LOCUS       sequence-360677-       11606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sp.Cas9 and gRNA yeast expression vector for Golden Gate pre-cloning
            of gRNA. Selection = SplitLEU2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11606)
  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 11606)
  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: pBBK08 pCAS-Tyr-[gRNA: BsaI GFP dropout]
            (Spli...
FEATURES             Location/Qualifiers
     source          1..11606
                     /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      4557..4754
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4905..5696,5697..5765))
                     /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     5528..5547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5766..5870)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6573..10676
                     /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             6573..10676
                     /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             10686..10706
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10722..10745
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      10749..10938
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(10995..11583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11075..11094)
                     /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 gtctgcccct aagaagatcg tcgttttgcc aggtgaccac gttggtcaag aaatcacagc
     3001 cgaagccatt aaggttctta aagctatttc tgatgttcgt tccaatgtca agttcgattt
     3061 cgaaaatcat ttaattggtg gtgctgctat cgatgctaca ggtgttccac ttccagatga
     3121 ggcgctggaa gcctccaaga aggctgatgc cgttttgtta ggtgctgtgg gtggtcctaa
     3181 atggggaacc ggtagtgtta gacctgaaca aggtttacta aaaatccgta aagaacttca
     3241 attgtacgcc aacttaagac catgtaactt tgcatccgac tctcttttag acttatctcc
     3301 aatcaagcca caatttgcta aaggtactga cttcgttgtt gtcagagaat tagtgggagg
     3361 tatttacttt ggtaagagaa aggaggacga tggtgatggt gtcgcttggg atagtgaaca
     3421 atacaccgtt ccagaagtgc aaagaatcac aagaatggcc gctttcatgg ccctacaaca
     3481 tgagccacca ttgcctattt ggtccttgga taaagctaat gttttggcct cttcaagatt
     3541 atggagaaaa actgtggagg aaaccatcaa gaacgaattt cctacattga aggttcaaca
     3601 tcaattgatt gattctgccg ccatgatcct agttaagaac ccaacccacc taaatggtat
     3661 tataatcacc agcagacgtc gctgcctgca ggtcctaaat ggggaaccgg tagtgttaga
     3721 cctgaacaag gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa cttaagacca
     3781 tgtaactttg catccgactc tcttttagac ttatctccaa tcaagccaca atttgctaaa
     3841 ggtactgact tcgttgttgt cagagaatta gtgggaggta tttactttgg taagagaaag
     3901 gaggacgatg gtgatggtgt cgcttgggat agtgaacaat acaccgttcc agaagtgcaa
     3961 agaatcacaa gaatggccgc tttcatggcc ctacaacatg agccaccatt gcctatttgg
     4021 tccttggata aagctaatgt tttggcctct tcaagattat ggagaaaaac tgtggaggaa
     4081 accatcaaga acgaatttcc tacattgaag gttcaacatc aattgattga ttctgccgcc
     4141 atgatcctag ttaagaaccc aacccaccta aatggtatta taatcaccag caacatgttt
     4201 ggtgatatca tctccgatga agcctccgtt atcccaggtt ccttgggttt gttgccatct
     4261 gcgtccttgg cctctttgcc agacaagaac accgcatttg gtttgtacga accatgccac
     4321 ggttctgctc cagatttgcc aaagaataag gtcaacccta tcgccactat cttgtctgct
     4381 gcaatgatgt tgaaattgtc attgaacttg cctgaagaag gtaaggccat tgaagatgca
     4441 gttaaaaagg ttttggatgc aggtatcaga actggtgatt taggtggttc caacagtacc
     4501 accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta atcagtactg
     4561 acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag
     4621 ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct
     4681 gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc
     4741 tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtgtcg aaaacgagct
     4801 ctcgagaacc cttaatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4861 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4921 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4981 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5041 ccgcgggacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5101 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5161 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5221 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5281 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5341 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5401 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5461 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5521 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5581 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5641 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5701 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5761 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5821 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttcgtcg acagtcgaac
     5881 aagaagcagg caaagtttag agcactgccc ctccgcactc aaaaaagaaa aaactaggag
     5941 gaaaataaaa ttctcaacca cacaaacaca taaacacata caaatacaaa tacaagctta
     6001 tttacttgac atcgcgcgat cttccactat tcagcgccgt ccgccctctc tcgtgttttt
