LOCUS       sequence-353317-       11828 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 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: pDIV515
FEATURES             Location/Qualifiers
     source          1..11828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    110..143
                     /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        1518..1593
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             2673..6776
                     /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             2673..6776
                     /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             6789..6809
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6813..6833
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6837..6857
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6858..6923
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(7004..7022)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      7034..7222
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     terminator      8831..9028
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    9091..9124
                     /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(9194..9227)
                     /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(9697..9714)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9697..9713)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9812..9829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9983..10571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10063..10082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10693..11484,11485..11553))
                     /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     11316..11335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11554..11645)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     11687..11706
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     11712..11731
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
       61 ctttcgcccg ggctaattat ggggtgtcgc ccttattcga ctctatagtg aagttcctat
      121 tctctagaaa gtataggaac ttctgaagtg gggatttaaa tgcggccgcg ctgagggttt
      181 aatgggctta gttagccttg cttgtgactt tgcagcttta tccatgaacg gatcatacga
      241 ttgtatttct gaatttgtca tagtgacttt aagttgttct ttatttggta tgattctatt
      301 ttttaggact ttctaaaaaa tcaatttaaa ttttggaaaa ccatttgtgg tttttcttaa
      361 atgattgcaa aattattaaa aaatcgttaa aaaaatattc agtgtaaaaa gactagcatt
      421 catgatctca acatctttgg ataatatcag aatgagaaag aacagatacg cagtacgttt
      481 tttggtgagc tctttgcact tctttagttc tttccatcaa tatcagtttt ttaaactttt
      541 aggactaaaa gtgatgttta acttcaaaat gtttaaaatt ttgttcttcc cgacgttcat
      601 taagaatact aatacacttt aataattatt ttaatatttt gttctaaata atgactttta
      661 attaaaaaag ataaaatata aaaacatcat aataactcac cagaggttaa gaacaaaaaa
      721 acaaattaga tatctgctaa tccaatatag ttaaatcaat ctttccttgg tataatgggt
      781 atattacata tatttcaagg accgacactc ctaccaaata tctaaaattt accatattaa
      841 cataacatgt atataaacgt caaatcataa tcagcactag cggaatgcgt gcgatatccc
      901 aagccaccat aataaatata cattcatagt gccatctcgg cccactgcat ttatccactg
      961 tatatgcaat atacgcacgg tatctcgaga aacagtcgta ggaggcaacc tgacgtaccc
     1021 attcttcgtt tacttcgctt ccaacatact cagacgccat catagcgact gcaaaatgtt
