LOCUS       sequence-222978-       11156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 driven by PGK1 promoter. gRNA can be cloned into the BplI 
            sites. URA3MX marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11156)
  AUTHORS   
  TITLE     Cas9-mediated gene editing in Saccharomyces cerevisiae
  JOURNAL   Protocol Exchange (2017)
REFERENCE   2  (bases 1 to 11156)
  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: "Protocol 
            Exchange (2017)"
COMMENT     Sequence Label: pJH2972
FEATURES             Location/Qualifiers
     source          1..11156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(284..481)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(487..1290)
                     /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(1291..1634)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2652..6755
                     /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             2652..6755
                     /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             6768..6788
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6792..6812
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6816..6836
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6837..6902
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      6921..7168
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6937..6955)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(7228..7683)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7365..7386)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7577..7596
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7809..7831
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7823..7840
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7824..7840
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7847..7866
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7847..7865
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7898..7914
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7898..7914
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        7946..8214
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        8242..8316
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      8321..8340
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        complement(8383..8401)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8422..8438)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8422..8438)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8435..8457)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8446..8462
                     /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)."
     promoter        complement(join(8470..8476,8477..8494,8495..8500))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8515..8536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8653..8670)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8824..9412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8904..8923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9583..10374,10375..10443))
                     /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     10206..10225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10444..10548)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    10585..11088
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     11129..11147
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcgaattcg
      241 agctcgttta aactggatgg cggcgttagt atcgaatcga cagcagtata gcgaccagca
      301 ttcacatacg attgacgcat gatattactt tctgcgcact taacttcgca tctgggcaga
      361 tgatgtcgag gcgaaaaaaa atataaatca cgctaacatt tgattaaaat agaacaacta
      421 caatataaaa aaactataca aatgacaagt tcttgaaaac aagaatcttt ttattgtcag
      481 tactgattag ttttgctggc cgcatcttct caaatatgct tcccagcctg cttttctgta
      541 acgttcaccc tctaccttag catcccttcc ctttgcaaat agtcctcttc caacaataat
      601 aatgtcagat cctgtagaga ccacatcatc cacggttcta tactgttgac ccaatgcgtc
      661 tcccttgtca tctaaaccca caccgggtgt cataatcaac caatcgtaac cttcatctct
      721 tccacccatg tctctttgag caataaagcc gataacaaaa tctttgtcgc tcttcgcaat
