LOCUS       sequence-198860-       11362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for S. pombe CRISPR-Cas9 system. The gRNA can be 
            integrated easily through Gibson Assembly with the plasmid backbone 
            digested by Not1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11362)
  AUTHORS   Zhao Y, Boeke JD
  TITLE     Construction of Designer Selectable Marker Deletions with 
            CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized 
            Design of Common Entry Vectors.
  JOURNAL   G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 
            10.1534/g3.117.300363.
  PUBMED    29321167
REFERENCE   2  (bases 1 to 11362)
  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: "G3 
            (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 
            10.1534/g3.117.300363."
COMMENT     Sequence Label: pYZ033
FEATURES             Location/Qualifiers
     source          1..11362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(75..322)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     288..306
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(338..358)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(371..4474)
                     /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             complement(371..4474)
                     /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             complement(371..4474)
                     /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"
     promoter        complement(4495..5236)
                     /label=adh1 promoter
                     /note="promoter for the Schizosaccharomyces pombe adh1 gene
                     encoding alcohol dehydrogenase"
     promoter        5265..5369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5370..5438,5439..6230)
                     /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(5588..5607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6401..6989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6890..6909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7143..7160
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7277..7298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7313..7318,7319..7336,7337..7343)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7351..7367
                     /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     7356..7378
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7375..7391
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7375..7391
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7428..7784
                     /label=rrk1 promoter/leader
                     /note="promoter and leader RNA for the Schizosaccharomyces 
                     pombe rrk1 gene encoding the K-RNA component of RNase P 
                     (Jacobs et al., 2014)"
     misc_RNA        7877..7928
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     primer_bind     complement(7978..7995)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7978..7994)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7987..8009)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(8635..9429)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     rep_origin      10039..10822
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(11193..11212)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11312..11334
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacggg gatcgcaaat taaagccttc gagcgtccca aaaccttctc aagcaaggtt
      121 ttcagtataa tgttacatgc gtacacgcgt ctgtacagaa aaaaaagaaa aatttgaaat
      181 ataaataacg ttcttaatac taacataact ataaaaaaat aaatagggac ctagacttca
      241 ggttgtctaa ctccttcctt ttcggttaga gcggatgtgg ggggagggcg tgaatgtaag
      301 cgtgacataa ctaattacat gactcgagaa gagatcacac cttcctcttc ttcttggggt
      361 cagccctgct gtctccaccg agctgagaga ggtcgattct tgtttcatag agccccgtaa
      421 ttgactgatg aatcagtgtg gcgtccagga cctcctttgt agaggtgtac cgctttctgt
      481 ctatggtggt gtcgaagtac ttgaaggctg caggcgcgcc caagttggtc agagtaaaca
      541 agtggataat gttttctgcc tgctccctga tgggcttatc cctgtgctta ttgtaagcag
      601 aaagcacctt atcgaggtta gcgtcggcga ggatcactct tttggagaat tcgcttattt
      661 gctcgatgat ctcatcaagg tagtgtttgt gttgttccac gaacagctgc ttctgctcat
