LOCUS       sequence-352589-       12820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing optimized Cas9 and NAT marker and sgRNA targeting
            ADE2 locus in D. hansenii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12820)
  AUTHORS   Strucko T, Andersen NL, Mahler MR, Martinez JL, Mortensen UH
  TITLE     A CRISPR/Cas9 method facilitates efficient oligo-mediated gene 
            editing in Debaryomyces hansenii.
  JOURNAL   Synth Biol (Oxf). 2021 Oct 12;6(1):ysab031. doi: 
            10.1093/synbio/ysab031. eCollection 2021.
  PUBMED    34746438
REFERENCE   2  (bases 1 to 12820)
  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: "Synth Biol 
            (Oxf)."; date: "2021-10-12"; volume: "6(1):ysab031. doi"; pages: " 
            10.1093/synbio/ysab031. eCollection 2021"
COMMENT     Sequence Label: pDIV494
FEATURES             Location/Qualifiers
     source          1..12820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1643..1718
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             3410..7513
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3410..7513
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7526..7546
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7550..7570
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7574..7594
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7595..7660
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(7741..7759)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      7771..7959
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     terminator      9568..9765
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    9828..9861
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(9931..9964)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(10434..10451)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10434..10450)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10549..10566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10720..11308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10800..10819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11430..12221,12222..12290))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     12053..12072
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12291..12382)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     12424..12443
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     12449..12468
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    12675..12708
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 tgtcatagtg actttaagtt gttctttatt tggtatgatt ctatttttta ggactttcta
       61 aaaaatcaat ttaaattttg gaaaaccatt tgtggttttt cttaaatgat tgcaaaatta
      121 ttaaaaaatc gttaaaaaaa tattcagtgt aaaaagacta gcattcatga tctcaacatc
      181 tttggataat atcagaatga gaaagaacag atacgcagta cgttttttgg tgagctcttt
      241 gcacttcttt agttctttcc atcaatatca gttttttaaa cttttaggac taaaagtgat
      301 gtttaacttc aaaatgttta aaattttgtt cttcccgacg ttcattaaga atactaatac
      361 actttaataa ttattttaat attttgttct aaataatgac ttttaattaa aaaagataaa
      421 atataaaaac atcataataa ctcaccagag gttaagaaca aaaaaacaaa ttagatatct
      481 gctaatccaa tatagttaaa tcaatctttc cttggtataa tgggtatatt acatatattt
      541 caaggaccga cactcctacc aaatatctaa aatttaccat attaacataa catgtatata
      601 aacgtcaaat cataatcagc actagcggaa tgcgtgcgat gcatcaagct ggtctcattg
      661 acttggcgtc cagtaacggc attcgtttac cagcctccat tataaacgct tctagggctg
      721 ttccagaaga tgacgaagag gaaaacagcc tggccatggt tcctgctcta gactgggatc
      781 tttggggccc tttgattttc tccttggctt attcggtcac tttggggttc gctgcaccca
      841 actctcagac caatatggtc ttctcgggaa cgttttcttt catctgggtg ttttacttaa
