LOCUS       sequence-99574-e       10956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding iCas9 and tracrRNA on pRS415 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10956)
  AUTHORS   Bao Z, Xiao H, Liang J, Zhang L, Xiong X, Sun N, Si T, Zhao H
  TITLE     Homology-Integrated CRISPR-Cas (HI-CRISPR) System for One-Step 
            Multigene Disruption in Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2014 Sep 19.
  PUBMED    25207793
REFERENCE   2  (bases 1 to 10956)
  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: "ACS Synth 
            Biol. 2014 Sep 19."
COMMENT     Sequence Label: pCT
FEATURES             Location/Qualifiers
     source          1..10956
                     /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"
     CDS             complement(663..1757)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(2611..2632)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2702..3100
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3112..3135
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3139..3159
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3160..7260
                     /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
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTYKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSITHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7264..7284
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7288..7587
                     /label=ADH2 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 2 (ADH2) gene"
     promoter        7600..8006
                     /gene="RPR1"
                     /label=RPR1 promoter
                     /note="promoter for the S. cerevisiae RNase P RNA gene"
     misc_RNA        8007..8085
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     terminator      8093..8155
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
     primer_bind     complement(8182..8202)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8182..8200)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8221..8237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8221..8237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8234..8256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8245..8261
                     /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(8269..8275,8276..8293,8294..8299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8314..8335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8452..8469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8623..9211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8703..8722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9382..10173,10174..10242))
                     /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     10005..10024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10243..10347)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    10384..10887
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     10929..10947
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aggcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gactcggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
      241 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
      301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
      361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
      421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
      481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
      541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
      601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
      661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
      721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
      781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
      841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
      901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
      961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
     1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
     1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
     1141 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata
     1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
     1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
     1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
     1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
     1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
     1501 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg
     1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
     1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
     1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
     1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
     1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
     1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
     1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
     1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
     2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
     2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
