LOCUS       sequence-324492-       11220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/RNA scaffold based transcription regulation in Pichia 
            pastoris 
            BB3rN_pTEF2_dCAS9(3xFLAG_SV40NLS)_pPOR1_MS2bind_VP64_SV40NLS_pALD4_T
            HI11_T5_gRNA_MS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11220)
  AUTHORS   Baumschabl M, Prielhofer R, Mattanovich D, Steiger MG
  TITLE     Fine-Tuning of Transcription in Pichia pastoris Using dCas9 and RNA 
            Scaffolds.
  JOURNAL   ACS Synth Biol. 2020 Dec 18;9(12):3202-3209. doi: 
            10.1021/acssynbio.0c00214. Epub 2020 Nov 12.
  PUBMED    33180466
REFERENCE   2  (bases 1 to 11220)
  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"; doi: 
            "10.1021/acssynbio.0c00214"; journalName: "ACS Synth Biol"; date: 
            "2020-12-18- 18"; volume: "9"; issue: "12"; pages: "3202-3209"
COMMENT     Sequence Label: THI VP64 pALD4 T5
FEATURES             Location/Qualifiers
     source          1..11220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             646..711
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(715..741,742..744,745..3228,3229..3234,3235..3300,
                     3301..3303,3304..4818)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(715..741,742..744,745..3231,3232..3234,3235..4818)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             715..4818
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4831..4851
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6042..6062
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6072..6458
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2
                     /note="binds to a specific stem-loop structure in the viral
                     RNA (Peabody, 1993)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQNRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             6474..6623
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     ncRNA           7689..7725
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     misc_RNA        7746..7821
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        7826..7844
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     ncRNA           7851..7918
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     gene            8431..9550
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        8431..8774
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             8775..9347
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      9353..9550
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(10625..11213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10705..10724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttgatctt ttctacgggg tggagcgcaa aaaaccccgc ccctgacagg gcggggtttt
       61 ttcgcgatcg gagcggtact accattgaca ctacacatcc ttaattccaa tcctgtctgg
      121 cctccttcac cttttaacca tcttgcccat tccaactcgt gtcagattgc gtatcaagtg
      181 aaaaaaaaaa aattttaaat ctttaaccca atcaggtaat aactgtcgcc tcttttatct
      241 gccgcactgc atgaggtgtc cccttagtgg gaaagagtac tgagccaacc ctggaggaca
      301 gcaagggaaa aatacctaca acttgcttca taatggtcgt aaaaacaatc cttgtcggat
      361 ataagtgttg tagactgtcc cttatcctct gcgatgttct tcctctcaaa gtttgcgatt
      421 tctctctatc agaattgcca tcaagagact caggactaat ttcgcagtcc cacacgcact