      781 gtcaacagta cccttagtat attctccagt agatagggag cccttgcatg acaattctgc
      841 taacatcaaa aggcctctag gttcctttgt tacttcttct gccgcctgct tcaaaccgct
      901 aacaatacct gggcccacca caccgtgtgc attcgtaatg tctgcccatt ctgctattct
      961 gtatacaccc gcagagtact gcaatttgac tgtattacca atgtcagcaa attttctgtc
     1021 ttcgaagagt aaaaaattgt acttggcgga taatgccttt agcggcttaa ctgtgccctc
     1081 catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt tgggacctaa
     1141 tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt
     1201 tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc
     1261 agcacgttcc ttatatgtag ctttcgacat ggttgtttat gttcggatgt gatgtgagaa
     1321 ctgtatccta gcaagatttt aaaaggaagt atatgaaaga agaacctcag tggcaaatcc
     1381 taacctttta tatttctcta caggggcgcg gcgtggggac aattcaacgc gtctgtgagg
     1441 ggagcgtttc cctgctcgca ggtccgcagc gaggagccgt aatttttgct tcgcgccgtg
     1501 cggccatcaa aatgtatgga tgcaaatgat tatacatggg gatgtatggg ctaaatgtac
     1561 gggcgacagt cacatcatgc ccctgagctg cgcacgtcaa gactgtcaag gagggtattc
     1621 tgggcctcca tgtcgctggc cgggtgaccc ggcggggaaa gtaccttcaa agaatggggt
     1681 cttatcttgt tttgcaagta ccactgagca ggataataat agaaatgata atatactata
     1741 gtagagataa cgtcgatgac ttcccatact gtaattgctt ttagttgtgt atttttagtg
     1801 tgcaagtttc tgtaaatcga ttaatttttt tttctttcct ctttttatta accttaattt
     1861 ttattttaga ttcctgactt caactcaaga cgcacagata ttataacatc tgcataatag
     1921 gcatttgcaa gaattactcg tgagtaagga aagagtgagg aactatcgca tacctgcatt
     1981 taaagatgcc gatttgggcg cgaatccttt attttggctt caccctcata ctattatcag
     2041 ggccagaaaa aggaagtgtt tccctccttc ttgaattgat gttaccctca taaagcacgt
     2101 ggcctcttat cgagaaagaa attaccgtcg ctcgtgattt gtttgcaaaa agaacaaaac
     2161 tgaaaaaacc cagacacgct cgacttcctg tcttcctatt gattgcagct tccaatttcg
     2221 tcacacaaca aggtcctagc gacggctcac aggttttgta acaagcaatc gaaggttctg
     2281 gaatggcggg aaagggttta gtaccacatg ctatgatgcc cactgtgatc tccagagcaa
     2341 agttcgttcg atcgtactgt tactctctct ctttcaaaca gaattgtccg aatcgtgtga
     2401 caacaacagc ctgttctcac acactctttt cttctaacca agggggtggt ttagtttagt
     2461 agaacctcgt gaaacttaca tttacatata tataaacttg cataaattgg tcaatgcaag
     2521 aaatacatat ttggtctttt ctaattcgta gtttttcaag ttcttagatg ctttcttttt
     2581 ctctttttta cagatcatca aggaagtaat tatctacttt ttacaacaaa tataaaacac
     2641 cggatctcaa aatggataaa aagtatagta ttggtttaga tattggtact aactctgtgg
     2701 gttgggcagt tatcactgac gaatataaag ttccatcaaa gaaatttaag gtgttaggta
     2761 acactgacag acactcaata aaaaagaatc ttatcggtgc tcttttgttc gactccggtg
     2821 aaactgccga ggctacacgt ttaaaaagaa cagcaagaag aagatatacc cgtagaaaaa
     2881 atagaatatg ttatttacaa gaaatctttt ctaatgaaat ggctaaagtt gatgattcct
     2941 ttttccatag attggaagag tcatttttgg ttgaagaaga caaaaagcat gagagacatc
     3001 caatctttgg gaatatagtt gatgaagtgg cttaccatga aaaatatcct accatttatc
     3061 atttaagaaa gaaattggta gattcaactg ataaagctga ccttagatta atctatttag
     3121 cacttgccca tatgattaaa tttagaggtc attttttgat tgaaggtgat ttgaacccag
     3181 ataattctga cgtggataaa ttatttattc aattagtcca aacctacaac caattatttg
     3241 aggaaaatcc aattaatgct agtggtgtcg atgccaaagc tatattatca gccagattat
     3301 caaaatctag acgtttggaa aatttgattg cccaattgcc aggagaaaaa aagaatggat
     3361 tatttggaaa cttgatcgca ttatcattgg gtttgacacc aaattttaaa tctaattttg
     3421 atttagctga agatgctaaa ttacaattat caaaagacac ctatgacgac gatttggaca
     3481 atttacttgc tcaaattggt gatcaatatg cagatttgtt cttagctgct aaaaacttat
     3541 ctgatgctat tttgttgtct gatattttga gagtgaacac agaaataacc aaagctccat
     3601 tatcagcatc tatgatcaaa cgttatgatg aacaccatca ggatttgact ttattgaaag
     3661 ctttggtgag acaacaattg ccagagaagt ataaagaaat ctttttcgat caatctaaaa
     3721 acgggtatgc aggttatatt gatgggggtg cctcccaaga ggaattttac aaatttataa
     3781 aacctatttt agaaaagatg gatgggactg aggaactttt ggtcaaattg aacagagaag
     3841 atttgttacg taaacagaga acttttgata atggtagtat acctcaccaa attcatttgg
     3901 gtgagttgca tgcaatttta agaagacaag aagattttta tccattttta aaagataata
     3961 gagaaaaaat cgagaaaatt ttaaccttta gaattccata ctatgttggg cctttggcta
     4021 gaggtaattc aagatttgcc tggatgacac gtaaatcaga agaaactatt accccttgga
