LOCUS       sequence-352590-       11828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing optimized Cas9 and NAT marker and sgRNA targeting
            KU70 homolog in D. hansenii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11828)
  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 11828)
  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: pDIV537
FEATURES             Location/Qualifiers
     source          1..11828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             846..4949
                     /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             846..4949
                     /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             4962..4982
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4986..5006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5010..5030
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5031..5096
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(5177..5195)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      5207..5395
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     terminator      7004..7201
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    7264..7297
                     /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(7367..7400)
                     /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(7870..7887)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7870..7886)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7985..8002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8156..8744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8236..8255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8866..9657,9658..9726))
                     /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     9489..9508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9727..9818)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9860..9879
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9885..9904
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    10111..10144
                     /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)"
     misc_RNA        11519..11594
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 catgcacatc gttcgctaaa gactttatct gtgggaataa aaattgtata acttattgcc
       61 aaacttactc cctagtacgt ctctgttatt atcgcgtgca ttcagcaatt ccaaaatccc
      121 attagagggg ttcttcacga ttatctattt gatcactctc gcctaaacta taaatatagc
      181 taccttttta tcaatataca tgtacttccg aacatcggcc cttatctgtt attaatctat
      241 taataaaccc ttaacaattt cctgttgctc cttgtaaacc agtcctatat gtccaacttg
      301 cccggattgg tctgatcttc cgatctaaca atcacgtctt actaatatta taatgccttg
      361 gtaacacgtc tcgcgtttac gcgtatgccc tttaggaatt ttgtaaacaa actgccccgt
      421 agattacaaa caagagagtg aattccaccc tcaactatag acctaggagc catttttcta
      481 tataactact atctgtataa ggatggggcc tcctgggaca tgttcaattg aattttacgc
      541 ttcgaaccat acacataaca cgtaaaaagc tctaatattg ggcttcttaa atggaaagtt
      601 gtagtataga ttttacgcga tgatccaatt cgcgttttgt gtagggaatt gcttctacat
      661 caagcaaaat ttacaatgct tttatgtata cggccgtaga cggaatagac aacattgtat
      721 aatcaggaat tgatataaaa tagttgatat cccactattc taataacttc attttttatt
      781 aattagtatt tgataatata aaatttcatt agttgctttc aaacattatt cagacaatct
      841 gtcatatgga taaaaagtat agtattggtt tagatattgg tactaactct gtgggttggg