      901 ttggtggctt gaacatccaa cttttagggg gtacaatctc cttcttgtct gccatcagtg
      961 catcaggata ttccatgttt cctgtcgtgg ttggtgccgt ggtgaacact ttagcaatca
     1021 aatggagatg gttgcgtttg ctcattatgt gttttctcac aacttggagc atttatgccg
     1081 caggaatgag cctacgttgt ctgggagtat tgcctggaag agtatttttg gccatgtatc
     1141 cggttgcttt gatgtacacc gttttggcat atcttaccgt cataacataa ttgtaacccg
     1201 aggtttctag cacttaatac atgatgtact aaaaggtgcg agtaattacg aggctaccca
     1261 taactttaat gaattaagct tgcgacatat ggcattattt attgctgtgc aagccatgaa
     1321 tatgtaaatg taaccaaata tacctagtga gttgtgcaat catgtcttct tcagtacacg
     1381 taaactacca acatactttc agtgaagaaa acacaataac ccaaacaggc acaatcacag
     1441 acgcagctaa aaccacctca tatcttcgca gacatttctt catgaaatca cttacatttt
     1501 tcactacagg ttgctcatac tcgctgagaa aaaaaaatgt tgttatttca tttgctgtgg
     1561 cccagtcggg acgcgcaatg cgcgatccgg catgcggagg ttcgattcct agataaatga
     1621 tcagctaagc agattaatgc atgttttaga gctagaaata gcaagttaaa ataaggctag
     1681 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttctcga gtttttttat
     1741 cgagtgttta aggataatga taactgaaga gaagaattag ttttgccgcc accgcgggtt
     1801 tgcctctgat taaataaaaa aaagctggtg cttttttttc ttttatagga acatcttgaa
     1861 tatatgaact aattaaatga taatttttta cccatcttta ctcttaatca ctgagctgca
     1921 gtcaaagaaa aagggataca gcacctggtg aagagatgaa cggagactaa cttagacgcg
     1981 ttgattcttt tcaattgcac attttattaa tcgatgctaa cgtctattta catatattct
     2041 ttagagatat tatctagggc ttcaaataat ctctggacag caataaaagt ctcttcaaaa
     2101 gtattgtata acggcaatgg ggctaatctg attacatctg gtcttctttc gtcacagatt
     2161 atagcatgat catgcaagta cgcattaact cgttccatga cgttcttgcc cttttcatcg
     2221 aaatgcggtt gaaacataat ggacaattga catcctcttt cagctggatt caaaggagtt
     2281 aaaattttaa acccaaattt ggagtttgat gtactggatt gtggtatgta atacttggaa
     2341 ttcgtcaata gatcctgtaa aaattgagtc aaagcaacac ttttttcacg aagtttagat
     2401 actccaccca ctttagcata cacttccaat gacgacttca cagcaacaac atcaagaaca
     2461 gaaggatttg actgtctgta agaaagagcc gagtttattg gatcaaactc ttctaacatt
     2521 ttgaatcgtt cttgggagtt attgccccac caaccagcta gtctaggaac gaaactgctc
     2581 ttcttgttct ctatagtgta tttttcatgc acaaaaattc cacctatggc tccaggtccc
     2641 gagtttaaat atttatcgcg tgcattcagc aattccaaaa tcccattaga ggggttcttc
     2701 acgattatct atttgatcac tctcgcctaa actataaata tagctacctt tttatcaata
     2761 tacatgtact tccgaacatc ggcccttatc tgttattaat ctattaataa acccttaaca
     2821 atttcctgtt gctccttgta aaccagtcct atatgtccaa cttgcccgga ttggtctgat
     2881 cttccgatct aacaatcacg tcttactaat attataatgc cttggtaaca cgtctcgcgt
     2941 ttacgcgtat gccctttagg aattttgtaa acaaactgcc ccgtagatta caaacaagag
     3001 agtgaattcc accctcaact atagacctag gagccatttt tctatataac tactatctgt
     3061 ataaggatgg ggcctcctgg gacatgttca attgaatttt acgcttcgaa ccatacacat
     3121 aacacgtaaa aagctctaat attgggcttc ttaaatggaa agttgtagta tagattttac
     3181 gcgatgatcc aattcgcgtt ttgtgtaggg aattgcttct acatcaagca aaatttacaa
     3241 tgcttttatg tatacggccg tagacggaat agacaacatt gtataatcag gaattgatat
     3301 aaaatagttg atatcccact attctaataa cttcattttt tattaattag tatttgataa
     3361 tataaaattt cattagttgc tttcaaacat tattcagaca atctgtcata tggataaaaa
     3421 gtatagtatt ggtttagata ttggtactaa ctctgtgggt tgggcagtta tcactgacga
     3481 atataaagtt ccatcaaaga aatttaaggt gttaggtaac actgacagac actcaataaa
     3541 aaagaatctt atcggtgctc ttttgttcga ctccggtgaa actgccgagg ctacacgttt
     3601 aaaaagaaca gcaagaagaa gatatacccg tagaaaaaat agaatatgtt atttacaaga
     3661 aatcttttct aatgaaatgg ctaaagttga tgattccttt ttccatagat tggaagagtc
     3721 atttttggtt gaagaagaca aaaagcatga gagacatcca atctttggga atatagttga
