LOCUS       sequence-269821-       12654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrates CRISPRi effector and tet-responsive regulator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12654)
  AUTHORS   McGlincy NJ, Meacham ZA, Reynaud KK, Muller R, Baum R, Ingolia NT
  TITLE     A genome-scale CRISPR interference guide library enables 
            comprehensive phenotypic profiling in yeast.
  JOURNAL   BMC Genomics. 2021 Mar 23;22(1):205. doi: 
            10.1186/s12864-021-07518-0.
  PUBMED    33757429
REFERENCE   2  (bases 1 to 12654)
  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: "BMC 
            Genomics."; date: "2021-03-23"; volume: "22(1):205. doi"; pages: " 
            10.1186/s12864-021-07518-0"
COMMENT     Sequence Label: pNTI647 dCas9-Mxi1 TetR KanMX
FEATURES             Location/Qualifiers
     source          1..12654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(97..888,889..957))
                     /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     720..739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(958..1049)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1116..1135
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1342..1375
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     terminator      complement(2092..2339)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2305..2323
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2363..2485)
                     /codon_start=1
                     /product="transcriptional repressor domain from the mouse 
                     Max-interacting protein 1 (Schreiber-Agus et al., 1995)"
                     /label=Mxi1
                     /translation="MERVRMINVQRLLEAAEFLERRERECEHGYASSFPSMPSPR"
     CDS             complement(2519..2539)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(2561..4075,4076..4078,4079..4144,4145..4150,
                     4151..6634,6635..6637,6638..6664))
                     /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             complement(join(2561..4144,4145..4147,4148..6634,6635..6637,
                     6638..6664))
                     /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             complement(2561..6664)
                     /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             complement(6674..6694)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(6717..7115)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             7928..8551
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(8001..8020)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    8960..8993
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            9048..10404
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        9048..9391
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             9392..10201
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      10207..10404
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    10467..10500
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(10543..10562)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10544..10562)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10572..10588)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10581..10603)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(11250..11283)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(11753..11769)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11868..11885)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12039..12627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12119..12138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctgacgctc agtggaacga cgcgcgcgta actcacgtta agggattttg gtcatgagct
       61 tgcgccgtcc cgtcaagtca gcgtaatgct ctgcttttac caatgcttaa tcagtgaggc
      121 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