      721 tatcttcggg agaccctttg agcttttcat agtggctggc cagatacaag aaattaacgt
      781 atttagaggg cagtgccagc tcgttacctt tctgcagctc gcccgcacta gcgagcattc
      841 gtttccggcc gttttcaagc tcaaagagag agtacttggg aagcttaatg atgaggtctt
      901 ttttgacctc tttatatcct ttcgcctcga gaaagtcgat ggggtttttt tcgaagcttg
      961 atcgctccat gattgtgatg cccagcagtt ccttgacgct tttgagtttt ttagacttcc
     1021 ctttctccac tttggccaca accagtacac tgtaagcgac tgtaggagaa tcgaatccgc
     1081 cgtatttctt ggggtcccaa tcttttttgc gtgcgatcag cttgtcgctg ttccttttcg
     1141 ggaggatact ttccttggag aagcctccgg tctgtacttc ggtcttttta acgatgttca
     1201 cctgcggcat ggacaggacc ttccggactg tcgcgaaatc cctacccttg tcccacacga
     1261 tttctcctgt ttctccgttt gtttcgataa gtggtcgctt ccgaatctct ccattggcca
     1321 gtgtaatctc ggtcttgaaa aaattcataa tattgctgta aaagaagtac ttagcggtgg
     1381 ccttgcctat ttcctgctca gactttgcga tcattttcct aacatcgtac actttatagt
     1441 ctccgtaaac aaattcagat tcaagcttgg gatatttttt gataagtgca gtgcctacca
     1501 ctgcattcag gtaggcatca tgcgcatggt ggtaattgtt gatctctctc accttataaa
     1561 actgaaagtc ctttctgaaa tctgagacca gcttagactt cagagtaata actttcacct
     1621 ctcgaatcag tttgtcattt tcatcgtact tggtgttcat gcgtgaatcg agaatttggg
     1681 ccacgtgctt ggtgatctgg cgtgtctcaa caagctgcct tttgatgaag ccggctttat
     1741 ccaactcaga caggccacct cgttcagcct tagtcagatt atcgaacttc cgttgtgtga
     1801 tcagtttggc gttcagcagc tgccgccaat aatttttcat tttcttgaca acttcttctg
     1861 aggggacgtt atcactcttc cctctatttt tatcggatct tgtcaacact ttattatcaa
     1921 tagaatcatc tttgagaaaa gactggggca cgatatgatc cacgtcgtag tcggagagcc
     1981 gattgatgtc cagttcctga tccacgtaca tgtccctgcc gttctgcagg tagtacaggt
     2041 agagcttctc attctgaagc tgggtgtttt caactgggtg ttccttaagg atttgggacc
     2101 ccagttcttt tataccctct tcaatcctct tcatcctttc cctactgttc ttctgtccct
     2161 tctgggtagt ttggttctct cgggccatct cgataacgat attctcgggc ttatgccttc
     2221 ccattacttt gacgagttca tccacgacct taacggtctg cagtattccc tttttgatag
     2281 ctgggctacc tgcaagatta gcgatgtgct cgtgaagact gtccccctgg ccagaaactt
     2341 gtgctttctg gatgtcctcc ttaaaggtga gagagtcatc atggatcaac tgcatgaagt
     2401 tccggttggc aaatccatcg gacttaagaa aatccaggat tgtctttcca gactgcttat
     2461 ctcggatccc attgatcagt tttcttgaca gccgccccca tcctgtatat cggcgcctct
     2521 tgagctgttt catgactttg tcgtcgaaga gatgagcgta agttttcaag cgttcttcaa
     2581 tcatctccct atcttcaaac aacgtaaggg tgaggacaat gtcctcaaga atgtcctcgt
     2641 tctcctcatt gtccaggaag tccttgtctt taatgatttt caggagatcg tgatacgttc
     2701 ccagggatgc gttgaagcga tcctccactc cgctgatttc aacagagtcg aaacattcaa
     2761 tctttttgaa atagtcttct ttgagctgtt tcacggtaac tttccggttc gtcttgaaga
     2821 ggaggtccac gatagctttc ttctgctctc cagacaggaa tgctggcttt ctcatccctt
     2881 ctgtgacgta tttgaccttg gtgagctcgt tataaactgt gaagtactcg tacagcagag
     2941 agtgtttagg aagcaccttt tcgttaggca gatttttatc aaagttagtc atcctttcga
     3001 tgaaggactg ggcagaggcc cccttatcca cgacttcctc gaagttccag ggagtgatgg
     3061 tctcttctga tttgcgagtc atccacgcga atctggaatt tccccgggcg agggggccta
     3121 catagtaggg tatccgaaat gtgaggattt tctcaatctt ttccctgtta tctttcaaaa
     3181 aggggtagaa atcctcttgc cgcctgagga tagcgtgcag ttcgcccagg tgaatctggt
     3241 gggggatgct tccattgtcg aaagtgcgct gtttgcgcaa cagatcttct ctgttaagct
     3301 ttaccagcag ctcctcggtg ccgtccattt tttccaagat gggcttaata aatttgtaaa
     3361 attcctcctg gcttgctccg ccgtcaatgt atccggcgta gccattttta gactgatcga
     3421 agaaaatttc cttgtacttc tcaggcagtt gctgtctgac aagggccttc agcaaagtca
     3481 agtcttggtg gtgctcatca tagcgcttga tcatactagc gctcagcgga gctttggtga
     3541 tctccgtgtt cactcgcaga atatcactca gcagaatggc gtctgacagg ttctttgccg
     3601 ccaaaaaaag gtctgcgtac tggtcgccga tctgggccag cagattgtcg agatcatcat
     3661 cgtaggtgtc tttgctcagt tgaagcttgg catcttcggc caggtcgaag ttagatttaa
     3721 agttgggggt cagcccgagt gacagggcga taagattacc aaacaggccg ttcttcttct
     3781 ccccagggag ctgtgcgatg aggttttcga gccgccggga tttggacagc ctagcgctca
     3841 ggattgcttt ggcgtcaact ccggatgcgt tgatcgggtt ctcttcgaaa agctgattgt
     3901 aagtctgaac cagttggata aagagtttgt cgacatcgct gttgtctggg ttcaggtccc
     3961 cctcgatgag gaagtgtccc cgaaatttga tcatatgcgc cagcgcgaga tagatcaacc
     4021 gcaagtcagc cttatcagta ctgtctacaa gcttcttcct cagatgatat atggttgggt