      901 cagttatcac tgacgaatat aaagttccat caaagaaatt taaggtgtta ggtaacactg
      961 acagacactc aataaaaaag aatcttatcg gtgctctttt gttcgactcc ggtgaaactg
     1021 ccgaggctac acgtttaaaa agaacagcaa gaagaagata tacccgtaga aaaaatagaa
     1081 tatgttattt acaagaaatc ttttctaatg aaatggctaa agttgatgat tcctttttcc
     1141 atagattgga agagtcattt ttggttgaag aagacaaaaa gcatgagaga catccaatct
     1201 ttgggaatat agttgatgaa gtggcttacc atgaaaaata tcctaccatt tatcatttaa
     1261 gaaagaaatt ggtagattca actgataaag ctgaccttag attaatctat ttagcacttg
     1321 cccatatgat taaatttaga ggtcattttt tgattgaagg tgatttgaac ccagataatt
     1381 ctgacgtgga taaattattt attcaattag tccaaaccta caaccaatta tttgaggaaa
     1441 atccaattaa tgctagtggt gtcgatgcca aagctatatt atcagccaga ttatcaaaat
     1501 ctagacgttt ggaaaatttg attgcccaat tgccaggaga aaaaaagaat ggattatttg
     1561 gaaacttgat cgcattatca ttgggtttga caccaaattt taaatctaat tttgatttag
     1621 ctgaagatgc taaattacaa ttatcaaaag acacctatga cgacgatttg gacaatttac
     1681 ttgctcaaat tggtgatcaa tatgcagatt tgttcttagc tgctaaaaac ttatctgatg
     1741 ctattttgtt gtctgatatt ttgagagtga acacagaaat aaccaaagct ccattatcag
     1801 catctatgat caaacgttat gatgaacacc atcaggattt gactttattg aaagctttgg
     1861 tgagacaaca attgccagag aagtataaag aaatcttttt cgatcaatct aaaaacgggt
     1921 atgcaggtta tattgatggg ggtgcctccc aagaggaatt ttacaaattt ataaaaccta
     1981 ttttagaaaa gatggatggg actgaggaac ttttggtcaa attgaacaga gaagatttgt
     2041 tacgtaaaca gagaactttt gataatggta gtatacctca ccaaattcat ttgggtgagt
     2101 tgcatgcaat tttaagaaga caagaagatt tttatccatt tttaaaagat aatagagaaa
     2161 aaatcgagaa aattttaacc tttagaattc catactatgt tgggcctttg gctagaggta
     2221 attcaagatt tgcctggatg acacgtaaat cagaagaaac tattacccct tggaattttg
     2281 aagaggttgt tgataaagga gcatcagcac agagttttat tgaaagaatg accaatttcg
     2341 ataaaaactt accaaatgaa aaagttttac caaaacattc cttgttatac gaatatttta
     2401 ctgtttacaa tgaacttaca aaggttaaat atgttactga aggtatgcgt aagccagcct
     2461 ttttatctgg agaacagaaa aaggcaatag ttgatttatt gtttaaaaca aatagaaaag
     2521 ttactgttaa acaattaaaa gaagattact ttaagaaaat tgaatgtttt gattcagttg
     2581 aaatcagtgg tgttgaagac agatttaatg ctagtttagg aacttaccat gatttactta
     2641 aaattatcaa agataaagat ttcttggata acgaagaaaa tgaagacatt ttagaagaca
     2701 ttgttttaac cttaacttta ttcgaagata gagagatgat tgaagaacgt ttgaagactt
     2761 atgcacattt gtttgacgat aaagtgatga aacagttgaa aagaagacgt tatactggat
     2821 ggggtagatt gtctcgtaaa ttgatcaatg gaattagaga taaacaaagt ggtaaaacta
     2881 tcttggactt tttgaaatct gacggatttg ctaatagaaa tttcatgcaa ttgatccacg
     2941 acgatagttt gacatttaaa gaagacatcc aaaaggccca agtgagtggg caaggtgatt
     3001 cattacatga acatattgca aatttagccg gatctcctgc tattaagaaa gggatattac
     3061 aaactgttaa agttgtggat gaattagtga aagtaatggg aagacataaa cctgaaaaca
     3121 ttgtcattga gatggcaaga gaaaatcaaa ctacacaaaa aggacagaaa aatagtagag
     3181 aacgtatgaa aagaatagaa gagggtatta aagaattggg tagtcaaata ttgaaagaac
     3241 acccagtgga aaatacccag ttgcaaaatg aaaaattata tctttactac cttcaaaatg
     3301 gacgtgatat gtatgttgat caggaattag atataaatag actttcagat tatgatgtag
     3361 atcatatagt tccacaatct ttcttgaaag atgattccat agacaataaa gtattaacta
     3421 gaagtgataa aaatagaggt aaaagtgata atgtcccaag tgaggaagtc gtcaaaaaga
     3481 tgaaaaatta ctggcgtcaa cttttgaatg ctaaattaat tactcaaaga aaatttgata