      181 gataactacg atacgggagg gcttaccatc tggccccagc gctgcgatga taccgcgaga
      241 accacgctca ccggctccgg atttatcagc aataaaccag ccagccggaa gggccgagcg
      301 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
      361 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg ctacaggcat
      421 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
      481 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
      541 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
      601 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
      661 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
      721 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
      781 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
      841 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
      901 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatatt
      961 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1021 atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca attaaccaat
     1081 tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc ccttgtttgc
     1141 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     1201 gtagcgccga tggtagtgtg gggactcccc atgcgagagt agggaactgc caggcatcaa
     1261 ataaaacgaa aggctcagtc gaaagactgg gcctttcgcc cgggctaatt atggggtgtc
     1321 gcccttattc gactctatag tgaagttcct attctctaga aagtatagga acttctgaag
     1381 tggggattta aatgcggccg cgctgagggt ttaatcggca tgcaaacatc tacacaatta
     1441 gcaagggcaa tccatatttt gtcttttcgc gccctggaaa ggcctaagta atgtcgtaaa
     1501 cgcattctat ctgtacttca actctcctct gtgcattggt ttgtgcaaat cacattttac
     1561 gatactgcca gatatatgca aaaagagaaa accaagggac cagaacaaag caaaattacg
     1621 atattcttcg aattccttcg tgcttgacta agacaaaggg atggacgtag cgatttttag
     1681 cgggccaaga actggttccg aaaaagcaca ggtacaccga accctcagct aaggagggac
     1741 agcaccgatg cggaaggaca aactttcttt ttgcctatca cagtatctta tcgagctaac
     1801 tattttcgac acacatgaaa aagcagaaat attaacgaaa aagaaaagaa agaccatgtc
     1861 atgtacgggc aatcagaatc tgtaacaagc gccatttttt tttctgtatc gggccctcct
     1921 tactgctctc cttccgtgta acgcgttatg attaagtcct cagcgagctc gcatggaatg
     1981 cgtgcgatga gcgacctcat gctatacctg agaaagcaac ctgacctaca ggaaagagtt
     2041 actcaagaat aagaattttc gttttaaaac ctaagagtca ctttaaaatt tgcaaattaa
     2101 agccttcgag cgtcccaaaa ccttctcaag caaggttttc agtataatgt tacatgcgta
     2161 cacgcgtctg tacagaaaaa aaagaaaaat ttgaaatata aataacgttc ttaatactaa
     2221 cataactata aaaaaataaa tagggaccta gacttcaggt tgtctaactc cttccttttc
     2281 ggttagagcg gatgtggggg gagggcgtga atgtaagcgt gacataacta attacatgac
     2341 tcgagaagag ctcgagttag cctctgggag agggcatgct agggaaagaa ctggcatacc
     2401 cgtgttcgca ttctctttct cttctttcta aaaactctgc ggcttctaac agcctttgca
     2461 cattaatcat tctcacacgt tccatgctgc cgccgctgcc gccaagcttg gaactaccta
     2521 ccttgcgctt tttcttggga gctccctcgt cagccctgct gtctccaccg agctgagaga
     2581 ggtcgattct tgtttcatag agccccgtaa ttgactgatg aatcagtgtg gcgtccagga
     2641 cctcctttgt agaggtgtac cgctttctgt ctatggtggt gtcgaagtac ttgaaggctg
     2701 caggcgcgcc caagttggtc agagtaaaca agtggataat gttttctgcc tgctccctga
     2761 tgggcttatc cctgtgctta ttgtaagcag aaagcacctt atcgaggtta gcgtcggcga
     2821 ggatcactct tttggagaat tcgcttattt gctcgatgat ctcatcaagg tagtgtttgt
     2881 gttgttccac gaacagctgc ttctgctcat tatcttcggg agaccctttg agcttttcat
     2941 agtggctggc cagatacaag aaattaacgt atttagaggg cagtgccagc tcgttacctt
     3001 tctgcagctc gcccgcacta gcgagcattc gtttccggcc gttttcaagc tcaaagagag
     3061 agtacttggg aagcttaatg atgaggtctt ttttgacctc tttatatcct ttcgcctcga
     3121 gaaagtcgat ggggtttttt tcgaagcttg atcgctccat gattgtgatg cccagcagtt
     3181 ccttgacgct tttgagtttt ttagacttcc ctttctccac tttggccaca accagtacac
     3241 tgtaagcgac tgtaggagaa tcgaatccgc cgtatttctt ggggtcccaa tcttttttgc