     2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
     2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
     2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
     2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
     2401 gaagttctcc tcgaccgtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2701 gcatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg
     2761 catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc
     2821 tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg
     2881 tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa
     2941 attttttttt tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac
     3001 ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact
     3061 tcttgctcat tagaaagaaa gcatagcaat ctaatctaag ttttctagat ggattataaa
     3121 gatgacgatg acaaacctcc aaaaaagaag agaaaggtcg ataagaaata ctcaataggc
     3181 ttagatatcg gcacaaatag cgtcggatgg gcggtgatca ctgatgaata taaggttccg
     3241 tctaaaaagt tcaaggttct gggaaataca gaccgccaca gtatcaaaaa aaatcttata
     3301 ggggctcttt tatttgacag tggagagaca gcggaagcga ctcgtctcaa acggacagct
     3361 cgtagaaggt atacacgtcg gaagaatcgt atttgttatc tacaggagat tttttcaaat
     3421 gagatggcga aagtagatga tagtttcttt catcgacttg aagagtcttt tttggtggaa
     3481 gaagacaaga agcatgaacg tcatcctatt tttggaaata tagtagatga agttgcttat
     3541 catgagaaat atccaactat ctatcatctg cgaaaaaaat tggtagattc tacttataaa
     3601 gcggatttgc gcttaatcta tttggcctta gcgcatatga ttaagtttcg tggtcatttt
     3661 ttgattgagg gagatttaaa tcctgataat agtgatgtgg acaaactatt tatccagttg
     3721 gtacaaacct acaatcaatt atttgaagaa aaccctatta acgcaagtgg agtagatgct
     3781 aaagcgattc tttctgcacg attgagtaaa tcaagacgat tagaaaatct cattgctcag
     3841 ctccccggtg agaagaaaaa tggcttattt gggaatctca ttgctttgtc attgggtttg
     3901 acccctaatt ttaaatcaaa ttttgatttg gcagaagatg ctaaattaca gctttcaaaa
     3961 gatacttacg atgatgattt agataattta ttggcgcaaa ttggagatca atatgctgat
     4021 ttgtttttgg cagctaagaa tttatcagat gctattttac tttcagatat cctaagagta
     4081 aatactgaaa taactaaggc tcccctatca gcttcaatga ttaaacgcta cgatgaacat
     4141 catcaagact tgactctttt aaaagcttta gttcgacaac aacttccaga aaagtataaa
     4201 gaaatctttt ttgatcaatc aaaaaacgga tatgcaggtt atattgatgg gggagctagc
     4261 caagaagaat tttataaatt tatcaaacca attttagaaa aaatggatgg tactgaggaa
     4321 ttattggtga aactaaatcg tgaagatttg ctgcgcaagc aacggacctt tgacaacggc
     4381 tctattaccc atcaaattca cttgggtgag ctgcatgcta ttttgagaag acaagaagac
     4441 ttttatccat ttttaaaaga caatcgtgag aagattgaaa aaatcttgac ttttcgaatt
     4501 ccttattatg ttggtccatt ggcgcgtggc aatagtcgtt ttgcatggat gactcggaag
     4561 tctgaagaaa caattacccc atggaatttt gaagaagttg tcgataaagg tgcttcagct
     4621 caatcattta ttgaacgcat gacaaacttt gataaaaatc ttccaaatga aaaagtacta
     4681 ccaaaacata gtttgcttta tgagtatttt acggtttata acgaattgac aaaggtcaaa
     4741 tatgttactg aaggaatgcg aaaaccagca tttctttcag gtgaacagaa gaaagccatt
     4801 gttgatttac tcttcaaaac aaatcgaaaa gtaaccgtta agcaattaaa agaagattat
     4861 ttcaaaaaaa tagaatgttt tgatagtgtt gaaatttcag gagttgaaga tagatttaat
     4921 gcttcattag gtacctacca tgatttgcta aaaattatta aagataaaga ttttttggat
     4981 aatgaagaaa atgaagatat cttagaggat attgttttaa cattgacctt atttgaagat
     5041 agggagatga ttgaggaaag acttaaaaca tatgctcacc tctttgatga taaggtgatg
     5101 aaacagctta aacgtcgccg ttatactggt tggggacgtt tgtctcgaaa attgattaat
     5161 ggtattaggg ataagcaatc tggcaaaaca atattagatt ttttgaaatc agatggtttt
     5221 gccaatcgca attttatgca gctgatccat gatgatagtt tgacatttaa agaagacatt
     5281 caaaaagcac aagtgtctgg acaaggcgat agtttacatg aacatattgc aaatttagct
     5341 ggtagccctg ctattaaaaa aggtatttta cagactgtaa aagttgttga tgaattggtc
     5401 aaagtaatgg ggcggcataa gccagaaaat atcgttattg aaatggcacg tgaaaatcag
     5461 acaactcaaa agggccagaa aaattcgcga gagcgtatga aacgaatcga agaaggtatc
     5521 aaagaattag gaagtcagat tcttaaagag catcctgttg aaaatactca attgcaaaat
     5581 gaaaagctct atctctatta tctccaaaat ggaagagaca tgtatgtgga ccaagaatta
     5641 gatattaatc gtttaagtga ttatgatgtc gatcacattg ttccacaaag tttccttaaa
     5701 gacgattcaa tagacaataa ggtcttaacg cgttctgata aaaatcgtgg taaatcggat
     5761 aacgttccaa gtgaagaagt agtcaaaaag atgaaaaact attggagaca acttctaaac
     5821 gccaagttaa tcactcaacg taagtttgat aatttaacga aagctgaacg tggaggtttg
     5881 agtgaacttg ataaagctgg ttttatcaaa cgccaattgg ttgaaactcg ccaaatcact
     5941 aagcatgtgg cacaaatttt ggatagtcgc atgaatacta aatacgatga aaatgataaa
     6001 cttattcgag aggttaaagt gattacctta aaatctaaat tagtttctga cttccgaaaa
     6061 gatttccaat tctataaagt acgtgagatt aacaattacc atcatgccca tgatgcgtat
     6121 ctaaatgccg tcgttggaac tgctttgatt aagaaatatc caaaacttga atcggagttt
     6181 gtctatggtg attataaagt ttatgatgtt cgtaaaatga ttgctaagtc tgagcaagaa
     6241 ataggcaaag caaccgcaaa atatttcttt tactctaata tcatgaactt cttcaaaaca
     6301 gaaattacac ttgcaaatgg agagattcgc aaacgccctc taatcgaaac taatggggaa
     6361 actggagaaa ttgtctggga taaagggcga gattttgcca cagtgcgcaa agtattgtcc
     6421 atgccccaag tcaatattgt caagaaaaca gaagtacaga caggcggatt ctccaaggag
     6481 tcaattttac caaaaagaaa ttcggacaag cttattgctc gtaaaaaaga ctgggatcca
