LOCUS       sequence-360674-       10566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sp.Cas9 and gRNA yeast expression vector for HDR-based gRNA 
            introduction. Selection = Nourseothricin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10566)
  AUTHORS   Bean BDM, Whiteway M, Martin VJJ
  TITLE     The MyLo CRISPR-Cas9 Toolkit: A Markerless Yeast Localization and 
            Overexpression CRISPR-Cas9 Toolkit.
  JOURNAL   G3 (Bethesda). 2022 Jun 16. pii: 6609175. doi: 
            10.1093/g3journal/jkac154.
  PUBMED    35708612
REFERENCE   2  (bases 1 to 10566)
  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: "G3 
            (Bethesda). 2022 Jun 16. pii: 6609175. doi: 
            10.1093/g3journal/jkac154."
COMMENT     Sequence Label: pBBK05 pCAS-Tyr-[gRNA: BsaI GFP dropout] (NatR...
FEATURES             Location/Qualifiers
     source          1..10566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(962..1009)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     RBS             1515..1526
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1533..2246
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2258..2329
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2345..2372
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        2386..2461
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2595..2937
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3517..3714
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(3865..4656,4657..4725))
                     /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     4488..4507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4726..4830)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5533..9636
                     /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             5533..9636
                     /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             9646..9666
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9682..9705
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      9709..9898
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(9955..10543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10035..10054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca
       61 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
      121 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt
      181 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
      241 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
      301 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
      361 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
      421 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
      481 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
      541 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
      601 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
      661 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
      721 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
      781 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
      841 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagcccg ttacagtccg
      901 gtgcgttttt ggttttttga aagtgcgtct tcagagcgct tttggttttc aaaagcgctc
      961 tgaagttcct atactttcta gagaatagga acttcggaat aggaacttca aagcgtttcc
     1021 gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt
     1081 cgcacctata tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg
     1141 tgtttatgct taaatgcgta tatgtgttat gtagtatact ctttcttcaa caattaaata
     1201 ctctcggtag ccaagttggt ttaaggcgca agactgtaat ttatcactac gaaatcttga
     1261 gatcgggcgt tcgactcgcc cccgggagag atggccggca tggtcccagc ctcctcgctg
     1321 gcgccggctg ggcaacacct tcgggtggcg aatgggactc gagaccgaaa gtgaaacgtg
     1381 atttcatgcg tcattttgaa cattttgtaa atcttattta ataatgtgtg cggcaattca
     1441 catttaattt atgaatgttt tcttaacatc gcggcaactc aagaaacggc aggttcggat
     1501 cttagctact agagaaagag gagaaatact agatgcgtaa aggcgaagag ctgttcactg
     1561 gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc
     1621 gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta
     1681 ctggtaaact gccggttcct tggccgactc tggtaacgac gctgacttat ggtgttcagt
     1741 gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg
     1801 aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg
     1861 cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact
     1921 ttaaagagga cggcaatatc ctgggccata agctggaata caattttaac agccacaatg
     1981 tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca
     2041 acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg
     2101 atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag
     2161 atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca
     2221 cgcatggtat ggatgaactg tacaaatgac caggcatcaa ataaaacgaa aggctcagtc
     2281 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     2341 cactggctca ccttcgggtg ggcctttctg cgtttatagg tctccgtttt agagctagaa