     3781 tgaagtggct taccatgaaa aatatcctac catttatcat ttaagaaaga aattggtaga
     3841 ttcaactgat aaagctgacc ttagattaat ctatttagca cttgcccata tgattaaatt
     3901 tagaggtcat tttttgattg aaggtgattt gaacccagat aattctgacg tggataaatt
     3961 atttattcaa ttagtccaaa cctacaacca attatttgag gaaaatccaa ttaatgctag
     4021 tggtgtcgat gccaaagcta tattatcagc cagattatca aaatctagac gtttggaaaa
     4081 tttgattgcc caattgccag gagaaaaaaa gaatggatta tttggaaact tgatcgcatt
     4141 atcattgggt ttgacaccaa attttaaatc taattttgat ttagctgaag atgctaaatt
     4201 acaattatca aaagacacct atgacgacga tttggacaat ttacttgctc aaattggtga
     4261 tcaatatgca gatttgttct tagctgctaa aaacttatct gatgctattt tgttgtctga
     4321 tattttgaga gtgaacacag aaataaccaa agctccatta tcagcatcta tgatcaaacg
     4381 ttatgatgaa caccatcagg atttgacttt attgaaagct ttggtgagac aacaattgcc
     4441 agagaagtat aaagaaatct ttttcgatca atctaaaaac gggtatgcag gttatattga
     4501 tgggggtgcc tcccaagagg aattttacaa atttataaaa cctattttag aaaagatgga
     4561 tgggactgag gaacttttgg tcaaattgaa cagagaagat ttgttacgta aacagagaac
     4621 ttttgataat ggtagtatac ctcaccaaat tcatttgggt gagttgcatg caattttaag
     4681 aagacaagaa gatttttatc catttttaaa agataataga gaaaaaatcg agaaaatttt
     4741 aacctttaga attccatact atgttgggcc tttggctaga ggtaattcaa gatttgcctg
     4801 gatgacacgt aaatcagaag aaactattac cccttggaat tttgaagagg ttgttgataa
     4861 aggagcatca gcacagagtt ttattgaaag aatgaccaat ttcgataaaa acttaccaaa
     4921 tgaaaaagtt ttaccaaaac attccttgtt atacgaatat tttactgttt acaatgaact
     4981 tacaaaggtt aaatatgtta ctgaaggtat gcgtaagcca gcctttttat ctggagaaca
     5041 gaaaaaggca atagttgatt tattgtttaa aacaaataga aaagttactg ttaaacaatt
     5101 aaaagaagat tactttaaga aaattgaatg ttttgattca gttgaaatca gtggtgttga
     5161 agacagattt aatgctagtt taggaactta ccatgattta cttaaaatta tcaaagataa
     5221 agatttcttg gataacgaag aaaatgaaga cattttagaa gacattgttt taaccttaac
     5281 tttattcgaa gatagagaga tgattgaaga acgtttgaag acttatgcac atttgtttga
     5341 cgataaagtg atgaaacagt tgaaaagaag acgttatact ggatggggta gattgtctcg
     5401 taaattgatc aatggaatta gagataaaca aagtggtaaa actatcttgg actttttgaa
     5461 atctgacgga tttgctaata gaaatttcat gcaattgatc cacgacgata gtttgacatt
     5521 taaagaagac atccaaaagg cccaagtgag tgggcaaggt gattcattac atgaacatat
     5581 tgcaaattta gccggatctc ctgctattaa gaaagggata ttacaaactg ttaaagttgt
     5641 ggatgaatta gtgaaagtaa tgggaagaca taaacctgaa aacattgtca ttgagatggc
     5701 aagagaaaat caaactacac aaaaaggaca gaaaaatagt agagaacgta tgaaaagaat
     5761 agaagagggt attaaagaat tgggtagtca aatattgaaa gaacacccag tggaaaatac
     5821 ccagttgcaa aatgaaaaat tatatcttta ctaccttcaa aatggacgtg atatgtatgt
     5881 tgatcaggaa ttagatataa atagactttc agattatgat gtagatcata tagttccaca
     5941 atctttcttg aaagatgatt ccatagacaa taaagtatta actagaagtg ataaaaatag
     6001 aggtaaaagt gataatgtcc caagtgagga agtcgtcaaa aagatgaaaa attactggcg
     6061 tcaacttttg aatgctaaat taattactca aagaaaattt gataatttga ctaaagcaga
     6121 aagaggtggg ctttctgaat tagataaagc cgggttcatt aaaagacaat tggtcgaaac
     6181 tagacaaatt actaaacatg ttgcccaaat tttagattcc cgtatgaaca ctaagtatga
     6241 cgaaaatgat aagttaatac gtgaggttaa agtcattact ttaaaatcaa aacttgtctc
     6301 tgatttcaga aaggatttcc aattctataa agttagagaa attaataatt atcatcatgc
     6361 tcatgatgca tatttgaatg ctgtagttgg aactgcttta atcaagaaat accctaaatt
     6421 agaatctgaa tttgtatatg gtgattacaa agtctatgat gttagaaaga tgattgctaa
     6481 atcagaacaa gaaattggta aagctacagc taaatacttc ttttactcta acattatgaa
     6541 tttctttaaa acagaaatta ctttggcaaa cggtgaaatt agaaaaagac ctcttattga
     6601 aacaaatggt gagactggag agatagtttg ggacaaaggg cgtgatttcg ctactgttag
     6661 aaaagtttta tcaatgccac aagttaacat tgtaaagaaa acagaggttc aaactggtgg