      481 cgtacatgat tggctgaaat ttccctaaag aatttctttt tcacgaaaat ttttttttta
      541 cacaagattt tcagcagata taaaatggag agcaggacct ccgctgtgac tcttcttttt
      601 tttcttttat tctcactaca tacattttag ttattcgcca acatggacta caaagaccat
      661 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gctcatggac
      721 aagaagtact ccattgggct cgctatcggc acaaacagcg tcggctgggc cgtcattacg
      781 gacgagtaca aggtgccgag caaaaaattc aaagttctgg gcaataccga tcgccacagc
      841 ataaagaaga acctcattgg cgccctcctg ttcgactccg gggagacggc cgaagccacg
      901 cggctcaaaa gaacagcacg gcgcagatat acccgcagaa agaatcggat ctgctacctg
      961 caggagatct ttagtaatga gatggctaag gtggatgact ctttcttcca taggctggag
     1021 gagtcctttt tggtggagga ggataaaaag cacgagcgcc acccaatctt tggcaatatc
     1081 gtggacgagg tggcgtacca tgaaaagtac ccaaccatat atcatctgag gaagaagctt
     1141 gtagacagta ctgataaggc tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc
     1201 aaatttcggg gacacttcct catcgagggg gacctgaacc cagacaacag cgatgtcgac
     1261 aaactcttta tccaactggt tcagacttac aatcagcttt tcgaagagaa cccgatcaac
     1321 gcatccggag ttgacgccaa agcaatcctg agcgctaggc tgtccaaatc ccggcggctc
     1381 gaaaacctca tcgcacagct ccctggggag aagaagaacg gcctgtttgg taatcttatc
     1441 gccctgtcac tcgggctgac ccccaacttt aaatctaact tcgacctggc cgaagatgcc
     1501 aagcttcaac tgagcaaaga cacctacgat gatgatctcg acaatctgct ggcccagatc
     1561 ggcgaccagt acgcagacct ttttttggcg gcaaagaacc tgtcagacgc cattctgctg
     1621 agtgatattc tgcgagtgaa cacggagatc accaaagctc cgctgagcgc tagtatgatc
     1681 aagcgctatg atgagcacca ccaagacttg actttgctga aggcccttgt cagacagcaa
     1741 ctgcctgaga agtacaagga aattttcttc gatcagtcta aaaatggcta cgccggatac
     1801 attgacggcg gagcaagcca ggaggaattt tacaaattta ttaagcccat cttggaaaaa
     1861 atggacggca ccgaggagct gctggtaaag cttaacagag aagatctgtt gcgcaaacag
     1921 cgcactttcg acaatggaag catcccccac cagattcacc tgggcgaact gcacgctatc
     1981 ctcaggcggc aagaggattt ctaccccttt ttgaaagata acagggaaaa gattgagaaa
     2041 atcctcacat ttcggatacc ctactatgta ggccccctcg cccggggaaa ttccagattc
     2101 gcgtggatga ctcgcaaatc agaagagact atcactccct ggaacttcga ggaagtcgtg
     2161 gataaggggg cctctgccca gtccttcatc gaaaggatga ctaactttga taaaaatctg
     2221 cctaacgaaa aggtgcttcc taaacactct ctgctgtacg agtacttcac agtttataac
     2281 gagctcacca aggtcaaata cgtcacagaa gggatgagaa agccagcatt cctgtctgga
     2341 gagcagaaga aagctatcgt ggacctcctc ttcaagacga accggaaagt taccgtgaaa
     2401 cagctcaaag aggactattt caaaaagatt gaatgtttcg actctgttga aatcagcgga
     2461 gtggaggatc gcttcaacgc atccctggga acgtatcacg atctcctgaa aatcattaaa
     2521 gacaaggact tcctggacaa tgaggagaac gaggacattc ttgaggacat tgtcctcacc
     2581 cttacgttgt ttgaagatag ggagatgatt gaagaacgct tgaaaactta cgctcatctc
     2641 ttcgacgaca aagtcatgaa acagctcaag aggcgccgat atacaggatg ggggcggctg
     2701 tcaagaaaac tgatcaatgg gatccgagac aagcagagtg gaaagacaat cctggatttt
     2761 cttaagtccg atggatttgc caacaggaac ttcatgcagt tgatccatga tgactctctc
     2821 acctttaagg aggacatcca gaaagcacaa gtttctggcc agggggacag tctgcacgag
     2881 cacatcgcta atcttgcagg tagcccagct atcaaaaagg gaatactgca gaccgttaag
     2941 gtcgtggatg aactcgtcaa agtaatggga aggcataagc ccgagaatat cgttatcgag
     3001 atggcccgag agaaccaaac tacccagaag ggacagaaga acagtaggga aaggatgaag
     3061 aggattgaag agggtataaa agaactgggg tcccaaatcc ttaaggaaca cccagttgaa
