LOCUS       sequence-360653-       10803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sp.Cas9 and gRNA yeast expression vector for HDR-based gRNA 
            introduction. Selection = Kanamycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10803)
  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 10803)
  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: pBBK01 pCAS-Tyr-[gRNA: BsaI GFP dropout] (KanR...
FEATURES             Location/Qualifiers
     source          1..10803
                     /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"
     gene            2595..3951
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2595..2937
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2939..3748
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3006..3025)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3754..3951
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4102..4893,4894..4962))
                     /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     4725..4744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4963..5067)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5770..9873
                     /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             5770..9873
                     /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             9883..9903
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9919..9942
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      9946..10135
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(10192..10780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10272..10291)
                     /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 tcggaatcgc
     3601 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3661 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3721 tcatttgatg ctcgatgagt ttttctaatc agtactgaca ataaaaagat tcttgttttc
     3781 aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt
     3841 agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg
     3901 cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt
     3961 cgatactaac gccgccatcc agtgtcgaaa acgagctctc gagaaccctt aatcaaaaag
     4021 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4081 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4141 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4201 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgggacccac gctcaccggc
     4261 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4321 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4381 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4441 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4501 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4561 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4621 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4681 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4741 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4801 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4861 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4921 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4981 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5041 gaaaaataaa caaatagggg ttcgtcgaca gtcgaacaag aagcaggcaa agtttagagc
     5101 actgcccctc cgcactcaaa aaagaaaaaa ctaggaggaa aataaaattc tcaaccacac
     5161 aaacacataa acacatacaa atacaaatac aagcttattt acttgacatc gcgcgatctt
     5221 ccactattca gcgccgtccg ccctctctcg tgttttttgt ttacgcgaca actatgcgaa
     5281 atccggagca acgggcaacc gtttggggaa agaccacacc cacgcgcgat cgccatggca
     5341 acgaggtcgc acacgcccca cacccagacc tccctgcgag cgggcatggg tacaatgtcc
     5401 ccgttgccac agacaccact tcgtagcaca gcgcagagcg tagcgtgttg ttgctgctga
     5461 caaaagaaaa tttttcttag caaagcaaag gaggggaagc acgggcagat agcaccgtac
     5521 catacccttg gaaactcgaa atgaacgaag caggaaatga gagaatgaga gttttgtagg
     5581 tatatatagc ggtagtgttt gcgcgttacc atcatcttct ggatctatct attgttcttt
     5641 tcctcatcac tttccccttt ttcgctcttc ttcttgtctt ttatttcttt ctttttttta
     5701 attgttccct cgattggcta tctaccaaag aatccaaact taatacacgt atttatttgt
     5761 ccaattacca tggataagaa atactcaata ggcttagata tcggcacaaa tagcgtcgga
     5821 tgggcggtga tcactgatga atataaggtt ccgtctaaaa agttcaaggt tctgggaaat
     5881 acagaccgcc acagtatcaa aaaaaatctt ataggggctc ttttatttga cagtggagag
     5941 acagcggaag cgactcgact caaacggaca gctcgtagaa ggtatacacg tcggaagaat
     6001 cgtatttgtt atctacagga gattttttca aatgagatgg cgaaagtaga tgatagtttc
     6061 tttcatcgac ttgaagagtc ttttttggtg gaagaagaca agaagcatga acgtcatcct
     6121 atttttggaa atatagtaga tgaagttgct tatcatgaga aatatccaac tatctatcat
     6181 ctgcgaaaaa aattggtaga ttctactgat aaagcggatt tgcgcttaat ctatttggcc
     6241 ttagcgcata tgattaagtt tcgtggtcat tttttgattg agggagattt aaatcctgat
     6301 aatagtgatg tggacaaact atttatccag ttggtacaaa cctacaatca attatttgaa
     6361 gaaaacccta ttaacgcaag tggagtagat gctaaagcga ttctttctgc acgattgagt
     6421 aaatcaagac gattagaaaa tctcattgct cagctccccg gtgagaagaa aaatggctta
     6481 tttgggaatc tcattgcttt gtcattgggt ttgaccccta attttaaatc aaattttgat
     6541 ttggcagaag atgctaaatt acagctttca aaagatactt acgatgatga tttagataat
     6601 ttattggcgc aaattggaga tcaatatgct gatttgtttt tggcagctaa gaatttatca
     6661 gatgctattt tactttcaga tatcctaaga gtaaatactg aaataactaa ggctccccta
     6721 tcagcttcaa tgattaaacg ctacgatgaa catcatcaag acttgactct tttaaaagct
     6781 ttagttcgac aacaacttcc agaaaagtat aaagaaatct tttttgatca atcaaaaaac
     6841 ggatatgcag gttatattga tgggggagct agccaagaag aattttataa atttatcaaa
     6901 ccaattttag aaaaaatgga tggtactgag gaattattgg tgaaactaaa tcgtgaagat
     6961 ttgctgcgca agcaacggac ctttgacaac ggctctattc cccatcaaat tcacttgggt
     7021 gagctgcatg ctattttgag aagacaagaa gacttttatc catttttaaa agacaatcgt
     7081 gagaagattg aaaaaatctt gacttttcga attccttatt atgttggtcc attggcgcgt
     7141 ggcaatagtc gttttgcatg gatgactcgg aagtctgaag aaacaattac cccatggaat
     7201 tttgaagaag ttgtcgataa aggtgcttca gctcaatcat ttattgaacg catgacaaac
     7261 tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac atagtttgct ttatgagtat
     7321 tttacggttt ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca
     7381 gcatttcttt caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga
     7441 aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt
     7501 gttgaaattt caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg
     7561 ctaaaaatta ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag
     7621 gatattgttt taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa
     7681 acatatgctc acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact
     7741 ggttggggac gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa
     7801 acaatattag attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc
     7861 catgatgata gtttgacatt taaagaagac attcaaaaag cacaagtgtc tggacaaggc
     7921 gatagtttac atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt
     7981 ttacagactg taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa
     8041 aatatcgtta ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg
     8101 cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa
     8161 gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa
     8221 aatggaagag acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat
     8281 gtcgatcaca ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta
     8341 acgcgttctg ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa
     8401 aagatgaaaa actattggag acaacttcta aacgccaagt taatcactca acgtaagttt
     8461 gataatttaa cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc
     8521 aaacgccaat tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt
     8581 cgcatgaata ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc
     8641 ttaaaatcta aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag
     8701 attaacaatt accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg
     8761 attaagaaat atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat
     8821 gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc
     8881 ttttactcta atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt
     8941 cgcaaacgcc ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg
     9001 cgagattttg ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa
     9061 acagaagtac agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac
     9121 aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca
     9181 acggtagctt attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta
     9241 aaatccgtta aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat
     9301 ccgattgact ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa
     9361 ctacctaaat atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc
     9421 ggagaattac aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat
     9481 ttagctagtc attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg
     9541 tttgtggagc agcataagca ttatttagat gagattattg agcaaatcag tgaattttct
     9601 aagcgtgtta ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat
     9661 agagacaaac caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat
     9721 cttggagctc ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg
     9781 tctacaaaag aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa
     9841 acacgcattg atttgagtca gctaggaggt gacggtggag ggccaaaaaa gaaaagaaaa
     9901 gttgaagatg cttctgctca tcatcaccat caccatcacc attaaatccg ctctaaccga
     9961 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt
    10021 agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta
    10081 cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct cgaagtgagc
    10141 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
    10201 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
    10261 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
    10321 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
    10381 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
    10441 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
    10501 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
    10561 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
    10621 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
    10681 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
    10741 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
    10801 tac
//