     3541 atttgactaa agcagaaaga ggtgggcttt ctgaattaga taaagccggg ttcattaaaa
     3601 gacaattggt cgaaactaga caaattacta aacatgttgc ccaaatttta gattcccgta
     3661 tgaacactaa gtatgacgaa aatgataagt taatacgtga ggttaaagtc attactttaa
     3721 aatcaaaact tgtctctgat ttcagaaagg atttccaatt ctataaagtt agagaaatta
     3781 ataattatca tcatgctcat gatgcatatt tgaatgctgt agttggaact gctttaatca
     3841 agaaataccc taaattagaa tctgaatttg tatatggtga ttacaaagtc tatgatgtta
     3901 gaaagatgat tgctaaatca gaacaagaaa ttggtaaagc tacagctaaa tacttctttt
     3961 actctaacat tatgaatttc tttaaaacag aaattacttt ggcaaacggt gaaattagaa
     4021 aaagacctct tattgaaaca aatggtgaga ctggagagat agtttgggac aaagggcgtg
     4081 atttcgctac tgttagaaaa gttttatcaa tgccacaagt taacattgta aagaaaacag
     4141 aggttcaaac tggtggtttc tcaaaagaaa gtattttgcc taaaagaaat agtgataaat
     4201 tgattgccag aaaaaaggat tgggatccaa agaaatatgg tggtttcgac tcaccaaccg
     4261 tagcctattc tgttttggtt gtggcaaagg ttgaaaaggg taaaagtaaa aagcttaaat
     4321 cagtaaaaga acttttgggt attacaataa tggaaagaag ttcctttgaa aagaacccta
     4381 ttgatttttt ggaagctaaa ggttataagg aagtaaagaa ggacttaata atcaaattgc
     4441 ctaaatattc tttatttgaa ttagaaaatg ggagaaaaag aatgttggct tctgctggag
     4501 aattgcaaaa gggtaatgaa ttagcattgc cttccaaata tgttaacttc ttgtatttag
     4561 cttcacacta tgaaaagttg aaagggtcac cagaagataa cgagcaaaaa caattatttg
     4621 ttgaacaaca caaacactac ttagatgaga ttatagaaca aattagtgaa ttcagtaaaa
     4681 gagtgatatt agctgatgca aatttagata aagttttgtc agcctataac aaacatagag
     4741 ataagccaat tagagaacaa gcagaaaaca ttattcactt atttaccctt accaatttag
     4801 gagcacctgc tgctttcaag tattttgata caacaattga tcgtaaaaga tatacctcaa
     4861 caaaagaagt cttagacgcc accttaattc atcaatcaat cactggattg tatgagacaa
     4921 gaattgattt gtctcaattg ggtggtgatg aaggggctga tcctaagaag aaaagaaaag
     4981 ttgatccaaa gaaaaagcgt aaggtggatc ctaagaaaaa gagaaaggtt gactacaaag
     5041 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtgat
     5101 aatgactgca gagatccatc gacctgccgc caagctaatt ccgggcgaat ttctgtcgag
     5161 tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg
     5221 aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt
     5281 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt
     5341 acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaatcgc
     5401 gtgcattcaa cgctgcaggt cgacaaccct taatataact tggaaagcaa gcaatcgtag
     5461 ccccagcaaa gcactatagt aggtgtttgc tctttttatt tcctttgctc tacgcactgt
     5521 aatacaggat tgatgaaagc agtatagaaa atcgcctcga agcctcgtga atcagtagat
     5581 ctatttacat tgttcgtctc gtgcggtttt tcaccagtgt catgagcgag cgaacttggt
     5641 cgacatcatt aaaaaaaact gcggctttcg tgtcagagtc gctgtgtatc acttcgacgt
     5701 aggtaatcag ctcttaagct tgaagtgtgg atcggaattt tcttctcctc cttcattagt
     5761 catcttccat gaaagagtga cttttgactc acacgaagga aaatgtgcgc aaattacaaa
     5821 tcacctacaa gccacaagac acattcatca aaggcaccaa acaagacatt caagtggacc
     5881 atgataaaag ccaaggtttg aaggcatatt ctgtatactt caatcttggt ttgctcggcc
     5941 caagtcccgg cgattagcgg cgcccggtgg cccggctgga tagatttcaa ttggcacacg
     6001 cacaattatc tccacaattg cattgcatac acctttacac cacaactcgg cgacctcggc
     6061 cagcctctga aaacacatag ccaaaaacga agaagggaca ggcattttcc cttcattgcc
     6121 tagtatggca taatacgtca ccaaaccatg cattatggat tgttaagagc taatcttcct
     6181 ggtgaaaaac cgccacaaat tgaccatgac agtgcttgct cggagttttc agcggagggc