     3301 gtgcgatcag cttgtcgctg ttccttttcg ggaggatact ttccttggag aagcctccgg
     3361 tctgtacttc ggtcttttta acgatgttca cctgcggcat ggacaggacc ttccggactg
     3421 tcgcgaaatc cctacccttg tcccacacga tttctcctgt ttctccgttt gtttcgataa
     3481 gtggtcgctt ccgaatctct ccattggcca gtgtaatctc ggtcttgaaa aaattcataa
     3541 tattgctgta aaagaagtac ttagcggtgg ccttgcctat ttcctgctca gactttgcga
     3601 tcattttcct aacatcgtac actttatagt ctccgtaaac aaattcagat tcaagcttgg
     3661 gatatttttt gataagtgca gtgcctacca ctgcattcag gtaggcatca tgcgcatggt
     3721 ggtaattgtt gatctctctc accttataaa actgaaagtc ctttctgaaa tctgagacca
     3781 gcttagactt cagagtaata actttcacct ctcgaatcag tttgtcattt tcatcgtact
     3841 tggtgttcat gcgtgaatcg agaatttggg ccacgtgctt ggtgatctgg cgtgtctcaa
     3901 caagctgcct tttgatgaag ccggctttat ccaactcaga caggccacct cgttcagcct
     3961 tagtcagatt atcgaacttc cgttgtgtga tcagtttggc gttcagcagc tgccgccaat
     4021 aatttttcat tttcttgaca acttcttctg aggggacgtt atcactcttc cctctatttt
     4081 tatcggatct tgtcaacact ttattatcaa tagaatcatc tttgagaaaa gactggggca
     4141 cgatagcatc cacgtcgtag tcggagagcc gattgatgtc cagttcctga tccacgtaca
     4201 tgtccctgcc gttctgcagg tagtacaggt agagcttctc attctgaagc tgggtgtttt
     4261 caactgggtg ttccttaagg atttgggacc ccagttcttt tataccctct tcaatcctct
     4321 tcatcctttc cctactgttc ttctgtccct tctgggtagt ttggttctct cgggccatct
     4381 cgataacgat attctcgggc ttatgccttc ccattacttt gacgagttca tccacgacct
     4441 taacggtctg cagtattccc tttttgatag ctgggctacc tgcaagatta gcgatgtgct
     4501 cgtgaagact gtccccctgg ccagaaactt gtgctttctg gatgtcctcc ttaaaggtga
     4561 gagagtcatc atggatcaac tgcatgaagt tccggttggc aaatccatcg gacttaagaa
     4621 aatccaggat tgtctttcca ctctgcttgt ctcggatccc attgatcagt tttcttgaca
     4681 gccgccccca tcctgtatat cggcgcctct tgagctgttt catgactttg tcgtcgaaga
     4741 gatgagcgta agttttcaag cgttcttcaa tcatctccct atcttcaaac aacgtaaggg
     4801 tgaggacaat gtcctcaaga atgtcctcgt tctcctcatt gtccaggaag tccttgtctt
     4861 taatgatttt caggagatcg tgatacgttc ccagggatgc gttgaagcga tcctccactc
     4921 cgctgatttc aacagagtcg aaacattcaa tctttttgaa atagtcttct ttgagctgtt
     4981 tcacggtaac tttccggttc gtcttgaaga ggaggtccac gatagctttc ttctgctctc
     5041 cagacaggaa tgctggcttt ctcatccctt ctgtgacgta tttgaccttg gtgagctcgt
     5101 tataaactgt gaagtactcg tacagcagag agtgtttagg aagcaccttt tcgttaggca
     5161 gatttttatc aaagttagtc atcctttcga tgaaggactg ggcagaggcc cccttatcca
     5221 cgacttcctc gaagttccag ggagtgatgg tctcttctga tttgcgagtc atccacgcga
     5281 atctggaatt tccccgggcg agggggccta catagtaggg tatccgaaat gtgaggattt
     5341 tctcaatctt ttccctgtta tctttcaaaa aggggtagaa atcctcttgc cgcctgagga
     5401 tagcgtgcag ttcgcccagg tgaatctggt gggggatgct tccattgtcg aaagtgcgct
     5461 gtttgcgcaa cagatcttct ctgttaagct ttaccagcag ctcctcggtg ccgtccattt
     5521 tttccaagat gggcttaata aatttgtaaa attcctcctg gcttgctccg ccgtcaatgt
     5581 atccggcgta gccattttta gactgatcga agaaaatttc cttgtacttc tcaggcagtt
     5641 gctgtctgac aagggccttc agcaaagtca agtcttggtg gtgctcatca tagcgcttga
     5701 tcatactagc gctcagcgga gctttggtga tctccgtgtt cactcgcaga atatcactca
     5761 gcagaatggc gtctgacagg ttctttgccg ccaaaaaaag gtctgcgtac tggtcgccga
     5821 tctgggccag cagattgtcg agatcatcat cgtaggtgtc tttgctcagt tgaagcttgg
     5881 catcttcggc caggtcgaag ttagatttaa agttgggggt cagcccgagt gacagggcga
     5941 taagattacc aaacaggccg ttcttcttct ccccagggag ctgtgcgatg aggttttcga
     6001 gccgccggga tttggacagc ctagcgctca ggattgcttt ggcgtcaact ccggatgcgt
     6061 tgatcgggtt ctcttcgaaa agctgattgt aagtctgaac cagttggata aagagtttgt
     6121 cgacatcgct gttgtctggg ttcaggtccc cctcgatgag gaagtgtccc cgaaatttga
     6181 tcatatgcgc cagcgcgaga tagatcaacc gcaagtcagc cttatcagta ctgtctacaa
     6241 gcttcttcct cagatgatat atggttgggt acttttcatg gtacgccacc tcgtccacga