     2401 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     2461 ctttttttat tttttgtcac tattgttatg taaaatgcca cctctgacag tatggaacgc
     2521 aaacttctgt ctagtggata taggtctaga gatctgttta gcttgcctcg tccccgccgg
     2581 gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca
     2641 gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat
     2701 catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc
     2761 tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg
     2821 cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac
     2881 ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaacaat
     2941 gggtactacc ttagatgata cagcctacag atacagaaca tcagtccctg gtgatgctga
     3001 agcaattgag gctttagacg gttcattcac caccgacacc gtctttagag taaccgccac
     3061 cggtgatgga tttaccttaa gagaagtccc agtcgaccct ccattaacta aagtctttcc
     3121 agatgatgaa tctgatgacg aaagcgacga cggagaagat ggtgacccag attcaagaac
     3181 tttcgtagca tacggtgatg acggtgattt ggctggtttt gtagtcgttt cttattcagg
     3241 ttggaataga aggttgaccg ttgaagatat agaagtcgcc ccagagcata gaggtcatgg
     3301 tgtaggaaga gctttgatgg gtttggctac agaatttgca agagagagag gagccggtca
     3361 tttatggtta gaagttacta atgttaacgc ccctgctatc catgcttata gaagaatggg
     3421 tttcacatta tgtggtttag atactgcttt atatgatgga acagcatctg acggtgaaca
     3481 ggccttgtat atgtctatgc cttgccctta atcagtactg acaataaaaa gattcttgtt
     3541 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
     3601 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
     3661 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg
     3721 attcgatact aacgccgcca tccagtgtcg aaaacgagct ctcgagaacc cttaatcaaa
     3781 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3841 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3901 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3961 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgggacc cacgctcacc
     4021 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4081 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4141 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4201 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4261 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4321 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4381 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4441 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4501 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4561 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4621 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4681 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4741 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4801 ttagaaaaat aaacaaatag gggttcgtcg acagtcgaac aagaagcagg caaagtttag
     4861 agcactgccc ctccgcactc aaaaaagaaa aaactaggag gaaaataaaa ttctcaacca
     4921 cacaaacaca taaacacata caaatacaaa tacaagctta tttacttgac atcgcgcgat
     4981 cttccactat tcagcgccgt ccgccctctc tcgtgttttt tgtttacgcg acaactatgc
     5041 gaaatccgga gcaacgggca accgtttggg gaaagaccac acccacgcgc gatcgccatg
     5101 gcaacgaggt cgcacacgcc ccacacccag acctccctgc gagcgggcat gggtacaatg
     5161 tccccgttgc cacagacacc acttcgtagc acagcgcaga gcgtagcgtg ttgttgctgc
     5221 tgacaaaaga aaatttttct tagcaaagca aaggagggga agcacgggca gatagcaccg
     5281 taccataccc ttggaaactc gaaatgaacg aagcaggaaa tgagagaatg agagttttgt
     5341 aggtatatat agcggtagtg tttgcgcgtt accatcatct tctggatcta tctattgttc
     5401 ttttcctcat cactttcccc tttttcgctc ttcttcttgt cttttatttc tttctttttt
     5461 ttaattgttc cctcgattgg ctatctacca aagaatccaa acttaataca cgtatttatt
     5521 tgtccaatta ccatggataa gaaatactca ataggcttag atatcggcac aaatagcgtc
     5581 ggatgggcgg tgatcactga tgaatataag gttccgtcta aaaagttcaa ggttctggga
     5641 aatacagacc gccacagtat caaaaaaaat cttatagggg ctcttttatt tgacagtgga
     5701 gagacagcgg aagcgactcg actcaaacgg acagctcgta gaaggtatac acgtcggaag
     5761 aatcgtattt gttatctaca ggagattttt tcaaatgaga tggcgaaagt agatgatagt
     5821 ttctttcatc gacttgaaga gtcttttttg gtggaagaag acaagaagca tgaacgtcat
     5881 cctatttttg gaaatatagt agatgaagtt gcttatcatg agaaatatcc aactatctat
     5941 catctgcgaa aaaaattggt agattctact gataaagcgg atttgcgctt aatctatttg
     6001 gccttagcgc atatgattaa gtttcgtggt cattttttga ttgagggaga tttaaatcct
     6061 gataatagtg atgtggacaa actatttatc cagttggtac aaacctacaa tcaattattt
     6121 gaagaaaacc ctattaacgc aagtggagta gatgctaaag cgattctttc tgcacgattg
     6181 agtaaatcaa gacgattaga aaatctcatt gctcagctcc ccggtgagaa gaaaaatggc
     6241 ttatttggga atctcattgc tttgtcattg ggtttgaccc ctaattttaa atcaaatttt
     6301 gatttggcag aagatgctaa attacagctt tcaaaagata cttacgatga tgatttagat
     6361 aatttattgg cgcaaattgg agatcaatat gctgatttgt ttttggcagc taagaattta
     6421 tcagatgcta ttttactttc agatatccta agagtaaata ctgaaataac taaggctccc
     6481 ctatcagctt caatgattaa acgctacgat gaacatcatc aagacttgac tcttttaaaa
     6541 gctttagttc gacaacaact tccagaaaag tataaagaaa tcttttttga tcaatcaaaa