     3121 aacacccagc ttcagaatga gaagctctac ctgtactacc tgcagaacgg cagggacatg
     3181 tacgtggatc aggaactgga catcaatcgg ctctccgact acgacgtgga tgccatcgtg
     3241 ccccagtctt ttctcaaaga tgattctatt gataataaag tgttgacaag atccgataaa
     3301 aatagaggga agagtgataa cgtcccctca gaagaagttg tcaagaaaat gaaaaattat
     3361 tggcggcagc tgctgaacgc caaactgatc acacaacgga agttcgataa tctgactaag
     3421 gctgaacgag gtggcctgtc tgagttggat aaagccggct tcatcaaaag gcagcttgtt
     3481 gagacacgcc agatcaccaa gcacgtggcc caaattctcg attcacgcat gaacaccaag
     3541 tacgatgaaa atgacaaact gattcgagag gtgaaagtta ttactctgaa gtctaagcta
     3601 gtctcagatt tcagaaagga ctttcagttt tataaggtga gagagatcaa caattaccac
     3661 catgcgcatg atgcctacct gaatgcagtg gtaggcactg cacttatcaa aaaatatccc
     3721 aagcttgaat ctgaatttgt ttacggagac tataaagtgt acgatgttag gaaaatgatc
     3781 gcaaagtctg agcaggaaat aggcaaggcc accgctaagt acttctttta cagcaatatt
     3841 atgaattttt tcaagaccga gattacactg gccaatggag agattcggaa gcgaccactt
     3901 atcgaaacaa acggagaaac aggagaaatc gtgtgggaca agggtaggga tttcgcgaca
     3961 gtccggaagg tcctgtccat gccgcaggtg aacatcgtta aaaagaccga agtacagacc
     4021 ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc
     4081 aaaaaagatt gggaccccaa gaaatacggc ggattcgatt ctcctacagt cgcttacagt
     4141 gtactggttg tggccaaagt ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa
     4201 ctgctgggca tcacaatcat ggagcgatca agcttcgaaa aaaaccccat cgactttctc
     4261 gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca ttaagcttcc caagtactct
     4321 ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa
     4381 ggtaacgagc tggcactgcc ctctaaatac gttaatttct tgtatctggc cagccactat
     4441 gaaaagctca aagggtcccc cgaagataat gagcagaagc agctgttcgt ggaacaacac
     4501 aaacactacc ttgatgagat catcgagcaa ataagcgaat tctccaaaag agtgatcctc
     4561 gccgacgcta acctcgataa ggtgctttct gcttacaata agcacaggga taagcccatc
     4621 agggagcagg cagaaaacat tatccacttg tttactctga ccaacttggg cgctcctgca
     4681 gccttcaagt acttcgacac caccatagac agaaagcggt acacctctac aaaggaggtc
     4741 ctggacgcca cactgattca tcagtcaatt acggggctct atgaaacaag aatcgacctc
     4801 tctcagctcg gtggagacag cagggctgac cccaagaaga agaggaaggt gtgagcttta
     4861 gtcaaatatt aatctatttc acctgttcaa actttactta atgtacaaat gtggtagtta
     4921 ttagttttgc aacggaactt gttccataat ctggtcctct gggacagcaa actgtctttc
     4981 actagtagcg ccagtttcgg gagtccacac agcattagtc accggtgcac cagcactaat
     5041 ctcacgacct tctgggtgtt taaatgggca gttagggttg cggcatccag ctgcaaactt
     5101 acaatcctca tcaattggat gagtgaaaaa acagtttggt ctggtacaac tgttgccttc
     5161 acgacacagt acaggagtag ttgcgtgacg tcttgggcac ttgtaattac ggcatgattt
     5221 accaaatcga cattgttcca aagccctctg ttgtttttgt tgtttctctt cttcggtgat
     5281 cttgtgttca ggtgatcgat gagcctttgg acagtccgga ttagagcgct ccggggagcg
     5341 cattatttgg gtcggctact gtcattggtt atgatggcct taccgagcac gaaggtggtt
     5401 ggatagacca caactggaaa cagttgtaca aacagattgc attcattttt gcttgtattg
     5461 gatactcgat ggccatcacc gctcttatct gtttcatcct caaccgtatt ccatttttgc
     5521 aactgcgagc ttcagaagag gctgaggaga aaggtatgga tgaggatcag attggagagt
     5581 tcgcttatga ctacgtggaa gtacgtcgtg attttttggc ttggggatca ggcccaaaca
     5641 atggcttcaa ggagccggaa gttctggatc aggtagttcc ggttaatgat ttcagcagtg
     5701 accagaatgt gactaatcag accaacgaat ctgagaagca gtagagtaaa tatagagatg
