LOCUS       sequence-360673-       11532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sp.Cas9 and gRNA yeast expression vector for Golden Gate pre-cloning
            of gRNA. Selection = SplitHygromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11532)
  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 11532)
  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: pBBK04 pCAS-Tyr-[gRNA: BsaI GFP dropout]
            (Spli...
FEATURES             Location/Qualifiers
     source          1..11532
                     /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      4483..4680
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4831..5622,5623..5691))
                     /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     5454..5473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5692..5796)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6499..10602
                     /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             6499..10602
                     /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             10612..10632
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10648..10671
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      10675..10864
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(10921..11509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11001..11020)
                     /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 gggaaagaaa ccagaattga cagctacgtc tgtcgagaag tttctgatcg aaaagttcga
     3001 cagcgtgtcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     3061 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     3121 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     3181 tggggaattt agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     3241 gcaagacctg cctgaaaccg aactgcccgc tgttctgcaa ccggtcgcgg aggccatgga
     3301 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     3361 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     3421 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     3481 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     3541 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     3601 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     3661 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgac
     3721 gtcgctgcct gcaggacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     3781 tgttctgcaa ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac
     3841 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     3901 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     3961 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     4021 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     4081 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     4141 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4201 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
     4261 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4321 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     4381 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4441 ccgacgcccc agcactcgtc cgagggcaaa ggaataatca gtactgacaa taaaaagatt
     4501 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     4561 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     4621 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     4681 ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa cgagctctcg agaaccctta
     4741 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4801 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4861 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4921 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gggacccacg
     4981 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5041 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5101 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5161 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5221 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5281 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5341 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5401 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5461 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5521 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5581 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5641 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5701 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5761 atgtatttag aaaaataaac aaataggggt tcgtcgacag tcgaacaaga agcaggcaaa
     5821 gtttagagca ctgcccctcc gcactcaaaa aagaaaaaac taggaggaaa ataaaattct
     5881 caaccacaca aacacataaa cacatacaaa tacaaataca agcttattta cttgacatcg
     5941 cgcgatcttc cactattcag cgccgtccgc cctctctcgt gttttttgtt tacgcgacaa
     6001 ctatgcgaaa tccggagcaa cgggcaaccg tttggggaaa gaccacaccc acgcgcgatc
     6061 gccatggcaa cgaggtcgca cacgccccac acccagacct ccctgcgagc gggcatgggt
     6121 acaatgtccc cgttgccaca gacaccactt cgtagcacag cgcagagcgt agcgtgttgt
     6181 tgctgctgac aaaagaaaat ttttcttagc aaagcaaagg aggggaagca cgggcagata
     6241 gcaccgtacc atacccttgg aaactcgaaa tgaacgaagc aggaaatgag agaatgagag
     6301 ttttgtaggt atatatagcg gtagtgtttg cgcgttacca tcatcttctg gatctatcta
     6361 ttgttctttt cctcatcact ttcccctttt tcgctcttct tcttgtcttt tatttctttc
     6421 ttttttttaa ttgttccctc gattggctat ctaccaaaga atccaaactt aatacacgta
     6481 tttatttgtc caattaccat ggataagaaa tactcaatag gcttagatat cggcacaaat
     6541 agcgtcggat gggcggtgat cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt
     6601 ctgggaaata cagaccgcca cagtatcaaa aaaaatctta taggggctct tttatttgac
     6661 agtggagaga cagcggaagc gactcgactc aaacggacag ctcgtagaag gtatacacgt
     6721 cggaagaatc gtatttgtta tctacaggag attttttcaa atgagatggc gaaagtagat
     6781 gatagtttct ttcatcgact tgaagagtct tttttggtgg aagaagacaa gaagcatgaa
