LOCUS       sequence-254525-       12478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon optimised Streptococcus pyogenes CAS9 ORF regulated by the 
            Cryptococcus neoformans TEF1 promoter and terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12478)
  AUTHORS   Arras SD, Chua SM, Wizrah MS, Faint JA, Yap AS, Fraser JA
  TITLE     Targeted Genome Editing via CRISPR in the Pathogen Cryptococcus 
            neoformans.
  JOURNAL   PLoS One. 2016 Oct 6;11(10):e0164322. doi: 
            10.1371/journal.pone.0164322. eCollection 2016.
  PUBMED    27711143
REFERENCE   2  (bases 1 to 12478)
  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: "PLoS One.";
            date: "2016-10-6"; volume: "11(10):e0164322. doi"; pages: " 
            10.1371/journal.pone.0164322. eCollection 2016"
COMMENT     Sequence Label: pSDMA65
FEATURES             Location/Qualifiers
     source          1..12478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        801..905
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(906..974,975..1766)
                     /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     complement(1124..1143)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1937..2525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2426..2445
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2679..2696
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2813..2834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2849..2854,2855..2872,2873..2879)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2887..2903
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2892..2914
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2911..2927
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2911..2927
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2946..2966
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2948..2966
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             4225..8328
                     /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             4225..8328
                     /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             4225..8328
                     /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             8341..8361
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(9251..9270)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9252..9270)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9280..9297)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9280..9296)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9289..9311)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             10407..11432
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             10407..11432
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
ORIGIN
        1 acacgatgtt ttccatcagt aacatcgggg cacttgtaca tattctccca ggcagacaaa
       61 gatatgacaa gaccttcacg gccgacggtt gcaacatcct tctccaattt gagctgctac
      121 aatctcctgc ttcatggcat tccagtcaca tcaaggtatc catctctttc ggcatagcta
      181 atctaggaag aatgaatata ccacgtctat aactgatcag acctaaaggg cttcagtatc
      241 ggttatacac caggatcaat ggcaaataag actcacaaat tgggatgatc atcagcagat
      301 ggtgttattc aacgggcaga ccgtgagtac aaagtgttca gcgttgaatg gcaccgtata
      361 tacaaatatg ctgagcctgt tataacctta gcttcttctc ctgttgcttc ccttggcctg
      421 taaagtcaag ttccgccgtg ttctattatc ataaatgatc gacttccatc aatggaaggg
      481 taagctcgcg tccttaagca taggcggtct cagagtgcca tccagtccag cgctcaaggt
      541 tcctaggaag gggggttcgc ctttaagaat tcagggggct gtggtagctc gacgatgcca
      601 ggtgtcatgg agaggtccaa gaaattgatg ataaccattc cctagtccgc aaagtgcctc
      661 gtatgatcca aagtgccaag attgtacgat agggtggata tccaaagcaa gaagataaac
      721 gtccaattcc atgatttgaa gtctatgggc agatcttgcc gctacagggc gcgtcaggtg
      781 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      841 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      901 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      961 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1021 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1081 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1141 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1201 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1261 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1321 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1381 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1441 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1501 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1561 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1621 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1681 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1741 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1801 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1861 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1921 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1981 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2041 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2101 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2161 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2221 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2281 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2341 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2401 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2461 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2521 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2581 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2641 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2701 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2761 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2821 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2881 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2941 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc
