LOCUS       sequence-360654-       11321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sp.Cas9 and gRNA yeast expression vector for Golden Gate pre-cloning
            of gRNA. Selection = SplitKanamycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11321)
  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 11321)
  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: pBBK02 pCAS-Tyr-[gRNA: BsaI GFP dropout] (Spli...
FEATURES             Location/Qualifiers
     source          1..11321
                     /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"
     primer_bind     complement(3006..3025)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      4272..4469
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4620..5411,5412..5480))
                     /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     5243..5262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5481..5585)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6288..10391
                     /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             6288..10391
                     /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             10401..10421
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10437..10460
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      10464..10653
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(10710..11298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10790..10809)
                     /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 gggtaaggaa aagactcacg tttcgaggcc gcgattaaat tccaacatgg atgctgattt
     3001 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
     3061 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     3121 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     3181 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     3241 caaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     3301 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     3361 cgatcgcgta tttcgtctgg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3421 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3481 taagcttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3541 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcga
     3601 cgtcgctgcc tgcaggatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     3661 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     3721 cctgatgatg catggttact caccactgcg atccccggca aaacagcatt ccaggtatta
     3781 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     3841 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctggct
     3901 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     3961 aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg
     4021 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     4081 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     4141 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     4201 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     4261 ttctaatcag tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt
     4321 ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc
     4381 gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca
     4441 atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag
     4501 tgtcgaaaac gagctctcga gaacccttaa tcaaaaagga tcttcaccta gatcctttta
     4561 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4621 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4681 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4741 agtgctgcaa tgataccgcg ggacccacgc tcaccggctc cagatttatc agcaataaac
     4801 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4861 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4921 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4981 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5041 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5101 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5161 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5221 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5281 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5341 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5401 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5461 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5521 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5581 cgtcgacagt cgaacaagaa gcaggcaaag tttagagcac tgcccctccg cactcaaaaa
     5641 agaaaaaact aggaggaaaa taaaattctc aaccacacaa acacataaac acatacaaat
     5701 acaaatacaa gcttatttac ttgacatcgc gcgatcttcc actattcagc gccgtccgcc
     5761 ctctctcgtg ttttttgttt acgcgacaac tatgcgaaat ccggagcaac gggcaaccgt
     5821 ttggggaaag accacaccca cgcgcgatcg ccatggcaac gaggtcgcac acgccccaca
     5881 cccagacctc cctgcgagcg ggcatgggta caatgtcccc gttgccacag acaccacttc
     5941 gtagcacagc gcagagcgta gcgtgttgtt gctgctgaca aaagaaaatt tttcttagca
     6001 aagcaaagga ggggaagcac gggcagatag caccgtacca tacccttgga aactcgaaat
     6061 gaacgaagca ggaaatgaga gaatgagagt tttgtaggta tatatagcgg tagtgtttgc
     6121 gcgttaccat catcttctgg atctatctat tgttcttttc ctcatcactt tccccttttt
     6181 cgctcttctt cttgtctttt atttctttct tttttttaat tgttccctcg attggctatc
     6241 taccaaagaa tccaaactta atacacgtat ttatttgtcc aattaccatg gataagaaat
     6301 actcaatagg cttagatatc ggcacaaata gcgtcggatg ggcggtgatc actgatgaat
     6361 ataaggttcc gtctaaaaag ttcaaggttc tgggaaatac agaccgccac agtatcaaaa
     6421 aaaatcttat aggggctctt ttatttgaca gtggagagac agcggaagcg actcgactca
     6481 aacggacagc tcgtagaagg tatacacgtc ggaagaatcg tatttgttat ctacaggaga
     6541 ttttttcaaa tgagatggcg aaagtagatg atagtttctt tcatcgactt gaagagtctt
     6601 ttttggtgga agaagacaag aagcatgaac gtcatcctat ttttggaaat atagtagatg