     6601 aacggatatg caggttatat tgatggggga gctagccaag aagaatttta taaatttatc
     6661 aaaccaattt tagaaaaaat ggatggtact gaggaattat tggtgaaact aaatcgtgaa
     6721 gatttgctgc gcaagcaacg gacctttgac aacggctcta ttccccatca aattcacttg
     6781 ggtgagctgc atgctatttt gagaagacaa gaagactttt atccattttt aaaagacaat
     6841 cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt attatgttgg tccattggcg
     6901 cgtggcaata gtcgttttgc atggatgact cggaagtctg aagaaacaat taccccatgg
     6961 aattttgaag aagttgtcga taaaggtgct tcagctcaat catttattga acgcatgaca
     7021 aactttgata aaaatcttcc aaatgaaaaa gtactaccaa aacatagttt gctttatgag
     7081 tattttacgg tttataacga attgacaaag gtcaaatatg ttactgaagg aatgcgaaaa
     7141 ccagcatttc tttcaggtga acagaagaaa gccattgttg atttactctt caaaacaaat
     7201 cgaaaagtaa ccgttaagca attaaaagaa gattatttca aaaaaataga atgttttgat
     7261 agtgttgaaa tttcaggagt tgaagataga tttaatgctt cattaggtac ctaccatgat
     7321 ttgctaaaaa ttattaaaga taaagatttt ttggataatg aagaaaatga agatatctta
     7381 gaggatattg ttttaacatt gaccttattt gaagataggg agatgattga ggaaagactt
     7441 aaaacatatg ctcacctctt tgatgataag gtgatgaaac agcttaaacg tcgccgttat
     7501 actggttggg gacgtttgtc tcgaaaattg attaatggta ttagggataa gcaatctggc
     7561 aaaacaatat tagatttttt gaaatcagat ggttttgcca atcgcaattt tatgcagctg
     7621 atccatgatg atagtttgac atttaaagaa gacattcaaa aagcacaagt gtctggacaa
     7681 ggcgatagtt tacatgaaca tattgcaaat ttagctggta gccctgctat taaaaaaggt
     7741 attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag taatggggcg gcataagcca
     7801 gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa ctcaaaaggg ccagaaaaat
     7861 tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag aattaggaag tcagattctt
     7921 aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa agctctatct ctattatctc
     7981 caaaatggaa gagacatgta tgtggaccaa gaattagata ttaatcgttt aagtgattat
     8041 gatgtcgatc acattgttcc acaaagtttc cttaaagacg attcaataga caataaggtc
     8101 ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg ttccaagtga agaagtagtc
     8161 aaaaagatga aaaactattg gagacaactt ctaaacgcca agttaatcac tcaacgtaag
     8221 tttgataatt taacgaaagc tgaacgtgga ggtttgagtg aacttgataa agctggtttt
     8281 atcaaacgcc aattggttga aactcgccaa atcactaagc atgtggcaca aattttggat
     8341 agtcgcatga atactaaata cgatgaaaat gataaactta ttcgagaggt taaagtgatt
     8401 accttaaaat ctaaattagt ttctgacttc cgaaaagatt tccaattcta taaagtacgt
     8461 gagattaaca attaccatca tgcccatgat gcgtatctaa atgccgtcgt tggaactgct
     8521 ttgattaaga aatatccaaa acttgaatcg gagtttgtct atggtgatta taaagtttat
     8581 gatgttcgta aaatgattgc taagtctgag caagaaatag gcaaagcaac cgcaaaatat
     8641 ttcttttact ctaatatcat gaacttcttc aaaacagaaa ttacacttgc aaatggagag
     8701 attcgcaaac gccctctaat cgaaactaat ggggaaactg gagaaattgt ctgggataaa
     8761 gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc cccaagtcaa tattgtcaag
     8821 aaaacagaag tacagacagg cggattctcc aaggagtcaa ttttaccaaa aagaaattcg
     8881 gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa aatatggtgg ttttgatagt
     8941 ccaacggtag cttattcagt cctagtggtt gctaaggtgg aaaaagggaa atcgaagaag
     9001 ttaaaatccg ttaaagagtt actagggatc acaattatgg aaagaagttc ctttgaaaaa
     9061 aatccgattg actttttaga agctaaagga tataaggaag ttaaaaaaga cttaatcatt
     9121 aaactaccta aatatagtct ttttgagtta gaaaacggtc gtaaacggat gctggctagt
     9181 gccggagaat tacaaaaagg aaatgagctg gctctgccaa gcaaatatgt gaatttttta
     9241 tatttagcta gtcattatga aaagttgaag ggtagtccag aagataacga acaaaaacaa
     9301 ttgtttgtgg agcagcataa gcattattta gatgagatta ttgagcaaat cagtgaattt
     9361 tctaagcgtg ttattttagc agatgccaat ttagataaag ttcttagtgc atataacaaa
     9421 catagagaca aaccaatacg tgaacaagca gaaaatatta ttcatttatt tacgttgacg
     9481 aatcttggag ctcccgctgc ttttaaatat tttgatacaa caattgatcg taaacgatat
     9541 acgtctacaa aagaagtttt agatgccact cttatccatc aatccatcac tggtctttat
     9601 gaaacacgca ttgatttgag tcagctagga ggtgacggtg gagggccaaa aaagaaaaga
     9661 aaagttgaag atgcttctgc tcatcatcac catcaccatc accattaaat ccgctctaac
     9721 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     9781 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     9841 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaagtg
     9901 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     9961 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
    10021 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
    10081 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
    10141 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
    10201 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
    10261 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
    10321 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
    10381 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
    10441 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
    10501 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
    10561 ttctac
//