     1081 ctcttagcct tcatacccct attcgtacta atgttcataa aaaaaaacac caaaatagcc
     1141 aatctggaaa aaacatgtgt ccctttttga tttctattct acaactacat gttgcagcca
     1201 tattgacata gtctctaacc agaatacctg ccattaggac ctcctaaaga gggaataggg
     1261 tatagcccta tccgggtaaa cgcgatttca gttcggctga aatttttttt tggccacgac
     1321 accattgtcg aatgttctgt ggaaaaattt ataaggaagt aaattgaaaa agttcttctt
     1381 cttctctttt ttcagtacag tatactatta ttatacatta aacaaagcgc aaatggttta
     1441 gtggtaaaat ccaacgttgc catcgttggg cccccggttc gattccgggt ttgcgcaagc
     1501 taagcagatt aatgcatgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     1561 tatcaacttg aaaaagtggc accgagtcgg tgctttgcgc aaatggttta gtggtaaaat
     1621 ccaacgttgc catcgttggg cccccggttc gattccgggt ttgcgcagct gattgatata
     1681 gatttatatt agttgcttgt cttatattta gttttttgaa tacatatgaa ttttaaattt
     1741 agttgtagac ctttggacag gctttaatgc tgtctgtatc ggtatatatc ttggaccatg
     1801 tggtattaat ctaatcgaat gtgtgagcat gcacatcgtt cgctaaagac tttatctgtg
     1861 ggaataaaaa ttgtataact tattgccaaa cttactccct agtacgtctc tgttattatc
     1921 gcgtgcattc agcaattcca aaatcccatt agaggggttc ttcacgatta tctatttgat
     1981 cactctcgcc taaactataa atatagctac ctttttatca atatacatgt acttccgaac
     2041 atcggccctt atctgttatt aatctattaa taaaccctta acaatttcct gttgctcctt
     2101 gtaaaccagt cctatatgtc caacttgccc ggattggtct gatcttccga tctaacaatc
     2161 acgtcttact aatattataa tgccttggta acacgtctcg cgtttacgcg tatgcccttt
     2221 aggaattttg taaacaaact gccccgtaga ttacaaacaa gagagtgaat tccaccctca
     2281 actatagacc taggagccat ttttctatat aactactatc tgtataagga tggggcctcc
     2341 tgggacatgt tcaattgaat tttacgcttc gaaccataca cataacacgt aaaaagctct
     2401 aatattgggc ttcttaaatg gaaagttgta gtatagattt tacgcgatga tccaattcgc
     2461 gttttgtgta gggaattgct tctacatcaa gcaaaattta caatgctttt atgtatacgg
     2521 ccgtagacgg aatagacaac attgtataat caggaattga tataaaatag ttgatatccc
     2581 actattctaa taacttcatt ttttattaat tagtatttga taatataaaa tttcattagt
     2641 tgctttcaaa cattattcag acaatctgtc atatggataa aaagtatagt attggtttag
     2701 atattggtac taactctgtg ggttgggcag ttatcactga cgaatataaa gttccatcaa
     2761 agaaatttaa ggtgttaggt aacactgaca gacactcaat aaaaaagaat cttatcggtg
     2821 ctcttttgtt cgactccggt gaaactgccg aggctacacg tttaaaaaga acagcaagaa
     2881 gaagatatac ccgtagaaaa aatagaatat gttatttaca agaaatcttt tctaatgaaa
     2941 tggctaaagt tgatgattcc tttttccata gattggaaga gtcatttttg gttgaagaag
     3001 acaaaaagca tgagagacat ccaatctttg ggaatatagt tgatgaagtg gcttaccatg
     3061 aaaaatatcc taccatttat catttaagaa agaaattggt agattcaact gataaagctg
     3121 accttagatt aatctattta gcacttgccc atatgattaa atttagaggt cattttttga
     3181 ttgaaggtga tttgaaccca gataattctg acgtggataa attatttatt caattagtcc
     3241 aaacctacaa ccaattattt gaggaaaatc caattaatgc tagtggtgtc gatgccaaag
     3301 ctatattatc agccagatta tcaaaatcta gacgtttgga aaatttgatt gcccaattgc
     3361 caggagaaaa aaagaatgga ttatttggaa acttgatcgc attatcattg ggtttgacac
     3421 caaattttaa atctaatttt gatttagctg aagatgctaa attacaatta tcaaaagaca
     3481 cctatgacga cgatttggac aatttacttg ctcaaattgg tgatcaatat gcagatttgt
     3541 tcttagctgc taaaaactta tctgatgcta ttttgttgtc tgatattttg agagtgaaca
     3601 cagaaataac caaagctcca ttatcagcat ctatgatcaa acgttatgat gaacaccatc
     3661 aggatttgac tttattgaaa gctttggtga gacaacaatt gccagagaag tataaagaaa