     6661 aagttgctta tcatgagaaa tatccaacta tctatcatct gcgaaaaaaa ttggtagatt
     6721 ctactgataa agcggatttg cgcttaatct atttggcctt agcgcatatg attaagtttc
     6781 gtggtcattt tttgattgag ggagatttaa atcctgataa tagtgatgtg gacaaactat
     6841 ttatccagtt ggtacaaacc tacaatcaat tatttgaaga aaaccctatt aacgcaagtg
     6901 gagtagatgc taaagcgatt ctttctgcac gattgagtaa atcaagacga ttagaaaatc
     6961 tcattgctca gctccccggt gagaagaaaa atggcttatt tgggaatctc attgctttgt
     7021 cattgggttt gacccctaat tttaaatcaa attttgattt ggcagaagat gctaaattac
     7081 agctttcaaa agatacttac gatgatgatt tagataattt attggcgcaa attggagatc
     7141 aatatgctga tttgtttttg gcagctaaga atttatcaga tgctatttta ctttcagata
     7201 tcctaagagt aaatactgaa ataactaagg ctcccctatc agcttcaatg attaaacgct
     7261 acgatgaaca tcatcaagac ttgactcttt taaaagcttt agttcgacaa caacttccag
     7321 aaaagtataa agaaatcttt tttgatcaat caaaaaacgg atatgcaggt tatattgatg
     7381 ggggagctag ccaagaagaa ttttataaat ttatcaaacc aattttagaa aaaatggatg
     7441 gtactgagga attattggtg aaactaaatc gtgaagattt gctgcgcaag caacggacct
     7501 ttgacaacgg ctctattccc catcaaattc acttgggtga gctgcatgct attttgagaa
     7561 gacaagaaga cttttatcca tttttaaaag acaatcgtga gaagattgaa aaaatcttga
     7621 cttttcgaat tccttattat gttggtccat tggcgcgtgg caatagtcgt tttgcatgga
     7681 tgactcggaa gtctgaagaa acaattaccc catggaattt tgaagaagtt gtcgataaag
     7741 gtgcttcagc tcaatcattt attgaacgca tgacaaactt tgataaaaat cttccaaatg
     7801 aaaaagtact accaaaacat agtttgcttt atgagtattt tacggtttat aacgaattga
     7861 caaaggtcaa atatgttact gaaggaatgc gaaaaccagc atttctttca ggtgaacaga
     7921 agaaagccat tgttgattta ctcttcaaaa caaatcgaaa agtaaccgtt aagcaattaa
     7981 aagaagatta tttcaaaaaa atagaatgtt ttgatagtgt tgaaatttca ggagttgaag
     8041 atagatttaa tgcttcatta ggtacctacc atgatttgct aaaaattatt aaagataaag
     8101 attttttgga taatgaagaa aatgaagata tcttagagga tattgtttta acattgacct
     8161 tatttgaaga tagggagatg attgaggaaa gacttaaaac atatgctcac ctctttgatg
     8221 ataaggtgat gaaacagctt aaacgtcgcc gttatactgg ttggggacgt ttgtctcgaa
     8281 aattgattaa tggtattagg gataagcaat ctggcaaaac aatattagat tttttgaaat
     8341 cagatggttt tgccaatcgc aattttatgc agctgatcca tgatgatagt ttgacattta
     8401 aagaagacat tcaaaaagca caagtgtctg gacaaggcga tagtttacat gaacatattg
     8461 caaatttagc tggtagccct gctattaaaa aaggtatttt acagactgta aaagttgttg
     8521 atgaattggt caaagtaatg gggcggcata agccagaaaa tatcgttatt gaaatggcac
     8581 gtgaaaatca gacaactcaa aagggccaga aaaattcgcg agagcgtatg aaacgaatcg
     8641 aagaaggtat caaagaatta ggaagtcaga ttcttaaaga gcatcctgtt gaaaatactc
     8701 aattgcaaaa tgaaaagctc tatctctatt atctccaaaa tggaagagac atgtatgtgg
     8761 accaagaatt agatattaat cgtttaagtg attatgatgt cgatcacatt gttccacaaa
     8821 gtttccttaa agacgattca atagacaata aggtcttaac gcgttctgat aaaaatcgtg
     8881 gtaaatcgga taacgttcca agtgaagaag tagtcaaaaa gatgaaaaac tattggagac
     8941 aacttctaaa cgccaagtta atcactcaac gtaagtttga taatttaacg aaagctgaac
     9001 gtggaggttt gagtgaactt gataaagctg gttttatcaa acgccaattg gttgaaactc
     9061 gccaaatcac taagcatgtg gcacaaattt tggatagtcg catgaatact aaatacgatg
     9121 aaaatgataa acttattcga gaggttaaag tgattacctt aaaatctaaa ttagtttctg
     9181 acttccgaaa agatttccaa ttctataaag tacgtgagat taacaattac catcatgccc
     9241 atgatgcgta tctaaatgcc gtcgttggaa ctgctttgat taagaaatat ccaaaacttg
     9301 aatcggagtt tgtctatggt gattataaag tttatgatgt tcgtaaaatg attgctaagt
     9361 ctgagcaaga aataggcaaa gcaaccgcaa aatatttctt ttactctaat atcatgaact
     9421 tcttcaaaac agaaattaca cttgcaaatg gagagattcg caaacgccct ctaatcgaaa
     9481 ctaatgggga aactggagaa attgtctggg ataaagggcg agattttgcc acagtgcgca
     9541 aagtattgtc catgccccaa gtcaatattg tcaagaaaac agaagtacag acaggcggat
     9601 tctccaagga gtcaatttta ccaaaaagaa attcggacaa gcttattgct cgtaaaaaag
     9661 actgggatcc aaaaaaatat ggtggttttg atagtccaac ggtagcttat tcagtcctag
     9721 tggttgctaa ggtggaaaaa gggaaatcga agaagttaaa atccgttaaa gagttactag
     9781 ggatcacaat tatggaaaga agttcctttg aaaaaaatcc gattgacttt ttagaagcta
     9841 aaggatataa ggaagttaaa aaagacttaa tcattaaact acctaaatat agtctttttg
     9901 agttagaaaa cggtcgtaaa cggatgctgg ctagtgccgg agaattacaa aaaggaaatg
     9961 agctggctct gccaagcaaa tatgtgaatt ttttatattt agctagtcat tatgaaaagt
    10021 tgaagggtag tccagaagat aacgaacaaa aacaattgtt tgtggagcag cataagcatt
    10081 atttagatga gattattgag caaatcagtg aattttctaa gcgtgttatt ttagcagatg
    10141 ccaatttaga taaagttctt agtgcatata acaaacatag agacaaacca atacgtgaac
    10201 aagcagaaaa tattattcat ttatttacgt tgacgaatct tggagctccc gctgctttta
    10261 aatattttga tacaacaatt gatcgtaaac gatatacgtc tacaaaagaa gttttagatg
    10321 ccactcttat ccatcaatcc atcactggtc tttatgaaac acgcattgat ttgagtcagc
    10381 taggaggtga cggtggaggg ccaaaaaaga aaagaaaagt tgaagatgct tctgctcatc
    10441 atcaccatca ccatcaccat taaatccgct ctaaccgaaa aggaaggagt tagacaacct
    10501 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
    10561 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
    10621 accttgcttg agaaggtttt gggacgctcg aagtgagcaa aaggccagca aaaggccagg
    10681 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
    10741 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
    10801 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
    10861 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
    10921 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
    10981 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
    11041 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
    11101 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
    11161 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
    11221 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
    11281 agaaaaaaag gatctcaaga agatcctttg atcttttcta c
//