     4081 acttttcatg gtacgccacc tcgtccacga tattgccaaa gattgggtgg cgctcgtgct
     4141 ttttatcctc ctccaccaaa aaggactcct ccagcctatg gaagaaagag tcatccacct
     4201 tagccatctc attactaaag atctcctgca ggtagcagat ccgattcttt ctgcgggtat
     4261 atctgcgccg tgctgttctt ttgagccgcg tggcttcggc cgtctccccg gagtcgaaca
     4321 ggagggcgcc aatgaggttc ttctttatgc tgtggcgatc ggtattgccc agaactttga
     4381 attttttgct cggcaccttg tactcgtccg taatgacggc ccaaccgacg ctgtttgtgc
     4441 cgatatcgag cccaatggag tacttcttgt ccatctctag agtcgacctg cagcaattct
     4501 cttgcttaaa gaaaagcgaa ggcacctgtc caccacccac tatttatacc atgggaggcg
     4561 tcaaaccgag tttttgcgaa aggattatcg cacctctccc caagtaacat cggcaaagca
     4621 aaaacggagc ggaagaccaa tgagacgcgg aaggggagcg ccgaatgagg aaagcgagag
     4681 agaaagaagt ggatggaatg attccccgcc aaagtggaca catttcggga atcttcagga
     4741 agcatggatc accttatgga caagaagata atccggaagc caataggagg gcgacagggg
     4801 gtcagcgata agcgtcggaa aaaactttat gatatggtaa atacgtagga ggttacggtg
     4861 cagaggaggg gccggtagcg agtgatagcg agggaaagac gatcgctatc gggccgtaag
     4921 gaggaaatgg gaggttgggg gaaggatgcg ttggaatgcg gagtagagaa attgaagaga
     4981 gagagaggac acacctagag agaaagaaat ggatatagag aaagagaaaa aataagagga
     5041 atacggatac gatggagtgc gttgaatgag agaaaagaaa agaaaaggaa tgataagaga
     5101 aggaatgaaa attgttttgt tttgttttca attgctttct tttgttttct ttgtctttgt
     5161 tcttccttcc cttgtcccga tgctgttctt ctcaccagct tccgcatgat agccattttc
     5221 ttagttgttg tagggcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5281 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5341 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5401 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5461 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5521 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5581 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5641 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5701 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5761 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5821 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5881 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5941 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6001 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6061 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6121 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6181 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6241 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6301 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6361 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6421 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6481 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6541 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6601 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6661 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6721 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6781 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6841 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6901 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6961 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7021 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7081 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7141 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     7201 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     7261 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     7321 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     7381 acagctatga catgattacg aattcgagct cggtaccccg ggtacctttt gcttatgttg
     7441 gtggtagttg gcatgcgtag actgatgact agtcagcaag gagcgtagaa cagtcacact
     7501 cgttatatat gtgcttccaa gaaaactcaa gaatttacca ttagcaaaca cttttttgaa
     7561 atgttagaca tttaaatgac gaaggcatat agaagctttg aataggtgtt gtaaagtgtt
     7621 gatttatgtg acgctgaggg tgcgcatgaa aggaatgttg ggtcacgatt attaaacagt
     7681 ttgctagctt ggacacttga gtattggaag ttgttgaatt ctaaaaaact ttcagttgat
     7741 ttgaatagtt gctgttgcca aaaaacataa cctgtaccga agaagcggcc gcgttttaga
     7801 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa agtgagtggc
     7861 accgagtcgg tggtgccctg tcaccggatg tgctttccgg tctgatgagt ccgtgaggac
     7921 gaaacagggg taccgtgggg atcctctaga gtcgacctgc aggcatgcaa gcttggcact