     6841 cgtcatccta tttttggaaa tatagtagat gaagttgctt atcatgagaa atatccaact
     6901 atctatcatc tgcgaaaaaa attggtagat tctactgata aagcggattt gcgcttaatc
     6961 tatttggcct tagcgcatat gattaagttt cgtggtcatt ttttgattga gggagattta
     7021 aatcctgata atagtgatgt ggacaaacta tttatccagt tggtacaaac ctacaatcaa
     7081 ttatttgaag aaaaccctat taacgcaagt ggagtagatg ctaaagcgat tctttctgca
     7141 cgattgagta aatcaagacg attagaaaat ctcattgctc agctccccgg tgagaagaaa
     7201 aatggcttat ttgggaatct cattgctttg tcattgggtt tgacccctaa ttttaaatca
     7261 aattttgatt tggcagaaga tgctaaatta cagctttcaa aagatactta cgatgatgat
     7321 ttagataatt tattggcgca aattggagat caatatgctg atttgttttt ggcagctaag
     7381 aatttatcag atgctatttt actttcagat atcctaagag taaatactga aataactaag
     7441 gctcccctat cagcttcaat gattaaacgc tacgatgaac atcatcaaga cttgactctt
     7501 ttaaaagctt tagttcgaca acaacttcca gaaaagtata aagaaatctt ttttgatcaa
     7561 tcaaaaaacg gatatgcagg ttatattgat gggggagcta gccaagaaga attttataaa
     7621 tttatcaaac caattttaga aaaaatggat ggtactgagg aattattggt gaaactaaat
     7681 cgtgaagatt tgctgcgcaa gcaacggacc tttgacaacg gctctattcc ccatcaaatt
     7741 cacttgggtg agctgcatgc tattttgaga agacaagaag acttttatcc atttttaaaa
     7801 gacaatcgtg agaagattga aaaaatcttg acttttcgaa ttccttatta tgttggtcca
     7861 ttggcgcgtg gcaatagtcg ttttgcatgg atgactcgga agtctgaaga aacaattacc
     7921 ccatggaatt ttgaagaagt tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc
     7981 atgacaaact ttgataaaaa tcttccaaat gaaaaagtac taccaaaaca tagtttgctt
     8041 tatgagtatt ttacggttta taacgaattg acaaaggtca aatatgttac tgaaggaatg
     8101 cgaaaaccag catttctttc aggtgaacag aagaaagcca ttgttgattt actcttcaaa
     8161 acaaatcgaa aagtaaccgt taagcaatta aaagaagatt atttcaaaaa aatagaatgt
     8221 tttgatagtg ttgaaatttc aggagttgaa gatagattta atgcttcatt aggtacctac
     8281 catgatttgc taaaaattat taaagataaa gattttttgg ataatgaaga aaatgaagat
     8341 atcttagagg atattgtttt aacattgacc ttatttgaag atagggagat gattgaggaa
     8401 agacttaaaa catatgctca cctctttgat gataaggtga tgaaacagct taaacgtcgc
     8461 cgttatactg gttggggacg tttgtctcga aaattgatta atggtattag ggataagcaa
     8521 tctggcaaaa caatattaga ttttttgaaa tcagatggtt ttgccaatcg caattttatg
     8581 cagctgatcc atgatgatag tttgacattt aaagaagaca ttcaaaaagc acaagtgtct
     8641 ggacaaggcg atagtttaca tgaacatatt gcaaatttag ctggtagccc tgctattaaa
     8701 aaaggtattt tacagactgt aaaagttgtt gatgaattgg tcaaagtaat ggggcggcat
     8761 aagccagaaa atatcgttat tgaaatggca cgtgaaaatc agacaactca aaagggccag
     8821 aaaaattcgc gagagcgtat gaaacgaatc gaagaaggta tcaaagaatt aggaagtcag
     8881 attcttaaag agcatcctgt tgaaaatact caattgcaaa atgaaaagct ctatctctat
     8941 tatctccaaa atggaagaga catgtatgtg gaccaagaat tagatattaa tcgtttaagt
     9001 gattatgatg tcgatcacat tgttccacaa agtttcctta aagacgattc aatagacaat
     9061 aaggtcttaa cgcgttctga taaaaatcgt ggtaaatcgg ataacgttcc aagtgaagaa
     9121 gtagtcaaaa agatgaaaaa ctattggaga caacttctaa acgccaagtt aatcactcaa
     9181 cgtaagtttg ataatttaac gaaagctgaa cgtggaggtt tgagtgaact tgataaagct
     9241 ggttttatca aacgccaatt ggttgaaact cgccaaatca ctaagcatgt ggcacaaatt
     9301 ttggatagtc gcatgaatac taaatacgat gaaaatgata aacttattcg agaggttaaa
     9361 gtgattacct taaaatctaa attagtttct gacttccgaa aagatttcca attctataaa
     9421 gtacgtgaga ttaacaatta ccatcatgcc catgatgcgt atctaaatgc cgtcgttgga
     9481 actgctttga ttaagaaata tccaaaactt gaatcggagt ttgtctatgg tgattataaa
     9541 gtttatgatg ttcgtaaaat gattgctaag tctgagcaag aaataggcaa agcaaccgca
     9601 aaatatttct tttactctaa tatcatgaac ttcttcaaaa cagaaattac acttgcaaat
     9661 ggagagattc gcaaacgccc tctaatcgaa actaatgggg aaactggaga aattgtctgg
     9721 gataaagggc gagattttgc cacagtgcgc aaagtattgt ccatgcccca agtcaatatt
     9781 gtcaagaaaa cagaagtaca gacaggcgga ttctccaagg agtcaatttt accaaaaaga
     9841 aattcggaca agcttattgc tcgtaaaaaa gactgggatc caaaaaaata tggtggtttt
     9901 gatagtccaa cggtagctta ttcagtccta gtggttgcta aggtggaaaa agggaaatcg
     9961 aagaagttaa aatccgttaa agagttacta gggatcacaa ttatggaaag aagttccttt
    10021 gaaaaaaatc cgattgactt tttagaagct aaaggatata aggaagttaa aaaagactta
    10081 atcattaaac tacctaaata tagtcttttt gagttagaaa acggtcgtaa acggatgctg
    10141 gctagtgccg gagaattaca aaaaggaaat gagctggctc tgccaagcaa atatgtgaat
    10201 tttttatatt tagctagtca ttatgaaaag ttgaagggta gtccagaaga taacgaacaa
    10261 aaacaattgt ttgtggagca gcataagcat tatttagatg agattattga gcaaatcagt
    10321 gaattttcta agcgtgttat tttagcagat gccaatttag ataaagttct tagtgcatat
    10381 aacaaacata gagacaaacc aatacgtgaa caagcagaaa atattattca tttatttacg
    10441 ttgacgaatc ttggagctcc cgctgctttt aaatattttg atacaacaat tgatcgtaaa
    10501 cgatatacgt ctacaaaaga agttttagat gccactctta tccatcaatc catcactggt
    10561 ctttatgaaa cacgcattga tttgagtcag ctaggaggtg acggtggagg gccaaaaaag
    10621 aaaagaaaag ttgaagatgc ttctgctcat catcaccatc accatcacca ttaaatccgc
    10681 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
    10741 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag
    10801 acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc
    10861 gaagtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
    10921 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
    10981 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
    11041 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
    11101 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
    11161 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
    11221 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
    11281 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
    11341 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
    11401 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
    11461 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
    11521 gatcttttct ac
//