     6301 tattgccaaa gattgggtgg cgctcgtgct ttttatcctc ctccaccaaa aaggactcct
     6361 ccagcctatg gaagaaagag tcatccacct tagccatctc attactaaag atctcctgca
     6421 ggtagcagat ccgattcttt ctgcgggtat atctgcgccg tgctgttctt ttgagccgcg
     6481 tggcttcggc cgtctccccg gagtcgaaca ggagggcgcc aatgaggttc ttctttatgc
     6541 tgtggcgatc ggtattgccc agaactttga attttttgct cggcaccttg tactcgtccg
     6601 taatgacggc ccagccgacg ctgtttgtgc cgatagcgag cccaatggag tacttcttgt
     6661 ccatcccggg tctaactttt ctcttcttct ttggggctct agacatctct agaaaactta
     6721 gattagattg ctatgctttc tttctaatga gcaagaagta aaaaaagttg taatagaaca
     6781 agaaaaatga aactgaaact tgagaaattg aagaccgttt attaacttaa atatcaatgg
     6841 gaggtcatcg aaagagaaaa aaatcaaaaa aaaaattttc aagaaaaaga aacgtgataa
     6901 aaatttttat tgcctttttc gacgaagaaa aagaaacgag gcggtctctt ttttcttttc
     6961 caaaccttta gtacgggtaa ttaacgacac cctagaggaa gaaagagggg aaatttagta
     7021 tgctgtgctt gggtgttttg aagtggtacg gcgatgcgcg gagtccgaga aaatctggaa
     7081 gagtaaaaaa ggagtagaaa cattttgaag ctatgcgctt atttagaagt gtcaacaacg
     7141 tatctaccaa cggaatgcgt gcgatcgcgt gcattctagt cgtgcaatgt atgactttaa
     7201 gatttgtgag caggaagaaa agggagaatc ttctaacgat aaacccttga aaaactgggt
     7261 agactacgct atgttgagtt gctacgcagg ctgcacaatt acacgagaat gctcccgcct
     7321 aggatttaag gctaagggac gtgcaatgca gacgacagat ctaaatgacc gtgtcggtga
     7381 agtgttcgcc aaacttttcg gttaacacat gcagtgatgc acgcgcgatg gtgctaagtt
     7441 acatatatat atatatgtat agccatagtg atgtctaagt aacctttatg gtatatttct
     7501 taatgtggaa agatactagc gcgcgcaccc acacacaagc ttcgtctttt cttgaagaaa
     7561 agaggaagct cgctaaatgg gattccactt tccgttccct gccagctgat ggaaaaaggt
     7621 tagtggaacg atgaagaata aaaagagaga tccactgagg tgaaatttca gctgacagcg
     7681 agtttcatga tcgtgatgaa caatggtaac gagttgtggc tgttgccagg gagggtggtt
     7741 ctcaactttt aatgtatggc caaatcgcta cttgggtttg ttatataaca aagaagaaat
     7801 aatgaactga ttctcttcct ccttcttgtc ctttcttaat tctgttgtaa ttaccttcct
     7861 ttgtaatttt ttttgtaatt attcttctta ataatccaaa caaacacaca tattacaata
     7921 tattaaaatg tctagattag ataaaagtaa agtgattaac agcgcattag agctgcttaa
     7981 tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca
     8041 gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga
     8101 gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt
     8161 tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa
     8221 agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc
     8281 ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca gcgctgtggg
     8341 gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag
     8401 ggaaacacct actactgata gtatgccgcc attattacga caagctatcg aattatttga
     8461 tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga
     8521 aaaacaactt aaatgtgaaa gtgggtctta agtctgaaga atgaatgatt tgatgatttc
     8581 tttttccctc catttttctt actgaatata tcaatgatat agacttgtat agtttattat
     8641 ttcaaattaa gtagctatat atagtcaaga taacgtttgt ttgacacgat tacattattc
     8701 gtcgacatct tttttcagcc tgtcgtggta gcaatttgag gagtattatt aattgaatag
     8761 gttcattttg cgctcgcata aacagttttc gtcagggaca gtatgttgga atgagtggta
     8821 attaatggtg acatgacatg ttatagcaat aaccttgatg tttacatcgt agtttaatgt
     8881 acaccccgcg aattcgttca agtaggagtg caccaattgc aaagggaaaa gctgaatggg
     8941 cagttcgaaa acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtct
     9001 agagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc
     9061 agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc
     9121 cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc
     9181 gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc
     9241 ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt
     9301 aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca
     9361 gttctcacat cacatccgaa cataaacaac catgggtaag gaaaagactc acgtttcgag
