LOCUS       sequence-396442-       12110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli/Yeast shuttle vector carrying CloNAT marker and Spcas9 
            allowing cloning of gRNA flanked by tRNA promoter and HDV ribozyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12110)
  AUTHORS   Kuanyshev N, Rao CV, Dien B, Jin YS
  TITLE     Domesticating a food spoilage yeast into an organic acid-tolerant 
            metabolic engineering host: Lactic acid production by engineered 
            Zygosaccharomyces bailii.
  JOURNAL   Biotechnol Bioeng. 2021 Jan;118(1):372-382. doi: 10.1002/bit.27576. 
            Epub 2020 Oct 8.
  PUBMED    33030791
REFERENCE   2  (bases 1 to 12110)
  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"; doi: "10.1002/bit.27576";
            journalName: "Biotechnol Bioeng"; date: "2021-01"; volume: "118"; 
            issue: "1"; pages: "372-382"
COMMENT     Sequence Label: pCast
FEATURES             Location/Qualifiers
     source          1..12110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            2061..3180
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        2061..2404
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2405..2977
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2983..3180
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             3802..3825
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3829..3849
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3850..7950
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTYKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSITHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7954..7974
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        8893..9559
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     9429..9452
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     misc_RNA        9664..9739
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           9744..9811
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     primer_bind     complement(9853..9871)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(join(10305..11096,11097..11165))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGTRVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10928..10947
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11166..11270)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      11373..11961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11862..11881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcatcaatag gcacgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca
       61 gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat
      121 catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc
      181 tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg
      241 cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac
      301 ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaggat
      361 ccatgccaca atcttgggaa gaattggctg ctgacaagag agctagattg gctaagacta
      421 tcccagacga atggaaggtt caaactttgc cagctgaaga ctctgttatc gacttcccaa
      481 agaagtctgg tatcttgtct gaagctgaat tgaagatcac tgaagcttct gctgctgact
      541 tggtttctaa gttggctgct ggtgaattga cttctgttga agttactttg gctttctgta
      601 agagagctgc tatcgctcaa caattgacta actgtgctca cgaattcttc ccagacgctg
      661 ctttggctca agctagagaa ttggacgaat actacgctaa gcacaaaaga cctgttggtc
      721 cattgcacgg tttgccaatc tctttgaagg accaattgag agttaagggt tacgaaactt
      781 ctatgggtta catctcttgg ttgaacaagt acgacgaagg tgactctgtt ttgactacta
      841 tgttgagaaa ggctggtgct gttttctacg ttaagacttc tgttccacaa actttgatgg
      901 tttgtgaaac tgttaacaac atcatcggta gaactgttaa cccaagaaac aagaactggt
      961 cttgtggtgg ttcttctggt ggtgaaggtg ctatcgttgg tatcagaggt ggtgttatcg
     1021 gtgttggtac tgacatcggt ggttctatca gagttccagc tgctttcaac ttcttgtacg
     1081 gtttaagacc atctcacggt agattgccat acgctaagat ggctaactct atggaaggtc
     1141 aagaaactgt acactctgtt gttggtccaa tcactcactc tgttgaagac ttgagattgt
     1201 tcactaagtc tgttttgggt caagaaccat ggaagtacga ctctaaggtt atcccaatgc
     1261 catggagaca atctgaatct gacatcatcg cttctaagat caagaacggt ggtttgaaca
     1321 tcggttacta caacttcgac ggtaacgttt tgccacaccc accaatcttg agaggtgttg
     1381 aaactactgt tgctgctttg gctaaggctg gtcacactgt tactccatgg actccataca
     1441 agcacgactt cggtcacgac ttgatctctc acatctacgc tgctgacggt tctgctgacg
