LOCUS       sequence-208542-       10749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Escherichia coli- S. cerevisiae shuttle plasmid harbors a Cas9 gene,
            a natMX6 and URA3 selection markers used in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10749)
  AUTHORS   Wang X, Yang J, Yang S, Jiang Y
  TITLE     Unraveling the genetic basis of fast l-arabinose consumption on top 
            of recombinant xylose-fermenting Saccharomyces cerevisiae.
  JOURNAL   Biotechnol Bioeng. 2018 Sep 10. doi: 10.1002/bit.26827.
  PUBMED    30199094
REFERENCE   2  (bases 1 to 10749)
  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: "Biotechnol 
            Bioeng. 2018 Sep 10. doi: 10.1002/bit.26827."
COMMENT     Sequence Label: pHCas9-Nours
FEATURES             Location/Qualifiers
     source          1..10749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..400
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             420..440
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             441..4541
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             441..4541
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             441..4541
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4554..4574
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4590..4837
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4606..4624)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4969..4988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5059..5402
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        5403..5450
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5469..6041
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      6047..6244
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     6456..6477
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6745..7548)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(7549..7764)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7788..7807)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7907..7929
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7967..7985)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    8026..8529
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        8566..8670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8671..8739,8740..9531)
                     /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(8889..8908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9702..10290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10191..10210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10444..10461
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10578..10599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10614..10619,10620..10637,10638..10644)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10652..10668
                     /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     10657..10679
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10676..10692
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10676..10692
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10711..10731
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10713..10731
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 catagcttca aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc
       61 atcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct
      121 ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt
      181 ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa
      241 tttttttttt tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac
      301 ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact
      361 tcttgctcat tagaaagaaa gcatagcaat ctaatctaag ttttaattac aaaatgcctc
      421 caaaaaagaa gagaaaggtc gacaagaagt actccattgg gctcgatatc ggcacaaaca
      481 gcgtcggttg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
      541 tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact
      601 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
      661 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
      721 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
      781 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
      841 tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct
      901 atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga
      961 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
     1021 ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta
     1081 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
     1141 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
     1201 acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc
     1261 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
     1321 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
     1381 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     1441 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     1501 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     1561 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca
     1621 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     1681 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     1741 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     1801 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
     1861 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     1921 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     1981 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     2041 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     2101 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
     2161 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     2221 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     2281 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     2341 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc
     2401 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga
     2461 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     2521 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     2581 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     2641 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     2701 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     2761 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     2821 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     2881 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg
     2941 actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata
     3001 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     3061 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     3121 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     3181 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     3241 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     3301 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
     3361 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     3421 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag
     3481 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     3541 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     3601 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     3661 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg
     3721 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     3781 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     3841 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     3901 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg
     3961 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca
     4021 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     4081 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     4141 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
     4201 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     4261 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     4321 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
     4381 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
     4441 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     4501 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga
     4561 agaagaggaa ggtgtgatct cttctcgagt catgtaatta gttatgtcac gcttacattc
     4621 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     4681 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     4741 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     4801 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtaccttc ccaacagttg
     4861 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4921 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4981 tcttcccttc ctttctcgcc acgttcgccc tagagatctg tttagcttgc ctcgtccccg
     5041 ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg
     5101 cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta
     5161 taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc
     5221 tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac
     5281 gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat
     5341 atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa
     5401 ccgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     5461 actaaaccat gggtaccact cttgacgaca cggcttaccg gtaccgcacc agtgtcccgg
     5521 gggacgccga ggccatcgag gcactggatg ggtccttcac caccgacacc gtcttccgcg
     5581 tcaccgccac cggggacggc ttcaccctgc gggaggtgcc ggtggacccg cccctgacca
     5641 aggtgttccc cgacgacgaa tcggacgacg aatcggacga cggggaggac ggcgacccgg
     5701 actcccggac gttcgtcgcg tacggggacg acggcgacct ggcgggcttc gtggtcgtct
     5761 cgtactccgg ctggaaccgc cggctgaccg tcgaggacat cgaggtcgcc ccggagcacc
     5821 gggggcacgg ggtcgggcgc gcgttgatgg ggctcgcgac ggagttcgcc cgcgagcggg
     5881 gcgccgggca cctctggctg gaggtcacca acgtcaacgc accggcgatc cacgcgtacc
     5941 ggcggatggg gttcaccctc tgcggcctgg acaccgccct gtacgacggc accgcctcgg
     6001 acggcgagca ggcgctctac atgagcatgc cctgccccta atcagtactg acaataaaaa
     6061 gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt
     6121 ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc
     6181 gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat
     6241 actgctgtcg attcgatact aacgccgcca tccagtgtcg aaaacgggct ttccccgtca
     6301 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     6361 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6421 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6481 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     6541 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     6601 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     6661 cgcatagggt aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca
     6721 tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc
     6781 ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag
     6841 tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata
     6901 ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca
     6961 atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc
     7021 tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc
     7081 cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc
     7141 cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc
     7201 tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat
     7261 gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag
     7321 cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg tttttagtaa
     7381 acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc
     7441 caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac
     7501 aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg
     7561 tttcctgcag gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc
     7621 ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc
     7681 cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga
     7741 ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca ctctcagtac aatctgctct
     7801 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     7861 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     7921 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     7981 ctatttttat aggttaatgt catgataata atggtttctt agacggatcg cttgcctgta
     8041 acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta
     8101 tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta gaagagttac
     8161 ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag cgtactttac
     8221 atatatattt attagacaag aaaagcagat taaatagata tacattcgat taacgataag
     8281 taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag atgaaacaat
     8341 tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg ttggcgatcc
     8401 ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat ttcttttttt
     8461 actttctatt tttaatttat atatttatat taaaaaattt aaattataat tatttttata
     8521 gcacgtgatg aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     8581 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     8641 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     8701 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     8761 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     8821 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     8881 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     8941 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     9001 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     9061 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     9121 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     9181 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     9241 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     9301 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     9361 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     9421 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     9481 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     9541 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     9601 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     9661 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     9721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     9781 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     9841 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     9901 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     9961 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
    10021 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
    10081 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
    10141 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
    10201 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
    10261 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
    10321 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
    10381 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
    10441 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
    10501 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
    10561 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
    10621 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
    10681 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
    10741 ctggagctc
//