     6241 gtgcgcccgg gcaaacatag tgcattgtga tgtaggcggc aaatatatgc cggatacgcg
     6301 cttatggagc ggattgcccc ggcggagaaa agtatataag tagccagcgc aaggccccca
     6361 tttctagaga atttttccct tcaattgatt acaacttcaa ttaactacaa acaaaacaaa
     6421 caaccatggg aaccaccttg gatgacactg cttatcgata ccgaacatct gtacctggtg
     6481 atgctgaagc tatcgaagcc cttgacgggt cattcacaac tgacaccgtc tttagagtta
     6541 cagctactgg ggatgggttt acattgcgag aagtaccagt agatcctcct ttgaccaaag
     6601 tattcccaga tgacgaaagt gacgatgaat ccgatgacgg tgaggacggt gacccagact
     6661 caagaacatt tgtcgcttat ggagacgacg gtgatttggc cggtttcgta gttgtatcct
     6721 attctggatg gaaccgaaga cttactgtcg aggatataga ggttgcccca gaacatcgtg
     6781 ggcacggtgt tgggcgagcc cttatgggat tagctacaga atttgcccgt gaacgtgggg
     6841 ctggacattt atggttagaa gtcaccaatg ttaatgcacc agctatacat gcatatcgaa
     6901 gaatggggtt cacactttgt ggattggata cagctttata cgatggtaca gcctcagacg
     6961 gagaacaagc tttatacatg tctatgcctt gtccataatc agtactgaca ataaaaagat
     7021 tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta
     7081 atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa
     7141 gttaagtgcg cagaaagtaa tatcatgcgt taatcgtatg tgaatgctgg tcgctatact
     7201 gctgtcgatt cgatactaac gccgccatcc agtgtcgaaa acgagctctc gagaaccctt
     7261 aatataactt cgtataatgt atgctatacg aagttattag gtgatatcag atccactagt
     7321 ggccattaag acctcagcgc ggccgcaaat ttaaataaaa tgaagtgaag ttcctatact
     7381 ttctagagaa taggaacttc tatagtgagt cgaataaggg cgacacaaaa tttattctaa
     7441 atgcataata aatactgata acatcttata gtttgtatta tattttgtat tatcgttgac
     7501 atgtataatt ttgatatcaa aaactgattt tccctttatt attttcgaga tttattttct
     7561 taattctctt taacaaacta gaaatattgt atatacaaaa aatcataaat aatagatgaa
     7621 tagtttaatt ataggtgttc atcaatcgaa aaagcaacgt atcttattta aagtgcgttg
     7681 cttttttctc atttataagg ttaaataatt ctcatatatc aagcaaagtg acaggcgccc
     7741 ttaaatattc tgacaaatgc tctttcccta aactcccccc ataaaaaaac ccgccgaagc
     7801 gggtttttac gttatttgcg gattaacgat tactcgttat cagaaccgcc cagggggccc
     7861 gagcttaaga ctggccgtcg ttttacaaca cagaaagagt ttgtagaaac gcaaaaaggc
     7921 catccgtcag gggccttctg cttagtttga tgcctggcag ttccctactc tcgccttccg
     7981 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     8041 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     8101 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     8161 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     8221 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     8281 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     8341 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     8401 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     8461 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     8521 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tgggctaact
     8581 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     8641 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     8701 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8761 tttctacggg gtctgacgct cagtggaacg acgcgcgcgt aactcacgtt aagggatttt
     8821 ggtcatgagc ttgcgccgtc ccgtcaagtc agcgtaatgc tctgcttacc aatgcttaat
     8881 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8941 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagcg ctgcgatgat
     9001 accgcgagaa ccacgctcac cggctccgga tttatcagca ataaaccagc cagccggaag