     1501 ttatgagaga catctctgct tctggtgaac cagctatccc aaacatcaag gacttgttga
     1561 acccaaacat caaggctgtt aacatgaacg aattgtggga cactcacttg caaaagtgga
     1621 actaccaaat ggaatacttg gaaaagtgga gagaagctga agaaaaggct ggtaaggaat
     1681 tggacgctat catcgctcca atcactccaa ctgctgctgt tagacacgac caattcagat
     1741 actacggtta cgcttctgtt atcaacttgt tggacttcac ttctgttgtt gttccagtta
     1801 ctttcgctga caagaacatc gacaagaaga acgaatcttt caaggctgtt tctgaattgg
     1861 acgctttggt tcaagaagaa tacgacccag aagcttacca cggtgctcca gttgctgttc
     1921 aagttatcgg tagaagattg tctgaagaaa gaactttggc tatcgctgaa gaagttggta
     1981 agttgttggg taacgttgtt actccataac tcgagggatc tgtttagctt gccttgtccc
     2041 cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg
     2101 tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg
     2161 tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct
     2221 cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc
     2281 acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc
     2341 atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac
     2401 aaccatgggt accactcttg acgacacggc ttaccggtac cgcaccagtg tcccggggga
     2461 cgccgaggcc atcgaggcac tggatgggtc cttcaccacc gacaccgtct tccgcgtcac
     2521 cgccaccggg gacggcttca ccctgcggga ggtgccggtg gacccgcccc tgaccaaggt
     2581 gttccccgac gacgaatcgg acgacgaatc ggacgacggg gaggacggcg acccggactc
     2641 ccggacgttc gtcgcgtacg gggacgacgg cgacctggcg ggcttcgtgg tcgtctcgta
     2701 ctccggctgg aaccgccggc tgaccgtcga ggacatcgag gtcgccccgg agcaccgggg
     2761 gcacggggtc gggcgcgcgt tgatggggct cgcgacggag ttcgcccgcg agcggggcgc
     2821 cgggcacctc tggctggagg tcaccaacgt caacgcaccg gcgatccacg cgtaccggcg
     2881 gatggggttc accctctgcg gcctggacac cgccctgtac gacggcaccg cctcggacgg
     2941 cgagcaggcg ctctacatga gcatgccctg cccctaatca gtactgacaa taaaaagatt
     3001 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     3061 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     3121 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     3181 ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa cgcctcgagg acaataaaaa
     3241 gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt
     3301 ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc
     3361 gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat
     3421 actggagctc tcgagaaccc tcacctttcg agaggacgat gcccgtgtct aaatgattcg
     3481 accagcctaa gaatgttcaa caatctataa tcagtccata gtcaacaaga gccctcaaag
     3541 ttgatatcgt ttagtgatct ccctcgagat gagcggagga tggttgcttt ttggggagcc
     3601 ctaagtggta cgtgcggcga gcgggatgtt tccccattag gcaaaggacg accgggtaac
     3661 ccctcgagaa aaaaaatttt tcgccgctaa tctggtgtta tataaagctc cccctgctcc
     3721 tggaattttt ttccttgtca actcacaccg gaaatcaagg catttcattc tgagtagttc
     3781 ttaaaaacat aatcaacaat ggattataaa gatgacgatg acaaacctcc aaaaaagaag
     3841 agaaaggtcg ataagaaata ctcaataggc ttagatatcg gcacaaatag cgtcggatgg
     3901 gcggtgatca ctgatgaata taaggttccg tctaaaaagt tcaaggttct gggaaataca
     3961 gaccgccaca gtatcaaaaa aaatcttata ggggctcttt tatttgacag tggagagaca
     4021 gcggaagcga ctcgtctcaa acggacagct cgtagaaggt atacacgtcg gaagaatcgt
     4081 atttgttatc tacaggagat tttttcaaat gagatggcga aagtagatga tagtttcttt
     4141 catcgacttg aagagtcttt tttggtggaa gaagacaaga agcatgaacg tcatcctatt
     4201 tttggaaata tagtagatga agttgcttat catgagaaat atccaactat ctatcatctg
     4261 cgaaaaaaat tggtagattc tacttataaa gcggatttgc gcttaatcta tttggcctta
     4321 gcgcatatga ttaagtttcg tggtcatttt ttgattgagg gagatttaaa tcctgataat
     4381 agtgatgtgg acaaactatt tatccagttg gtacaaacct acaatcaatt atttgaagaa
     4441 aaccctatta acgcaagtgg agtagatgct aaagcgattc tttctgcacg attgagtaaa
     4501 tcaagacgat tagaaaatct cattgctcag ctccccggtg agaagaaaaa tggcttattt
     4561 gggaatctca ttgctttgtc attgggtttg acccctaatt ttaaatcaaa ttttgatttg
     4621 gcagaagatg ctaaattaca gctttcaaaa gatacttacg atgatgattt agataattta
     4681 ttggcgcaaa ttggagatca atatgctgat ttgtttttgg cagctaagaa tttatcagat
     4741 gctattttac tttcagatat cctaagagta aatactgaaa taactaaggc tcccctatca
     4801 gcttcaatga ttaaacgcta cgatgaacat catcaagact tgactctttt aaaagcttta
     4861 gttcgacaac aacttccaga aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga
     4921 tatgcaggtt atattgatgg gggagctagc caagaagaat tttataaatt tatcaaacca