     9421 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     9481 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
     9541 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     9601 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     9661 tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca gcattccagg tattagaaga
     9721 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttaca
     9781 ttcgattcct gtttgtaatt gtccttttaa cagcgaccgt gtatttcgtc tcgctcaggc
     9841 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     9901 ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc
     9961 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
    10021 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
    10081 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
    10141 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
    10201 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
    10261 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
    10321 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
    10381 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtgtcg
    10441 aaaacgagct ctcgagaacc cttaatataa cttcgtataa tgtatgctat acgaagttat
    10501 taggtgatat cagatccact agtggcctat gcacccaatt cgccctatag tgagtcgtat
    10561 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
    10621 cctgcaggac tagtgctgag gcattaatct actatcggcg actctctcga aatttttctt
    10681 aacgcgtcct tgtactgcgt ctaacgcttt tgccacttgg atttctatta taggaaatag
    10741 tctcacttac tgggcgacga attttcgcgt tttgatgaag cacaggaaga atttcttttt
    10801 tttttggctt cttctggttc cgttttttac gcgcacaaat ctaaaaaaag aaataattat
    10861 aacctagtct cgaaaatttt catcgatcca ttcgttcctt tttttcgatt ttttcagatc
    10921 aaaattcttg tttctttctt tgtcttagtt tatattaaaa gatattttga ttttactcct
    10981 gaactattta ttctttctaa gaaggccaga acactacagc tgttttaacc gactacgaag
    11041 ttctccattc tcgaacacta gccttcattt accaaacagg aactagcgta tatcattagt
    11101 ccttattcga aaagagattg gtagatattt attgtagttt gtgagaagga gaaaatactg
    11161 tcattggact gatagttaga ggacattaac ctctcttacg ttcgctcatt aagacctcag
    11221 cgcggccgca aatttaaata aaatgaagtg aagttcctat actttctaga gaataggaac
    11281 ttctatagtg agtcgaataa gggcgacaca aaatttattc taaatgcata ataaatactg
    11341 ataacatctt atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat
    11401 caaaaactga ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa
    11461 ctagaaatat tgtatataca aaaaatcata aataatagat gaatagttta attataggtg
    11521 ttcatcaatc gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata
    11581 aggttaaata attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa
    11641 tgctctttcc ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt
    11701 gcggattaac gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg
    11761 tcgttttaca acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt
    11821 ctgcttagtt tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac
    11881 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
    11941 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
    12001 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
    12061 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
    12121 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
    12181 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
    12241 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
    12301 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    12361 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    12421 tatgtaggcg gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga
    12481 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
    12541 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
    12601 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac gggg
//