     6721 tttctcaaaa gaaagtattt tgcctaaaag aaatagtgat aaattgattg ccagaaaaaa
     6781 ggattgggat ccaaagaaat atggtggttt cgactcacca accgtagcct attctgtttt
     6841 ggttgtggca aaggttgaaa agggtaaaag taaaaagctt aaatcagtaa aagaactttt
     6901 gggtattaca ataatggaaa gaagttcctt tgaaaagaac cctattgatt ttttggaagc
     6961 taaaggttat aaggaagtaa agaaggactt aataatcaaa ttgcctaaat attctttatt
     7021 tgaattagaa aatgggagaa aaagaatgtt ggcttctgct ggagaattgc aaaagggtaa
     7081 tgaattagca ttgccttcca aatatgttaa cttcttgtat ttagcttcac actatgaaaa
     7141 gttgaaaggg tcaccagaag ataacgagca aaaacaatta tttgttgaac aacacaaaca
     7201 ctacttagat gagattatag aacaaattag tgaattcagt aaaagagtga tattagctga
     7261 tgcaaattta gataaagttt tgtcagccta taacaaacat agagataagc caattagaga
     7321 acaagcagaa aacattattc acttatttac ccttaccaat ttaggagcac ctgctgcttt
     7381 caagtatttt gatacaacaa ttgatcgtaa aagatatacc tcaacaaaag aagtcttaga
     7441 cgccacctta attcatcaat caatcactgg attgtatgag acaagaattg atttgtctca
     7501 attgggtggt gatgaagggg ctgatcctaa gaagaaaaga aaagttgatc caaagaaaaa
     7561 gcgtaaggtg gatcctaaga aaaagagaaa ggttgactac aaagaccatg acggtgatta
     7621 taaagatcat gacatcgact acaaggatga cgatgacaag tgataatgac tgcagagatc
     7681 catcgacctg ccgccaagct aattccgggc gaatttctgt cgagtcatgt aattagttat
     7741 gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag
     7801 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
     7861 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
     7921 actgaaaacc ttgcttgaga aggttttggg acgctcgaaa tcgcgtgcat tcaacgctgc
     7981 aggtcgacaa cccttaatat aacttggaaa gcaagcaatc gtagccccag caaagcacta
     8041 tagtaggtgt ttgctctttt tatttccttt gctctacgca ctgtaataca ggattgatga
     8101 aagcagtata gaaaatcgcc tcgaagcctc gtgaatcagt agatctattt acattgttcg
     8161 tctcgtgcgg tttttcacca gtgtcatgag cgagcgaact tggtcgacat cattaaaaaa
     8221 aactgcggct ttcgtgtcag agtcgctgtg tatcacttcg acgtaggtaa tcagctctta
     8281 agcttgaagt gtggatcgga attttcttct cctccttcat tagtcatctt ccatgaaaga
     8341 gtgacttttg actcacacga aggaaaatgt gcgcaaatta caaatcacct acaagccaca
     8401 agacacattc atcaaaggca ccaaacaaga cattcaagtg gaccatgata aaagccaagg
     8461 tttgaaggca tattctgtat acttcaatct tggtttgctc ggcccaagtc ccggcgatta
     8521 gcggcgcccg gtggcccggc tggatagatt tcaattggca cacgcacaat tatctccaca
     8581 attgcattgc atacaccttt acaccacaac tcggcgacct cggccagcct ctgaaaacac
     8641 atagccaaaa acgaagaagg gacaggcatt ttcccttcat tgcctagtat ggcataatac
     8701 gtcaccaaac catgcattat ggattgttaa gagctaatct tcctggtgaa aaaccgccac
     8761 aaattgacca tgacagtgct tgctcggagt tttcagcgga gggcgtgcgc ccgggcaaac
     8821 atagtgcatt gtgatgtagg cggcaaatat atgccggata cgcgcttatg gagcggattg
     8881 ccccggcgga gaaaagtata taagtagcca gcgcaaggcc cccatttcta gagaattttt
     8941 cccttcaatt gattacaact tcaattaact acaaacaaaa caaacaacca tgggaaccac
     9001 cttggatgac actgcttatc gataccgaac atctgtacct ggtgatgctg aagctatcga
     9061 agcccttgac gggtcattca caactgacac cgtctttaga gttacagcta ctggggatgg
     9121 gtttacattg cgagaagtac cagtagatcc tcctttgacc aaagtattcc cagatgacga
     9181 aagtgacgat gaatccgatg acggtgagga cggtgaccca gactcaagaa catttgtcgc
     9241 ttatggagac gacggtgatt tggccggttt cgtagttgta tcctattctg gatggaaccg
     9301 aagacttact gtcgaggata tagaggttgc cccagaacat cgtgggcacg gtgttgggcg
     9361 agcccttatg ggattagcta cagaatttgc ccgtgaacgt ggggctggac atttatggtt
     9421 agaagtcacc aatgttaatg caccagctat acatgcatat cgaagaatgg ggttcacact
     9481 ttgtggattg gatacagctt tatacgatgg tacagcctca gacggagaac aagctttata
     9541 catgtctatg ccttgtccat aatcagtact gacaataaaa agattcttgt tttcaagaac
     9601 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