     4981 attttagaaa aaatggatgg tactgaggaa ttattggtga aactaaatcg tgaagatttg
     5041 ctgcgcaagc aacggacctt tgacaacggc tctattaccc atcaaattca cttgggtgag
     5101 ctgcatgcta ttttgagaag acaagaagac ttttatccat ttttaaaaga caatcgtgag
     5161 aagattgaaa aaatcttgac ttttcgaatt ccttattatg ttggtccatt ggcgcgtggc
     5221 aatagtcgtt ttgcatggat gactcggaag tctgaagaaa caattacccc atggaatttt
     5281 gaagaagttg tcgataaagg tgcttcagct caatcattta ttgaacgcat gacaaacttt
     5341 gataaaaatc ttccaaatga aaaagtacta ccaaaacata gtttgcttta tgagtatttt
     5401 acggtttata acgaattgac aaaggtcaaa tatgttactg aaggaatgcg aaaaccagca
     5461 tttctttcag gtgaacagaa gaaagccatt gttgatttac tcttcaaaac aaatcgaaaa
     5521 gtaaccgtta agcaattaaa agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt
     5581 gaaatttcag gagttgaaga tagatttaat gcttcattag gtacctacca tgatttgcta
     5641 aaaattatta aagataaaga ttttttggat aatgaagaaa atgaagatat cttagaggat
     5701 attgttttaa cattgacctt atttgaagat agggagatga ttgaggaaag acttaaaaca
     5761 tatgctcacc tctttgatga taaggtgatg aaacagctta aacgtcgccg ttatactggt
     5821 tggggacgtt tgtctcgaaa attgattaat ggtattaggg ataagcaatc tggcaaaaca
     5881 atattagatt ttttgaaatc agatggtttt gccaatcgca attttatgca gctgatccat
     5941 gatgatagtt tgacatttaa agaagacatt caaaaagcac aagtgtctgg acaaggcgat
     6001 agtttacatg aacatattgc aaatttagct ggtagccctg ctattaaaaa aggtatttta
     6061 cagactgtaa aagttgttga tgaattggtc aaagtaatgg ggcggcataa gccagaaaat
     6121 atcgttattg aaatggcacg tgaaaatcag acaactcaaa agggccagaa aaattcgcga
     6181 gagcgtatga aacgaatcga agaaggtatc aaagaattag gaagtcagat tcttaaagag
     6241 catcctgttg aaaatactca attgcaaaat gaaaagctct atctctatta tctccaaaat
     6301 ggaagagaca tgtatgtgga ccaagaatta gatattaatc gtttaagtga ttatgatgtc
     6361 gatcacattg ttccacaaag tttccttaaa gacgattcaa tagacaataa ggtcttaacg
     6421 cgttctgata aaaatcgtgg taaatcggat aacgttccaa gtgaagaagt agtcaaaaag
     6481 atgaaaaact attggagaca acttctaaac gccaagttaa tcactcaacg taagtttgat
     6541 aatttaacga aagctgaacg tggaggtttg agtgaacttg ataaagctgg ttttatcaaa
     6601 cgccaattgg ttgaaactcg ccaaatcact aagcatgtgg cacaaatttt ggatagtcgc
     6661 atgaatacta aatacgatga aaatgataaa cttattcgag aggttaaagt gattacctta
     6721 aaatctaaat tagtttctga cttccgaaaa gatttccaat tctataaagt acgtgagatt
     6781 aacaattacc atcatgccca tgatgcgtat ctaaatgccg tcgttggaac tgctttgatt
     6841 aagaaatatc caaaacttga atcggagttt gtctatggtg attataaagt ttatgatgtt
     6901 cgtaaaatga ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa atatttcttt
     6961 tactctaata tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg agagattcgc
     7021 aaacgccctc taatcgaaac taatggggaa actggagaaa ttgtctggga taaagggcga
     7081 gattttgcca cagtgcgcaa agtattgtcc atgccccaag tcaatattgt caagaaaaca
     7141 gaagtacaga caggcggatt ctccaaggag tcaattttac caaaaagaaa ttcggacaag
     7201 cttattgctc gtaaaaaaga ctgggatcca aaaaaatatg gtggttttga tagtccaacg
     7261 gtagcttatt cagtcctagt ggttgctaag gtggaaaaag ggaaatcgaa gaagttaaaa
     7321 tccgttaaag agttactagg gatcacaatt atggaaagaa gttcctttga aaaaaatccg
     7381 attgactttt tagaagctaa aggatataag gaagttaaaa aagacttaat cattaaacta
     7441 cctaaatata gtctttttga gttagaaaac ggtcgtaaac ggatgctggc tagtgccgga
     7501 gaattacaaa aaggaaatga gctggctctg ccaagcaaat atgtgaattt tttatattta
     7561 gctagtcatt atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt
     7621 gtggagcagc ataagcatta tttagatgag attattgagc aaatcagtga attttctaag
     7681 cgtgttattt tagcagatgc caatttagat aaagttctta gtgcatataa caaacataga
     7741 gacaaaccaa tacgtgaaca agcagaaaat attattcatt tatttacgtt gacgaatctt
     7801 ggagctcccg ctgcttttaa atattttgat acaacaattg atcgtaaacg atatacgtct
     7861 acaaaagaag ttttagatgc cactcttatc catcaatcca tcactggtct ttatgaaaca
     7921 cgcattgatt tgagtcagct aggaggtgac cctccaaaaa agaagagaaa ggtctgagcg
     7981 gttttgtata actaaataat attggaaact aaatacgaat acccaaattt tttatctaaa
     8041 ttttgccgaa agattaaaat ctgcagagat atccgaaaca ggtaaatgga tgtttcaatc
     8101 cctgtagtca gtcaggaacc catattatat tacagtatta gtcgccgctt aggcacgcct
     8161 ttaattagca aaatcaaacc ttaagtgcat atgccgtata agggaaactc aaagaactgg
     8221 catcgcaaaa atgaaaacat acgttgaaac tacggcaaag gattggtcag atcgcttcat
     8281 acagggaaag ttcggcatca acatctttgg ataatatcag aatgagaaag aacagatacg
     8341 cagtacgttt tttggtgagc tctttgcact tctttagttc tttccatcaa tatcagtttt