     5761 atatttagtg tattctaatg cttatgtaat gtattaagca aaaagttgtg tttatgagtt
     5821 agcatttgtc ttagcaaaca taaaattatg tcgacatttg caacccgcat gtctagtgtt
     5881 tttagatcga tcttcgatgt gtagaataat gcctccacgt gatgccccgc gattttgttg
     5941 ggtcgcaatg cctccaacat aaacccatca cgtataaaaa gccctcttaa ccctcccccc
     6001 tgtttcgttt gcttcatcac ttaacctgaa ctatcaacat gcccaaaaag aaaagaaaag
     6061 tgggtagtat ggcgtctaac ttcacccagt tcgttctggt tgacaacggt ggtaccggtg
     6121 acgttaccgt tgcgccgtct aacttcgcga acggtgttgc ggaatggatc tcttctaact
     6181 ctcgttctca ggcgtacaaa gttacctgct ctgttcgtca atcctccgcg cagaaccgta
     6241 aatacaccat caaagttgaa gttccgaaag ttgcgaccca gaccgttggt ggtgttgaac
     6301 tgccggttgc ggcgtggcgt tcttacctga acatggaact gaccatcccg atcttcgcga
     6361 ccaactctga ctgcgaactg atcgttaaag cgatgcaggg tctgctgaaa gacggtaacc
     6421 cgatcccgtc tgcgatcgcg gcgaactctg gtatctacgg ctccggacga gccgatgccc
     6481 ttgacgattt tgacttggac atgttgggtt ctgacgcttt ggatgacttt gatcttgata
     6541 tgcttggttc cgacgctcta gatgatttcg acttggatat gctgggatcc gatgccttgg
     6601 acgatttcga cttggatatg ttgatcaact gagcttctgt ttaattctgt aggataaacg
     6661 aaataactgt atctttataa tcatccttaa gatagccctg gactcgtcag acattagttc
     6721 gcgcacatta ttttcctcca ctgtctgcaa acagccacac ggcatttccc tctctagttt
     6781 catttatcaa agtagcacaa cacgctaatt ggaggacaat aagaagaaaa aaaaagaaaa
     6841 gcggtggggg agggattatt aaataaggat tatgtaaccc cagggtaccg ttctatacat
     6901 atttaaggat tatttaggac aatcgatgaa atcggcatca aactggatgg gagtatagtg
     6961 tccggataat cggataaatc atcttgcgag gagccgcttg gttggttggt gagaggagtg
     7021 aaatatgtgt ctcctcaccc aagaatcgcg atatcagcac cctgtggggg acactattgg
     7081 cctccctccc aaaccttcga tgtggtagtg ctttattata ttgattacat tgattacata
     7141 gctaaaccct gcctggttgc aagttgagct ccgaattcca atattagtaa aatgcctgca
     7201 agataacctc ggtatggcgt ccgaccccgc ttaattattt taactccttt ccaacgagga
     7261 cttcgtaatt tttgattagg gagttgagaa acggggggtc ttgatacctc ctcgatttca
     7321 gatcccaccc cctctcagtc ccaagtggga cccccctcgg ccgtgaaatg cgcgcacttt
     7381 agtttttttc gcatgtaaac gccggtgtcc gtcaattaaa agtcgcagac tagggtgaac
     7441 tttaccattt ttgtcgcact ccgtctcctc ggaatagggg tgtagtaatt ctgcagtagt
     7501 gcaattttta ccccgccaag ggggggcgaa aagagacgac ctcatcacgc attctccagt
     7561 cgctctctac gcctacagca ccgacgtagt taactttctc ccatatataa agcaattgcc
     7621 attcccctga aaactttaac ctctgctttt tcttgatttt tccttgccca aagaaaagca
     7681 tgaatgatct gatgagtccg tgaggacgaa acgagtaagc tcgtcatcat tgacagagtg
     7741 aatcagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     7801 aaagtggcac cgagtcggtg cgcgcacatg aggatcaccc atgtgctttt ggccggcatg
     7861 gtcccagcct cctcgctggc gccggctggg caacatgctt cggcatggcg aatgggacag
     7921 ctttggactg cttttagtgt acatctgata atatagtact accacgtatg ataatgtaga
     7981 gaataggctt ccttgtcgag tgtgtttgca gttttcttga gtttcaaggt ttaaatgctg
     8041 gtatattagt tcatcgaagg tttcagccaa tagcacctta aatcaatcaa actaattcga
     8101 ctcttacgaa agagcctact gtgtttagta tcgaagtcgt ttacctttca tgttgaatag
     8161 cttcctctct gaccctaaca tttcaagatc ctcctaaagt tacccggatt gtgaaattct
     8221 aatgatccac ctgcccaatg cattttttct ttattcagtt tacctttttt acctaatata
     8281 cgagcttgtt aaagtaagtg gcactgcaat actaggctta ttgttgatat tatgatgaat
     8341 cgttttcaca aacttgattt cctgtgaact caccatgtac taaggaaaaa aacatgcatc