     9061 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     9121 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccatcgc
     9181 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     9241 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     9301 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     9361 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     9421 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     9481 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9541 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     9601 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     9661 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     9721 actcatattc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9781 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggtcagtg ttacaaccaa
     9841 ttaaccaatt ctgaacatta tcgcgagccc atttatacct gaatatggct cataacaccc
     9901 cttgtttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     9961 tgaaacgccg tagcgccgat ggtagtgtgg ggactcccca tgcgagagta gggaactgcc
    10021 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgccc gggctaatta
    10081 tggggtgtcg cccttattcg actctatagt gaagttccta ttctctagaa agtataggaa
    10141 cttctgaagt ggggatttaa atgcggccgc gctgagggtt taatgggctt agttagcctt
    10201 gcttgtgact ttgcagcttt atccatgaac ggatcatacg attgtatttc tgaatttgtc
    10261 atagtgactt taagttgttc tttatttggt atgattctat tttttaggac tttctaaaaa
    10321 atcaatttaa attttggaaa accatttgtg gtttttctta aatgattgca aaattattaa
    10381 aaaatcgtta aaaaaatatt cagtgtaaaa agactagcat tcatgatctc aacatctttg
    10441 gataatatca gaatgagaaa gaacagatac gcagtacgtt ttttggtgag ctctttgcac
    10501 ttctttagtt ctttccatca atatcagttt tttaaacttt taggactaaa agtgatgttt
    10561 aacttcaaaa tgtttaaaat tttgttcttc ccgacgttca ttaagaatac taatacactt
    10621 taataattat tttaatattt tgttctaaat aatgactttt aattaaaaaa gataaaatat
    10681 aaaaacatca taataactca ccagaggtta agaacaaaaa aacaaattag atatctgcta
    10741 atccaatata gttaaatcaa tctttccttg gtataatggg tatattacat atatttcaag
    10801 gaccgacact cctaccaaat atctaaaatt taccatatta acataacatg tatataaacg
    10861 tcaaatcata atcagcacta gcggaatgcg tgcgatatcc caagccacca taataaatat
    10921 acattcatag tgccatctcg gcccactgca tttatccact gtatatgcaa tatacgcacg
    10981 gtatctcgag aaacagtcgt aggaggcaac ctgacgtacc cattcttcgt ttacttcgct
    11041 tccaacatac tcagacgcca tcatagcgac tgcaaaatgt tctcttagcc ttcatacccc
    11101 tattcgtact aatgttcata aaaaaaaaca ccaaaatagc caatctggaa aaaacatgtg
    11161 tccctttttg atttctattc tacaactaca tgttgcagcc atattgacat agtctctaac
    11221 cagaatacct gccattagga cctcctaaag agggaatagg gtatagccct atccgggtaa
    11281 acgcgatttc agttcggctg aaattttttt ttggccacga caccattgtc gaatgttctg
    11341 tggaaaaatt tataaggaag taaattgaaa aagttcttct tcttctcttt tttcagtaca
    11401 gtatactatt attatacatt aaacaaagcg caaatggttt agtggtaaaa tccaacgttg
    11461 ccatcgttgg gcccccggtt cgattccggg tttgcgcaac tgcttcttca ttcaattcgt
    11521 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
    11581 caccgagtcg gtgctttgcg caaatggttt agtggtaaaa tccaacgttg ccatcgttgg
    11641 gcccccggtt cgattccggg tttgcgcagc tgattgatat agatttatat tagttgcttg
    11701 tcttatattt agttttttga atacatatga attttaaatt tagttgtaga cctttggaca
    11761 ggctttaatg ctgtctgtat cggtatatat cttggaccat gtggtattaa tctaatcgaa
    11821 tgtgtgag
//