     3001 gccattctat gctatacggt acaagattgt ggctactatg caagctccat agcgaatgat
     3061 cttacactga ccttgtcgac cgttctgaat gattaacaat ggtcgcaagt gtggttgacc
     3121 cacctgcggg cgcttcttaa ctatctatca tcggtatgtc aagtatcaca ccctttaaac
     3181 aaaagaataa gagcataata cagcagtctc atcatggcaa cccttcgcgc aatgtacaaa
     3241 aagcagccag aagcccccac gcacaagcaa taatgtcgtg ttctaccctg ccatgaacaa
     3301 gcgtcatggc cattagcgaa cgccccaata ttcagcccca ttcgccaagc aagttgcctc
     3361 gcccccttcc cagaagatca tgggcatctc gtcttcttct aatttcgcat tttgtgaagc
     3421 tccattcatg cacgtcggct tctggacgac tcttatcata tgcattggcc tgctcggtgt
     3481 gaggatgagc cactcccaca tgtttagacg gcaggggcct cttgccaccc gttgaatttt
     3541 tggtgcgaag acgtgcctgg gaacgacctc cactttaaat ttctccgttg ttgcggtcgc
     3601 tccatcctcc tcacatttct tgcctcctaa gtcccatgtg ccacactcgg tgaaaaacgg
     3661 agtttgatac ggatgaaatt tccagtgagc ttttcacatt ctggaggttt cgaaaaaaat
     3721 tgtgggattt tctgaattaa tccgagaatc cgccctgccc atcaaccgga aagaattaaa
     3781 tccctcgcac attcctttct cccaccacct tctttatctc tacattcttt aaccaaaaca
     3841 cctaggtgag tacccaaagt cttttgacac ctttgaaaat ccgcgccagt cgtagggcac
     3901 gtaaaacaat tctcgctggt ggcttctgca ggtgaagtcg gggcagagtg gagctgagca
     3961 tggtcgacca cccacatcat tctgtcttgt ttttggcgat gtcttccatg cttttgccct
     4021 catctgttgc gtcgactgca gagtactgtt ttgccactac tgagctcaac atgggcttgt
     4081 gtgtggtgaa gaaaaaaatt tggtggctct tgaaaatttt caacccatcg gatacatgcc
     4141 cttccaactc acaacgccct aacactttat tgtcctatca ttgcctcttc gcacatgcta
     4201 acgatctcca cagaaaactt caaaatggac aagaagtaca gcattgggct cgatatcggc
     4261 acaaacagcg tcggttgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc
     4321 aaagttctgg gcaataccga tcgccacagc ataaagaaga acctcattgg cgccctcctg
     4381 ttcgactccg gggagacggc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat
     4441 acccgcagaa agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag
     4501 gtggatgact ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag
     4561 cacgagcgcc acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac
     4621 ccaaccatat atcatctgag gaagaagctt gtagacagta ctgataaggc tgacttgcgg
     4681 ttgatctatc tcgcgctggc gcatatgatc aaatttcggg gacacttcct catcgagggg
     4741 gacctgaacc cagacaacag cgatgtcgac aaactcttta tccaactggt tcagacttac
     4801 aatcagcttt tcgaagagaa cccgatcaac gcatccggag ttgacgccaa agcaatcctg
     4861 agcgctaggc tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag
     4921 aagaagaacg gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt
     4981 aaatctaact tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga cacctacgat
     5041 gatgatctcg acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg
     5101 gcaaagaacc tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc
     5161 accaaagctc cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg
     5221 actttgctga aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc
     5281 gatcagtcta aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt
     5341 tacaaattta ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag
     5401 cttaacagag aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac
     5461 cagattcacc tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt
     5521 ttgaaagata acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta
     5581 ggccccctcg cccggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc
     5641 atcactccct ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc
     5701 gaaaggatga ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct
     5761 ctgctgtacg agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa
     5821 gggatgagaa agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc
     5881 ttcaagacga accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt
     5941 gaatgtttcg actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga
     6001 acgtatcacg atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac
     6061 gaggacattc ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt
     6121 gaagaacgct tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag
     6181 aggcgccgat atacaggatg ggggcggctg tcaagaaaac tgatcaatgg gatccgagac
     6241 aagcagagtg gaaagacaat cctggatttt cttaagtccg atggatttgc caaccggaac
     6301 ttcatgcagt tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa
     6361 gtttctggcc agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct
     6421 atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga
     6481 aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag
     6541 ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg
     6601 tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac
     6661 ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaatcgg
     6721 ctctccgact acgacgtgga tcatatcgtg ccccagtctt ttctcaaaga tgattctatt
     6781 gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa cgtcccctca
     6841 gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc
     6901 acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat
     6961 aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc
     7021 caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag
     7081 gtgaaagtta ttactctgaa gtctaagctg gtctcagatt tcagaaagga ctttcagttt
     7141 tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg
     7201 gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt ttacggagac
     7261 tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc
     7321 accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg
     7381 gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc
     7441 gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg
     7501 aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg
     7561 aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc
     7621 ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg
     7681 aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca
     7741 agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa
     7801 gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga
     7861 atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac
     7921 gttaatttct tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat
     7981 gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa
     8041 ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct
     8101 gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat tatccacttg
     8161 tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac
     8221 agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt
     8281 acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag cagggctgac
     8341 cccaagaaga agaggaaggt gtgattactt atgtagtgac ccttcttacg cgtaatatgc
     8401 atcggatgac tgttgctact tgtgcttctt acgttttgtg tcttatttgc tgatgtatac
     8461 ctctgcgctt caaaatcttc attgccgtga tatatcattc catgtcgttt gtccgttatt
     8521 ggattcccga tagttgtccg cgaaatcgaa tatttctcgg ctgacgacgt ctcttgcaac
     8581 aataaaatgg ccatcactgt tgaaatttgc acgagagcta taacttggaa taaccttcgt
     8641 tgctgcttta ttttagtgct ttcaactatt tactcattct caggtcgcca ccatcgcatt
     8701 gatatcgttc cttccatcct tcctttcctt tagtcggtca cgtcttcctt aatccgtcac
     8761 tccttttttc agtcgttcct ggagatcacc acgaaataat ctcgacatgc actgaccatc
     8821 tcggttctgt ggactgtcat gctaagagag ttcatgttgt aataatagtt tgttgcatgg
     8881 aaagaacggg gaaagaagca agaaacagat gtacagaatt accgatgtct ctacgcatcc
     8941 ctgacctcgt ccttcctacc gcaaccgcac ttccacctgc tcacacgtcc tcgttaatgc
     9001 cttgaatctc agccacacac ttacctcgcc tcgacgcttg tcgggccagg gcaccccctt
     9061 catcaacctg ggaagagttg gcggggcaaa cggaggaggg atggggcagg aagaggggga
     9121 tggcgcacgt cgagtggatt tggaagaaaa ggggttggtc gaggtggaag tcgaagcgaa
     9181 gatgttgaat gtcatgtttg ggaacggggg atctttagaa gactatgaga ttgaacgggt
     9241 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     9301 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     9361 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     9421 cctgaatggc gaatggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     9481 tgttaaatca gctcattttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     9541 aaaagaatag accgagatag ggttgagtcc atggctgcga ggatgtgagc tggagagcgg
     9601 cgcacgggaa ggggaggact cacataagca tgcaggattc gagtggcatg gtgtgcactg
     9661 agtgtatggt tgtcggagga gaggatgatg gtaacaacaa tagcagcaac gtcactcgac
     9721 gcgcgtccgg tgtgccacac ggggtaacgc cgagtcgccg tcagggtcgc cgagaccact
     9781 ctcacagcgt caccgttggc accagctcag cttacagctt ctatcctccg ccagcatcca
     9841 catacatccc ctataccgca tcccccaccc actgcccaag gtgagtcatc ttcccgcccc
     9901 cttcccttgc ccgccactca gtcctccatc ctccactaat ccaccttatc gcacccaccg
     9961 cctatcgcac atccgagcac aatgctgggc ctgccagggg ctgctagatg gtgctctccc
    10021 cacgctgatc tgcatgccgg ccattggatc atgggtgcta ggtgctgggt gctggatgtt
    10081 ggatgctgga tgctgggtgc acgcttggtc atttccttcc aggattgacg gtcgccgaga
    10141 ggacgacgtg gcgttcgaca acgagggccg atagcaccgc atcgcctcga cctgcatcca
    10201 tctcgccttg tccttttggt gcaacaatcc atccgtgctg gtgccacacg catagctgga
    10261 agagatggat gtgcgttgaa cagagctgcc gtcaggactt tttggtgcac ggaccctatt
    10321 gtcctcccca atcttcaccg cgtctcctaa tatgcagcct ctttgctaat tgtctttttc
    10381 cattagtaaa ctcgcccaac atgtctatga aaaagcctga actcaccgcg acgtctgtcg
    10441 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
    10501 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
    10561 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
    10621 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
    10681 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
    10741 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
    10801 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
    10861 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
    10921 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
    10981 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
    11041 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
    11101 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
    11161 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
    11221 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
    11281 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
    11341 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
    11401 gccccagcac tcgtccgagg gcaaaggaat aggtgaaggc ggtaaggggt taattttcct
    11461 tagagggtgt atatatacat attagagaag tgatacaatt ttagcacact gcgaattcga
    11521 gacagacatc gtgtcaatca tcttttttga cattatatgc ccattaatct atctacagac
    11581 aacaatacca tccttcccac cctcagcaac gccgttgaat cctcaggatc ttcatggctc
    11641 cttgtctctg aaaccaggaa gctagtttct acatctcttc cgtgttaata cagataaacc
    11701 ggccaatgaa gcacgctcaa aattatgcct gtcaaaacag tcagctccat ccttttcttc
    11761 cctttacgta tcatatgctg tttttttttt tttttcgaca caagttcaaa gggctggcgc
    11821 tgcttaggat ccatcacacc attcgtcaac gtattgtgaa tcaaagaaat gctcttcgat
    11881 attctttatc agggcaaggg aacagacaga taccagcagg cttcacaact ctgagtcgct
    11941 cggccaaaga ggaaaagaaa agaggatgcc ggcactttct atacagtcac aagacgcgaa
    12001 tgacaatccg gcactttatc gcatcatgtc tgaatttgtg aagggacttt gagatataat
    12061 cagggggtca gacgcacttt acaactctga atcgcttcgg agtaaggaag tggaactgac
    12121 agatacactt tatagagcct tgggaaccat ctggaccaag aagaagggaa aaaaaggcta
    12181 cagtgagtga gcaacctatg ctcaccttta tcctgacgtt gattgagtct agaggccaaa
    12241 gatccaaggc aaggcgaaat attccattgt ccagttcatg gctgagtgac cgttgtttag
    12301 gttgatcgag gcgaaaagag aagtcgatgt tgatgattgt gagaccgaaa tcagacttgg
    12361 ccgcttttta tagttcaaaa aaggggccca gaggtcagtg ggctcgccac tcgatgcata
    12421 agcaatccac ttggcacaga ctggtttggt ttaattaaca caagtatcgt ggcgcgcc
//