     7981 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     8041 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     8101 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     8161 gcatctgtgc ggtatttcac accgcatacc atcgcgatga gcttgtgata ttgacgaaac
     8221 tttttgacat ctaatttatt ctgttccaac accaatgttt ataaccaagt tttatcttgt
     8281 ttgtctacat ggtattttac attcatctac atacatcttt cattggcttt gtacatagtt
     8341 atcattacaa gtctaaaaaa attcactctt ttcttattca atgtcaatcc aagagaaaag
     8401 attgtggtaa tgttgtagga gcatgtttaa taaattacta tagcaaatta ctttttattc
     8461 ccaaggtgtt tatctataat agttaatatt ttagtcgcta cataaaattt taccaaagag
     8521 tacttgtata ctaattctaa atgccttctg acataaaacg cctaggaaaa caaacgcaaa
     8581 caaggcatcg actttttcaa taaccaacca aaaaaatttt acattagtct ttttttaatg
     8641 ctgagaaagt ctttgctgat atgccttcca accagcttct ctatatctct tggcttcgac
     8701 aacaggatta cgaccagctc catagactcc acgaccaaca atgatgatat cgctaccgca
     8761 gtttacaatc acttcttcag gagtacgata ttgctgtccc agcccgtctc ctttaacatc
     8821 caagccgata ccaggggaca tagttatgta gtcgctttga aggttaggaa atcgacgacc
     8881 agctataaag ccaaagcaaa aatcggtatg cttctcaaac cattctaagg ttttctctgt
     8941 gtaggaacca gtagccaaag agcctttgga agacatttca gccaaaagca agagaccacg
     9001 tcccaaaggt aaaccaactt ctttgaggcc ttgtataata ccctcgcctg gcactgtatg
     9061 gcaatttgtg atatgagccc aagaagcaat tttgtacaca ccagatgcat attgtagctt
     9121 gacggtattt ccaatgtctg cgaatttgcg atcctcaaag ataagaaaac gatgcttttt
     9181 acctaaggcc accagttttt ctaccatatc ctggtcgaaa tcctcgacaa cgtcaatatg
     9241 tgtcttgata acacagacat agggtccaat tttatctacc aattctaaga tttcggattt
     9301 cttcgtcaaa tcgaccgcga ctgacaagtt gctttgcttt tcttccatca aagccaacaa
     9361 ttccttggca atgggatttt tcatcccctc agctctagct gaatagcttt gaaatactct
     9421 agcatccata actttgcttt taaaccttta atttcgatcc aagcaaaaaa gaggttcttg
     9481 gtaggacaat acggtaagaa aacacgacat gtgcagagat gccgacgaag catagttaaa
     9541 ctgggatggt aaaatcaatt aagaatttat aaagacaaaa ttgtataagt ctctaaaaca
     9601 tcttaattat acctcacaga actatctaaa atatattcac aaagtgcaaa cattatcatg
     9661 aaaaagaacc attttaattt aaagcaaggg cattaaggct tatttacaga atttcttact
     9721 tttgtaaaga ttataaggct gattatcttt ttcaccatgc caaaaattac acaagataga
     9781 atggatgttt gaaattaaac gtgagtatac aaacaaatac actaggtaaa tcgaaacatt
     9841 tttttctcca ttaagtaaca aattcctatt tagagaaaga atgctgagta gattaaataa
     9901 tctatacaaa cttttttaac acaaatgcat acatatagcc agtgggattt gtagctaagc
     9961 tcatcgatgg cgagtctaac tccttaacca ctcggacata gtgacttatc tgacactcat
    10021 tgtaaattaa tatatatata ggcattttgt ttagttaaag gtacttaagt aattagtata
    10081 aacgaaccaa ttttataatc aggaagttaa gtgaatggta gcacatgtcg taaaaattgt
    10141 gaatttttat tgaataatat tttaaataca agcctttcta gactaggtat actcataaac
    10201 nnatatgagc aaaaggatag aggagattac attgcatctt ctacaaattt atttattgcc
    10261 ctttactgaa aaattaaata atgagtacta aatgataaaa agcgctcagt acaagaaaga
    10321 ttgcaaaaat attgcattct tcatgaatta aagttgcata taaggcatat tgaaagtaat
    10381 agtactaaaa cagcagttag cgaaaattaa tagaattata ttcgcaagac aattgtgaca
    10441 ttaaattaaa aaattgtaga atttttacta tcctctttaa cgccatgagc ctttataaaa
    10501 aggttaaatt agttttaaca ttcttttttt gagtaagagt ttacaattta tcaaaacctg
    10561 tgttattata ttcattagtt tcaatttatt agcatctaga gaaaaatcaa ttggcagtta
    10621 cattgttgga atttatgaag aaaaagaatc tacaacggag aataagttgc tgatcgcttt
    10681 ccctaaaatt gtatattttg ctgagcttat tttgacattt cgttgaagtt ttctctaatt
    10741 cgcattcatt ttaagtaaaa caatgagaaa taaaattaca aaaaatacaa attaaaatac
    10801 aatttttagc tataattata gacgatgccc ttgtatccca ttctgtctcg cttgccccta
    10861 ctttttatct tttatatacc ataatgaacg ctgccgctac taaccatacc ccgattttac
    10921 atttcggact cccaaggacg tacaaaatag aaaactatag aaaaaaataa tcagaaaata
    10981 gcatgtcatc tctttgtaaa acgcgtttgc aagaagaaag gaaacaatgg agaagagatc
    11041 atccatttgt atgtaaattt tagtaaactt gaagaaatca ctaacaactt ctcttactta
    11101 gggattctat gcaaaacctt gtaaatcatc tgatggagga ctcgatttaa tgaattggaa
    11161 ggttggccat cgatggtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
    11221 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
    11281 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
    11341 ccgtcatcac cgaaacgcgc ga
//
