gccagcgccg tcatcacctt caccagcatc tccaatgcgc tgcgcaccca
    12001 gcttggtgag agcggcatcc agattgcgaa ccatagcctg atagtgctca taggtgttgt
    12061 taccgagacc gaaagtgaca tacttgaggg aggacagggg cttgtcgtcg gcagaggcgc
    12121 cgctctcgaa cgcgacgtcc tcacccgaga tgaactggta gaactcgacg gcgttgtcgg
    12181 tgggttcacc ctcaccatag gtggccatga cgaagaaggc aaccttgtcc tcggggaact
    12241 tgtccaggtt ctcataatcg tagtcttcca gatctgcgac catggtcttg aggccgaaac
    12301 gctgggagcc ttccttggcc aggcgcgacg cataatcctc ggcggtaccg gtctgcgagc
    12361 cgtagaaaat cacacagttc ttgccagtct cttccatttt ctcaatgatg tccctggatt
    12421 tgcccgcctt ggcgccacca ttcatggcgg gaccggaggc ggcatacggg tccttgggaa
    12481 cggcccagta ggtgcccttg gtga
//