     8401 ttaaactttt aggactaaaa gtgatgttta acttcaaaat gtttaaaatt ttgttcttcc
     8461 cgacgttcat taagaatact aatacacttt aataattatt ttaatatttt gttctaaata
     8521 atgactttta attaaaaaag ataaaatata aaaacatcat aataactcac cagaggttaa
     8581 gaacaaaaaa acaaattaga tatctgctaa tccaatatag ttaaatcaat ctttccttgg
     8641 tataatgggt atattacata tatttcaagg accgacactc ctaccaaata tctaaaattt
     8701 accatattaa cataacatgt atataaacgt caaatcataa tcagcactaa cgtctcacgg
     8761 atcgtatatg ccgtagcgac aatctaagaa ctatgcgagg acacgctagg gccaaggcaa
     8821 aaagattcct tgattacgta agggagttag aatcattttg aataaaaaac acgctttttc
     8881 agttcgagtt tatcattatc aatactgcca tttcaaagaa tacgtaaata attaatagta
     8941 gtgattttcc taactttatt tagtcaaaaa attagccttt taattctgct gtaacccgta
     9001 catgcccaaa atagggggcg ggttacacag aatatataac atcgtaggtg tctgggtgaa
     9061 cagtttattc ttggcatcca ctaaatataa tggagcccgc tttttaagct ggcatccaga
     9121 aaaaaaaaga atcccagcac caaaatattg ttttcttcac caaccatcag ttcataggtc
     9181 cattctctta gcgcaactac agagaacagg ggcacaaaca ggcaaaaaac gggcacaacc
     9241 tcaatggagt gatgcaacct gcctggagta aatgatgaca caaggcaatt gacccacgca
     9301 tgtatctatc tcattttctt acaccttcta ttaccttctg ctctctctga tttggaaaaa
     9361 gctgaaaaaa aaggttgaaa ccagttccct gaaattattc ccctacttga ctaataagta
     9421 tataaagacg gtaggtattg attgtaattc tgtaaatcta tttcttaaac ttcttaaatt
     9481 ctacttttat agttagtctt ttttttagtt ttaaaacacc aagaacttag tttcgaataa
     9541 acacacataa acaaacaaac tagtgcgcaa gtggtttagt ggtaaaattc atcgttgcca
     9601 tcgatgagcc cccggttcga ttccgggctt gcgcacaccg caggtgtgcg gacacctgct
     9661 gcagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     9721 agtggcaccg agtcggtgct tttggccggc atggtcccag cctcctcgct ggcgccggct
     9781 gggcaacatg cttcggcatg gcgaatggga cacaggcccc ttttcctttg tcgatatcat
     9841 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa
     9901 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt
     9961 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaac gcgtgtacgc
    10021 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacaagctt acgtctcacg
    10081 gatcgtatat gccgtagcga caatctaaga actatgcgag gacacgctag cagctgtatt
    10141 cacgtagacg gataggtata gccagacatc agcagcatac ttcgggaacc gtaggccacc
    10201 gagcggccgc gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
    10261 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
    10321 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
    10381 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
    10441 ccgcgggacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
    10501 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
    10561 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
    10621 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
    10681 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
    10741 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
    10801 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
    10861 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    10921 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    10981 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    11041 actcgtgtac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    11101 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    11161 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
    11221 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
    11281 cgaaaagtgc cacctgtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
    11341 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
    11401 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
    11461 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
    11521 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
    11581 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
    11641 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
    11701 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
    11761 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
    11821 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
    11881 tatagtcctg tcgggtttcc ccacctctga cttgagcgtc gatttttgtg atgctcgtca
    11941 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
    12001 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
    12061 gcggactata tgtgaaggca tggctatggc acggcagaca ttccgccaga
//