     8401 accatctgaa tatttgaccg ctagctcatg gacatggagg cccagaatac cctccttgac
     8461 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     8521 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     8581 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     8641 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     8701 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     8761 gaacataaac aaccatgggt accactcttg acgacacggc ttaccggtac cgcaccagtg
     8821 tcccggggga cgccgaggcc atcgaggcac tggatgggtc cttcaccacc gacaccgttt
     8881 ttcgcgtcac cgccaccggg gacggcttca ccctgcggga ggtgccggtg gacccgcccc
     8941 tgaccaaggt gttccccgac gacgaatcgg acgacgaatc ggacgacggg gaggacggcg
     9001 acccggactc ccggacgttc gtcgcgtacg gggacgacgg cgacctggcg ggcttcgtgg
     9061 tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga ggacatcgag gtcgccccgg
     9121 agcaccgggg gcacggggtc gggcgcgcgt tgatggggct cgcgacggag ttcgcccgcg
     9181 agcggggcgc cgggcacctc tggctggagg tcaccaacgt caacgcaccg gcgatccacg
     9241 cgtaccggcg gatggggttc accctctgcg gcctggacac cgccctgtac gacggcaccg
     9301 cctcggacgg cgagcaggcg ctctacatga gcatgccctg cccctaatca gtactgacaa
     9361 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
     9421 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
     9481 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
     9541 cgctatactg gcttcatacc cagcattgac ctttggtatg agcatctgaa aaacaaccag
     9601 gtgttgcaaa gttaaacatc cttctttgtt catatagaac ccactattca tggtactccc
     9661 caatcgaatt tcacattctg gttttgaaat tacacaccac gttagcttat aagatttcat
     9721 ataacttatt gatatacggt ttccattgtt cgaatagttg aggttgtatg taattcgatt
     9781 gaaggggcca tttttgtttc ctacttttcc tgggagctta tccgatgcgc ttcaaagctg
     9841 gaattgtaaa tatagagaaa aagaaggatg ttgttttatt cttgaaagag tataatttta
     9901 cttctagcaa ctctcccact tcgcttgact tcatttattt cttgggcaca taggcgtagt
     9961 aatctagacc aacagataat ttgccggaat gatatagcga ttggaaaatg aactgaaatt
    10021 ttttgctgtc tttcaatttg acgggcagtt catcaggcgc gccgtgaccg accatataaa
    10081 tacgttgaga atgttattct tcctcgtagt tgaagtggct tcataatttc agaactcaat
    10141 agataaacta ggatgtttta aagcaattaa tgctcacaag taaggagcga ctctcttgct
    10201 tttcgaatac taaaagtatc gtcccaaccc agaaaaaaag acctcttaac tgcaaaataa
    10261 actctatata tttcttctaa aacagtttca ggttggatag tatcgcattc tcatcacttc
    10321 taactagtag gccatgagat atattaacgt ttacttgagt tctaagttct ccgaattaga
    10381 tgcacagcac aaacaagatt aggtttcact tggtacaaaa tacgaacaga gtttaaggtc
    10441 gtaatttcat ttcgttattg atccccacaa tctattctta tcacagtcat cagatagtcg
    10501 cgaaaaagca tgcagaaaag ggggtcgtcc ctatctaagt tgtagcatta caacaaatat
    10561 gaccgctgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
    10621 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
    10681 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
    10741 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
    10801 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
    10861 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
    10921 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    10981 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    11041 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    11101 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    11161 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
//