     9661 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
     9721 gtaatatcat gcgttaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac
     9781 taacgccgcc atccagtgtc gaaaacgagc tctcgagaac ccttaatata acttcgtata
     9841 atgtatgcta tacgaagtta ttaggtgata tcagatccac tagtggccat taagacctca
     9901 gcgcggccgc aaatttaaat aaaatgaagt gaagttccta tactttctag agaataggaa
     9961 cttctatagt gagtcgaata agggcgacac aaaatttatt ctaaatgcat aataaatact
    10021 gataacatct tatagtttgt attatatttt gtattatcgt tgacatgtat aattttgata
    10081 tcaaaaactg attttccctt tattattttc gagatttatt ttcttaattc tctttaacaa
    10141 actagaaata ttgtatatac aaaaaatcat aaataataga tgaatagttt aattataggt
    10201 gttcatcaat cgaaaaagca acgtatctta tttaaagtgc gttgcttttt tctcatttat
    10261 aaggttaaat aattctcata tatcaagcaa agtgacaggc gcccttaaat attctgacaa
    10321 atgctctttc cctaaactcc ccccataaaa aaacccgccg aagcgggttt ttacgttatt
    10381 tgcggattaa cgattactcg ttatcagaac cgcccagggg gcccgagctt aagactggcc
    10441 gtcgttttac aacacagaaa gagtttgtag aaacgcaaaa aggccatccg tcaggggcct
    10501 tctgcttagt ttgatgcctg gcagttccct actctcgcct tccgcttcct cgctcactga
    10561 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    10621 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    10681 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    10741 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    10801 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    10861 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    10921 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    10981 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    11041 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    11101 gtatgtaggc ggtgctacag agttcttgaa gtggtgggct aactacggct acactagaag
    11161 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    11221 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    11281 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    11341 cgctcagtgg aacgacgcgc gcgtaactca cgttaaggga ttttggtcat gagcttgcgc
    11401 cgtcccgtca agtcagcgta atgctctgct taccaatgct taatcagtga ggcacctatc
    11461 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    11521 acgatacggg agggcttacc atctggcccc agcgctgcga tgataccgcg agaaccacgc
    11581 tcaccggctc cggatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    11641 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    11701 agtagttcgc cagttaatag tttgcgcaac gttgttgcca tcgctacagg catcgtggtg
    11761 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    11821 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    11881 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    11941 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    12001 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    12061 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    12121 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    12181 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    12241 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat attcttcctt
    12301 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    12361 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
    12421 attatcgcga gcccatttat acctgaatat ggctcataac accccttgtt tgcctggcgg
    12481 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
    12541 cgatggtagt gtggggactc cccatgcgag agtagggaac tgccaggcat caaataaaac
    12601 gaaaggctca gtcgaaagac tgggcctttc gcccgggcta attatggggt gtcgccctta
    12661 ttcgactcta tagtgaagtt cctattctct agaaagtata ggaacttctg aagtggggat
    12721 ttaaatgcgg ccgcgctgag ggtttaatgg gcttagttag ccttgcttgt gactttgcag
    12781 ctttatccat gaacggatca tacgattgta tttctgaatt
//