     3721 tctttttcga tcaatctaaa aacgggtatg caggttatat tgatgggggt gcctcccaag
     3781 aggaatttta caaatttata aaacctattt tagaaaagat ggatgggact gaggaacttt
     3841 tggtcaaatt gaacagagaa gatttgttac gtaaacagag aacttttgat aatggtagta
     3901 tacctcacca aattcatttg ggtgagttgc atgcaatttt aagaagacaa gaagattttt
     3961 atccattttt aaaagataat agagaaaaaa tcgagaaaat tttaaccttt agaattccat
     4021 actatgttgg gcctttggct agaggtaatt caagatttgc ctggatgaca cgtaaatcag
     4081 aagaaactat taccccttgg aattttgaag aggttgttga taaaggagca tcagcacaga
     4141 gttttattga aagaatgacc aatttcgata aaaacttacc aaatgaaaaa gttttaccaa
     4201 aacattcctt gttatacgaa tattttactg tttacaatga acttacaaag gttaaatatg
     4261 ttactgaagg tatgcgtaag ccagcctttt tatctggaga acagaaaaag gcaatagttg
     4321 atttattgtt taaaacaaat agaaaagtta ctgttaaaca attaaaagaa gattacttta
     4381 agaaaattga atgttttgat tcagttgaaa tcagtggtgt tgaagacaga tttaatgcta
     4441 gtttaggaac ttaccatgat ttacttaaaa ttatcaaaga taaagatttc ttggataacg
     4501 aagaaaatga agacatttta gaagacattg ttttaacctt aactttattc gaagatagag
     4561 agatgattga agaacgtttg aagacttatg cacatttgtt tgacgataaa gtgatgaaac
     4621 agttgaaaag aagacgttat actggatggg gtagattgtc tcgtaaattg atcaatggaa
     4681 ttagagataa acaaagtggt aaaactatct tggacttttt gaaatctgac ggatttgcta
     4741 atagaaattt catgcaattg atccacgacg atagtttgac atttaaagaa gacatccaaa
     4801 aggcccaagt gagtgggcaa ggtgattcat tacatgaaca tattgcaaat ttagccggat
     4861 ctcctgctat taagaaaggg atattacaaa ctgttaaagt tgtggatgaa ttagtgaaag
     4921 taatgggaag acataaacct gaaaacattg tcattgagat ggcaagagaa aatcaaacta
     4981 cacaaaaagg acagaaaaat agtagagaac gtatgaaaag aatagaagag ggtattaaag
     5041 aattgggtag tcaaatattg aaagaacacc cagtggaaaa tacccagttg caaaatgaaa
     5101 aattatatct ttactacctt caaaatggac gtgatatgta tgttgatcag gaattagata
     5161 taaatagact ttcagattat gatgtagatc atatagttcc acaatctttc ttgaaagatg
     5221 attccataga caataaagta ttaactagaa gtgataaaaa tagaggtaaa agtgataatg
     5281 tcccaagtga ggaagtcgtc aaaaagatga aaaattactg gcgtcaactt ttgaatgcta
     5341 aattaattac tcaaagaaaa tttgataatt tgactaaagc agaaagaggt gggctttctg
     5401 aattagataa agccgggttc attaaaagac aattggtcga aactagacaa attactaaac
     5461 atgttgccca aattttagat tcccgtatga acactaagta tgacgaaaat gataagttaa
     5521 tacgtgaggt taaagtcatt actttaaaat caaaacttgt ctctgatttc agaaaggatt
     5581 tccaattcta taaagttaga gaaattaata attatcatca tgctcatgat gcatatttga
     5641 atgctgtagt tggaactgct ttaatcaaga aataccctaa attagaatct gaatttgtat
     5701 atggtgatta caaagtctat gatgttagaa agatgattgc taaatcagaa caagaaattg
     5761 gtaaagctac agctaaatac ttcttttact ctaacattat gaatttcttt aaaacagaaa
     5821 ttactttggc aaacggtgaa attagaaaaa gacctcttat tgaaacaaat ggtgagactg
     5881 gagagatagt ttgggacaaa gggcgtgatt tcgctactgt tagaaaagtt ttatcaatgc
     5941 cacaagttaa cattgtaaag aaaacagagg ttcaaactgg tggtttctca aaagaaagta
     6001 ttttgcctaa aagaaatagt gataaattga ttgccagaaa aaaggattgg gatccaaaga
     6061 aatatggtgg tttcgactca ccaaccgtag cctattctgt tttggttgtg gcaaaggttg
     6121 aaaagggtaa aagtaaaaag cttaaatcag taaaagaact tttgggtatt acaataatgg
     6181 aaagaagttc ctttgaaaag aaccctattg attttttgga agctaaaggt tataaggaag
     6241 taaagaagga cttaataatc aaattgccta aatattcttt atttgaatta gaaaatggga