     6541 aaaaaatatg gtggttttga tagtccaacg gtagcttatt cagtcctagt ggttgctaag
     6601 gtggaaaaag ggaaatcgaa gaagttaaaa tccgttaaag agttactagg gatcacaatt
     6661 atggaaagaa gttcctttga aaaaaatccg attgactttt tagaagctaa aggatataag
     6721 gaagttaaaa aagacttaat cattaaacta cctaaatata gtctttttga gttagaaaac
     6781 ggtcgtaaac ggatgctggc tagtgccgga gaattacaaa aaggaaatga gctggctctg
     6841 ccaagcaaat atgtgaattt tttatattta gctagtcatt atgaaaagtt gaagggtagt
     6901 ccagaagata acgaacaaaa acaattgttt gtggagcagc ataagcatta tttagatgag
     6961 attattgagc aaatcagtga attttctaag cgtgttattt tagcagatgc caatttagat
     7021 aaagttctta gtgcatataa caaacataga gacaaaccaa tacgtgaaca agcagaaaat
     7081 attattcatt tatttacgtt gacgaatctt ggagctcccg ctgcttttaa atattttgat
     7141 acaacaattg atcgtaaacg atatacgtct acaaaagaag ttttagatgc cactcttatc
     7201 catcaatcca tcactggtct ttatgaaaca cgcattgatt tgagtcagct aggaggtgac
     7261 cctccaaaaa agaagagaaa ggtctgagcg gatctcttat gtctttacga tttatagttt
     7321 tcattatcaa gtatgcctat attagtatat agcatcttta gatgacagtg ttcgaagttt
     7381 cacgaataaa agataatatt ctactttttg ctcccaccgc gtttgctagc acgagtgaac
     7441 accatccctc gcctgtgagt tgtacccatt cctctaaact gtagacatgg tagcttcagc
     7501 agtgttcgtt atgtacggca tcctccaaca aacagtcggt tatagtttgt cctgctcctc
     7561 tgaatcgagt ccctcgatat ttctcatact agttctagag atctgccaat tgaacataac
     7621 atggtagtta catatactag taatatggtt cggcacacat taaaagtata aaaactatct
     7681 gaattacgaa ttacatatat tggtcataaa aatcaatcaa tcatcgtgtg ttttatatgt
     7741 ctcttatcta agtataagaa tatccatagt taatattcac ttacgctacc ttttaacctg
     7801 taatcattgt caacaggata tgttaacgac ccacattgat aaacgctagt atttcttttt
     7861 cctcttctta ttggccggct gtctctatac tcccctatag tctgtttctt ttcgtttcga
     7921 ttgttttacg tttgaggcct cgtggcgcac atggtacgct gtggtgctcg cggctgggaa
     7981 cgaaactctg ggagctgcga ttggcaggaa ccattcaaaa cagcatagca agttaaaata
     8041 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttccatatcc
     8101 aacttccaat ttaatctttc ttttttaatt ttcacttatt tgcgatacag aaagaccgcg
     8161 gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     8221 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     8281 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     8341 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     8401 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     8461 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     8521 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     8581 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     8641 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     8701 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     8761 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     8821 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     8881 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     8941 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     9001 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     9061 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     9121 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     9181 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     9241 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     9301 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     9361 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     9421 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     9481 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     9541 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     9601 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     9661 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     9721 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     9781 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     9841 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9901 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9961 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
    10021 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
    10081 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
    10141 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
    10201 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
    10261 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
    10321 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcctt
    10381 ttcatcacgt gctataaaaa taattataat ttaaattttt taatataaat atataaatta
    10441 aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa
    10501 aagactctag ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg
    10561 tattaatgcc gaattgtttc atcttgtctg tgtagaagac cacacacgaa aatcctgtga
    10621 ttttacattt tacttatcgt taatcgaatg tatatctatt taatctgctt ttcttgtcta
    10681 ataaatatat atgtaaagta cgctttttgt tgaaattttt taaacctttg tttatttttt
    10741 tttcttcatt ccgtaactct tctaccttct ttatttactt tctaaaatcc aaatacaaaa
    10801 cataaaaata aataaacaca gagtaaattc ccaaattatt ccatcattaa aagatacgag
    10861 gcgcgtgtaa gttacaggca agcgatccgt cctaagaaac cattattatc atgacattaa
    10921 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