     4081 attttgaaga ggttgttgat aaaggagcat cagcacagag ttttattgaa agaatgacca
     4141 atttcgataa aaacttacca aatgaaaaag ttttaccaaa acattccttg ttatacgaat
     4201 attttactgt ttacaatgaa cttacaaagg ttaaatatgt tactgaaggt atgcgtaagc
     4261 cagccttttt atctggagaa cagaaaaagg caatagttga tttattgttt aaaacaaata
     4321 gaaaagttac tgttaaacaa ttaaaagaag attactttaa gaaaattgaa tgttttgatt
     4381 cagttgaaat cagtggtgtt gaagacagat ttaatgctag tttaggaact taccatgatt
     4441 tacttaaaat tatcaaagat aaagatttct tggataacga agaaaatgaa gacattttag
     4501 aagacattgt tttaacctta actttattcg aagatagaga gatgattgaa gaacgtttga
     4561 agacttatgc acatttgttt gacgataaag tgatgaaaca gttgaaaaga agacgttata
     4621 ctggatgggg tagattgtct cgtaaattga tcaatggaat tagagataaa caaagtggta
     4681 aaactatctt ggactttttg aaatctgacg gatttgctaa tagaaatttc atgcaattga
     4741 tccacgacga tagtttgaca tttaaagaag acatccaaaa ggcccaagtg agtgggcaag
     4801 gtgattcatt acatgaacat attgcaaatt tagccggatc tcctgctatt aagaaaggga
     4861 tattacaaac tgttaaagtt gtggatgaat tagtgaaagt aatgggaaga cataaacctg
     4921 aaaacattgt cattgagatg gcaagagaaa atcaaactac acaaaaagga cagaaaaata
     4981 gtagagaacg tatgaaaaga atagaagagg gtattaaaga attgggtagt caaatattga
     5041 aagaacaccc agtggaaaat acccagttgc aaaatgaaaa attatatctt tactaccttc
     5101 aaaatggacg tgatatgtat gttgatcagg aattagatat aaatagactt tcagattatg
     5161 atgtagatca tatagttcca caatctttct tgaaagatga ttccatagac aataaagtat
     5221 taactagaag tgataaaaat agaggtaaaa gtgataatgt cccaagtgag gaagtcgtca
     5281 aaaagatgaa aaattactgg cgtcaacttt tgaatgctaa attaattact caaagaaaat
     5341 ttgataattt gactaaagca gaaagaggtg ggctttctga attagataaa gccgggttca
     5401 ttaaaagaca attggtcgaa actagacaaa ttactaaaca tgttgcccaa attttagatt
     5461 cccgtatgaa cactaagtat gacgaaaatg ataagttaat acgtgaggtt aaagtcatta
     5521 ctttaaaatc aaaacttgtc tctgatttca gaaaggattt ccaattctat aaagttagag
     5581 aaattaataa ttatcatcat gctcatgatg catatttgaa tgctgtagtt ggaactgctt
     5641 taatcaagaa ataccctaaa ttagaatctg aatttgtata tggtgattac aaagtctatg
     5701 atgttagaaa gatgattgct aaatcagaac aagaaattgg taaagctaca gctaaatact
     5761 tcttttactc taacattatg aatttcttta aaacagaaat tactttggca aacggtgaaa
     5821 ttagaaaaag acctcttatt gaaacaaatg gtgagactgg agagatagtt tgggacaaag
     5881 ggcgtgattt cgctactgtt agaaaagttt tatcaatgcc acaagttaac attgtaaaga
     5941 aaacagaggt tcaaactggt ggtttctcaa aagaaagtat tttgcctaaa agaaatagtg
     6001 ataaattgat tgccagaaaa aaggattggg atccaaagaa atatggtggt ttcgactcac
     6061 caaccgtagc ctattctgtt ttggttgtgg caaaggttga aaagggtaaa agtaaaaagc
     6121 ttaaatcagt aaaagaactt ttgggtatta caataatgga aagaagttcc tttgaaaaga
     6181 accctattga ttttttggaa gctaaaggtt ataaggaagt aaagaaggac ttaataatca
     6241 aattgcctaa atattcttta tttgaattag aaaatgggag aaaaagaatg ttggcttctg
     6301 ctggagaatt gcaaaagggt aatgaattag cattgccttc caaatatgtt aacttcttgt
     6361 atttagcttc acactatgaa aagttgaaag ggtcaccaga agataacgag caaaaacaat
     6421 tatttgttga acaacacaaa cactacttag atgagattat agaacaaatt agtgaattca
     6481 gtaaaagagt gatattagct gatgcaaatt tagataaagt tttgtcagcc tataacaaac
     6541 atagagataa gccaattaga gaacaagcag aaaacattat tcacttattt acccttacca
     6601 atttaggagc acctgctgct ttcaagtatt ttgatacaac aattgatcgt aaaagatata
     6661 cctcaacaaa agaagtctta gacgccacct taattcatca atcaatcact ggattgtatg
     6721 agacaagaat tgatttgtct caattgggtg gtgatgaagg ggctgatcct aagaagaaaa
     6781 gaaaagttga tccaaagaaa aagcgtaagg tggatcctaa gaaaaagaga aaggttgact
     6841 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     6901 agtgataatc tcttctcgag tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     6961 ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     7021 ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt
     7081 ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     7141 tttgggacgc tcgaaggctt taatttgcgg ccggtactgc ggtgtgaaat accgcacaga
     7201 tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg
     7261 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     7321 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     7381 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     7441 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     7501 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     7561 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     7621 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     7681 cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7741 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     7801 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact
     7861 atagggcgaa ttggagctcc accgcggtgg cggccgctct agaactagtg gatcccccgg
     7921 gctgcaggaa ttcgatatca agctttcttt gaaaagataa tgtatgatta tgctttcact
     7981 catatttata cagaaacttg atgttttctt tcgagtatat acaaggtgat tacatgtacg
     8041 tttgaagtac aactctagat tttgtagtgc cctcttgggc tagcggtaaa ggtgcgcatt
     8101 ttttcacacc ctacaatgtt ctgttcaaaa gattttggtc aaacgctgta gaagtgaaag
     8161 ttggtgcgca tgtttcggcg ttcgaaactt ctccgcagtg aaagataaat gatcctagat
     8221 gcgagatatc ctcatgccta ggttttagag ctagaaatag caagttaaaa taaggctagt
     8281 ccgttatcaa cttgaaaaag tggcaccgag tcggtggtgc tttttttgtt ttttatgtct
     8341 accgtcgacc tcgagggggg gcccggtacc cagcttttgt tccctttagt gagggttaat
     8401 ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     8461 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     8521 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     8581 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     8641 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     8701 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     8761 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     8821 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     8881 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     8941 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     9001 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     9061 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     9121 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     9181 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     9241 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     9301 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     9361 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     9421 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     9481 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     9541 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     9601 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     9661 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     9721 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     9781 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     9841 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     9901 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     9961 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    10021 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    10081 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    10141 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    10201 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    10261 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    10321 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    10381 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    10441 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    10501 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    10561 aaaagtgcca cctgggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt
    10621 ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt
    10681 ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat
    10741 cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga
    10801 ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat
    10861 ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt
    10921 ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact
    10981 ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat
    11041 tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg tctaagaaac
    11101 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