     6301 gaaaaagaat gttggcttct gctggagaat tgcaaaaggg taatgaatta gcattgcctt
     6361 ccaaatatgt taacttcttg tatttagctt cacactatga aaagttgaaa gggtcaccag
     6421 aagataacga gcaaaaacaa ttatttgttg aacaacacaa acactactta gatgagatta
     6481 tagaacaaat tagtgaattc agtaaaagag tgatattagc tgatgcaaat ttagataaag
     6541 ttttgtcagc ctataacaaa catagagata agccaattag agaacaagca gaaaacatta
     6601 ttcacttatt tacccttacc aatttaggag cacctgctgc tttcaagtat tttgatacaa
     6661 caattgatcg taaaagatat acctcaacaa aagaagtctt agacgccacc ttaattcatc
     6721 aatcaatcac tggattgtat gagacaagaa ttgatttgtc tcaattgggt ggtgatgaag
     6781 gggctgatcc taagaagaaa agaaaagttg atccaaagaa aaagcgtaag gtggatccta
     6841 agaaaaagag aaaggttgac tacaaagacc atgacggtga ttataaagat catgacatcg
     6901 actacaagga tgacgatgac aagtgataat gactgcagag atccatcgac ctgccgccaa
     6961 gctaattccg ggcgaatttc tgtcgagtca tgtaattagt tatgtcacgc ttacattcac
     7021 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     7081 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     7141 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     7201 agaaggtttt gggacgctcg aaatcgcgtg cattcaacgc tgcaggtcga caacccttaa
     7261 tataacttgg aaagcaagca atcgtagccc cagcaaagca ctatagtagg tgtttgctct
     7321 ttttatttcc tttgctctac gcactgtaat acaggattga tgaaagcagt atagaaaatc
     7381 gcctcgaagc ctcgtgaatc agtagatcta tttacattgt tcgtctcgtg cggtttttca
     7441 ccagtgtcat gagcgagcga acttggtcga catcattaaa aaaaactgcg gctttcgtgt
     7501 cagagtcgct gtgtatcact tcgacgtagg taatcagctc ttaagcttga agtgtggatc
     7561 ggaattttct tctcctcctt cattagtcat cttccatgaa agagtgactt ttgactcaca
     7621 cgaaggaaaa tgtgcgcaaa ttacaaatca cctacaagcc acaagacaca ttcatcaaag
     7681 gcaccaaaca agacattcaa gtggaccatg ataaaagcca aggtttgaag gcatattctg
     7741 tatacttcaa tcttggtttg ctcggcccaa gtcccggcga ttagcggcgc ccggtggccc
     7801 ggctggatag atttcaattg gcacacgcac aattatctcc acaattgcat tgcatacacc
     7861 tttacaccac aactcggcga cctcggccag cctctgaaaa cacatagcca aaaacgaaga
     7921 agggacaggc attttccctt cattgcctag tatggcataa tacgtcacca aaccatgcat
     7981 tatggattgt taagagctaa tcttcctggt gaaaaaccgc cacaaattga ccatgacagt
     8041 gcttgctcgg agttttcagc ggagggcgtg cgcccgggca aacatagtgc attgtgatgt
     8101 aggcggcaaa tatatgccgg atacgcgctt atggagcgga ttgccccggc ggagaaaagt
     8161 atataagtag ccagcgcaag gcccccattt ctagagaatt tttcccttca attgattaca
     8221 acttcaatta actacaaaca aaacaaacaa ccatgggaac caccttggat gacactgctt
     8281 atcgataccg aacatctgta cctggtgatg ctgaagctat cgaagccctt gacgggtcat
     8341 tcacaactga caccgtcttt agagttacag ctactgggga tgggtttaca ttgcgagaag
     8401 taccagtaga tcctcctttg accaaagtat tcccagatga cgaaagtgac gatgaatccg
     8461 atgacggtga ggacggtgac ccagactcaa gaacatttgt cgcttatgga gacgacggtg
     8521 atttggccgg tttcgtagtt gtatcctatt ctggatggaa ccgaagactt actgtcgagg
     8581 atatagaggt tgccccagaa catcgtgggc acggtgttgg gcgagccctt atgggattag
     8641 ctacagaatt tgcccgtgaa cgtggggctg gacatttatg gttagaagtc accaatgtta
     8701 atgcaccagc tatacatgca tatcgaagaa tggggttcac actttgtgga ttggatacag
     8761 ctttatacga tggtacagcc tcagacggag aacaagcttt atacatgtct atgccttgtc
     8821 cataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt
     8881 tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg
     8941 cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgttaa
     9001 tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt
     9061 gtcgaaaacg agctctcgag aacccttaat ataacttcgt ataatgtatg ctatacgaag
     9121 ttattaggtg atatcagatc cactagtggc cattaagacc tcagcgcggc cgcaaattta
     9181 aataaaatga agtgaagttc ctatactttc tagagaatag gaacttctat agtgagtcga
     9241 ataagggcga cacaaaattt attctaaatg cataataaat actgataaca tcttatagtt
     9301 tgtattatat tttgtattat cgttgacatg tataattttg atatcaaaaa ctgattttcc
     9361 ctttattatt ttcgagattt attttcttaa ttctctttaa caaactagaa atattgtata
     9421 tacaaaaaat cataaataat agatgaatag tttaattata ggtgttcatc aatcgaaaaa
     9481 gcaacgtatc ttatttaaag tgcgttgctt ttttctcatt tataaggtta aataattctc
     9541 atatatcaag caaagtgaca ggcgccctta aatattctga caaatgctct ttccctaaac
     9601 tccccccata aaaaaacccg ccgaagcggg tttttacgtt atttgcggat taacgattac
     9661 tcgttatcag aaccgcccag ggggcccgag cttaagactg gccgtcgttt tacaacacag
     9721 aaagagtttg tagaaacgca aaaaggccat ccgtcagggg ccttctgctt agtttgatgc
     9781 ctggcagttc cctactctcg ccttccgctt cctcgctcac tgactcgctg cgctcggtcg
     9841 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     9901 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     9961 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
    10021 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
    10081 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
    10141 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
    10201 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
    10261 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
    10321 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
    10381 cagagttctt gaagtggtgg gctaactacg gctacactag aagaacagta tttggtatct
    10441 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
    10501 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
    10561 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgacg
    10621 cgcgcgtaac tcacgttaag ggattttggt catgagcttg cgccgtcccg tcaagtcagc
    10681 gtaatgctct gcttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    10741 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    10801 accatctggc cccagcgctg cgatgatacc gcgagaacca cgctcaccgg ctccggattt
    10861 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    10921 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    10981 tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    11041 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    11101 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    11161 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    11221 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    11281 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    11341 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    11401 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    11461 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    11521 aataagggcg acacggaaat gttgaatact catattcttc ctttttcaat attattgaag
    11581 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    11641 acaaataggg gtcagtgtta caaccaatta accaattctg aacattatcg cgagcccatt
    11701 tatacctgaa tatggctcat aacacccctt gtttgcctgg cggcagtagc gcggtggtcc
    11761 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtgggga
    11821 ctccccat
//