     6061 tgtttacgcg acaactatgc gaaatccgga gcaacgggca accgtttggg gaaagaccac
     6121 acccacgcgc gatcgccatg gcaacgaggt cgcacacgcc ccacacccag acctccctgc
     6181 gagcgggcat gggtacaatg tccccgttgc cacagacacc acttcgtagc acagcgcaga
     6241 gcgtagcgtg ttgttgctgc tgacaaaaga aaatttttct tagcaaagca aaggagggga
     6301 agcacgggca gatagcaccg taccataccc ttggaaactc gaaatgaacg aagcaggaaa
     6361 tgagagaatg agagttttgt aggtatatat agcggtagtg tttgcgcgtt accatcatct
     6421 tctggatcta tctattgttc ttttcctcat cactttcccc tttttcgctc ttcttcttgt
     6481 cttttatttc tttctttttt ttaattgttc cctcgattgg ctatctacca aagaatccaa
     6541 acttaataca cgtatttatt tgtccaatta ccatggataa gaaatactca ataggcttag
     6601 atatcggcac aaatagcgtc ggatgggcgg tgatcactga tgaatataag gttccgtcta
     6661 aaaagttcaa ggttctggga aatacagacc gccacagtat caaaaaaaat cttatagggg
     6721 ctcttttatt tgacagtgga gagacagcgg aagcgactcg actcaaacgg acagctcgta
     6781 gaaggtatac acgtcggaag aatcgtattt gttatctaca ggagattttt tcaaatgaga
     6841 tggcgaaagt agatgatagt ttctttcatc gacttgaaga gtcttttttg gtggaagaag
     6901 acaagaagca tgaacgtcat cctatttttg gaaatatagt agatgaagtt gcttatcatg
     6961 agaaatatcc aactatctat catctgcgaa aaaaattggt agattctact gataaagcgg
     7021 atttgcgctt aatctatttg gccttagcgc atatgattaa gtttcgtggt cattttttga
     7081 ttgagggaga tttaaatcct gataatagtg atgtggacaa actatttatc cagttggtac
     7141 aaacctacaa tcaattattt gaagaaaacc ctattaacgc aagtggagta gatgctaaag
     7201 cgattctttc tgcacgattg agtaaatcaa gacgattaga aaatctcatt gctcagctcc
     7261 ccggtgagaa gaaaaatggc ttatttggga atctcattgc tttgtcattg ggtttgaccc
     7321 ctaattttaa atcaaatttt gatttggcag aagatgctaa attacagctt tcaaaagata
     7381 cttacgatga tgatttagat aatttattgg cgcaaattgg agatcaatat gctgatttgt
     7441 ttttggcagc taagaattta tcagatgcta ttttactttc agatatccta agagtaaata
     7501 ctgaaataac taaggctccc ctatcagctt caatgattaa acgctacgat gaacatcatc
     7561 aagacttgac tcttttaaaa gctttagttc gacaacaact tccagaaaag tataaagaaa
     7621 tcttttttga tcaatcaaaa aacggatatg caggttatat tgatggggga gctagccaag
     7681 aagaatttta taaatttatc aaaccaattt tagaaaaaat ggatggtact gaggaattat
     7741 tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac aacggctcta
     7801 ttccccatca aattcacttg ggtgagctgc atgctatttt gagaagacaa gaagactttt
     7861 atccattttt aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt
     7921 attatgttgg tccattggcg cgtggcaata gtcgttttgc atggatgact cggaagtctg
     7981 aagaaacaat taccccatgg aattttgaag aagttgtcga taaaggtgct tcagctcaat
     8041 catttattga acgcatgaca aactttgata aaaatcttcc aaatgaaaaa gtactaccaa
     8101 aacatagttt gctttatgag tattttacgg tttataacga attgacaaag gtcaaatatg
     8161 ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa gccattgttg
     8221 atttactctt caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa gattatttca
     8281 aaaaaataga atgttttgat agtgttgaaa tttcaggagt tgaagataga tttaatgctt
     8341 cattaggtac ctaccatgat ttgctaaaaa ttattaaaga taaagatttt ttggataatg
     8401 aagaaaatga agatatctta gaggatattg ttttaacatt gaccttattt gaagataggg
     8461 agatgattga ggaaagactt aaaacatatg ctcacctctt tgatgataag gtgatgaaac
     8521 agcttaaacg tcgccgttat actggttggg gacgtttgtc tcgaaaattg attaatggta
     8581 ttagggataa gcaatctggc aaaacaatat tagatttttt gaaatcagat ggttttgcca
     8641 atcgcaattt tatgcagctg atccatgatg atagtttgac atttaaagaa gacattcaaa
     8701 aagcacaagt gtctggacaa ggcgatagtt tacatgaaca tattgcaaat ttagctggta
     8761 gccctgctat taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag
     8821 taatggggcg gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa
     8881 ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag
     8941 aattaggaag tcagattctt aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa
     9001 agctctatct ctattatctc caaaatggaa gagacatgta tgtggaccaa gaattagata
     9061 ttaatcgttt aagtgattat gatgtcgatc acattgttcc acaaagtttc cttaaagacg
     9121 attcaataga caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg
     9181 ttccaagtga agaagtagtc aaaaagatga aaaactattg gagacaactt ctaaacgcca
     9241 agttaatcac tcaacgtaag tttgataatt taacgaaagc tgaacgtgga ggtttgagtg
     9301 aacttgataa agctggtttt atcaaacgcc aattggttga aactcgccaa atcactaagc
     9361 atgtggcaca aattttggat agtcgcatga atactaaata cgatgaaaat gataaactta
     9421 ttcgagaggt taaagtgatt accttaaaat ctaaattagt ttctgacttc cgaaaagatt
     9481 tccaattcta taaagtacgt gagattaaca attaccatca tgcccatgat gcgtatctaa
     9541 atgccgtcgt tggaactgct ttgattaaga aatatccaaa acttgaatcg gagtttgtct
     9601 atggtgatta taaagtttat gatgttcgta aaatgattgc taagtctgag caagaaatag
     9661 gcaaagcaac cgcaaaatat ttcttttact ctaatatcat gaacttcttc aaaacagaaa
     9721 ttacacttgc aaatggagag attcgcaaac gccctctaat cgaaactaat ggggaaactg
     9781 gagaaattgt ctgggataaa gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc
     9841 cccaagtcaa tattgtcaag aaaacagaag tacagacagg cggattctcc aaggagtcaa
     9901 ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa
     9961 aatatggtgg ttttgatagt ccaacggtag cttattcagt cctagtggtt gctaaggtgg
    10021 aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt actagggatc acaattatgg
    10081 aaagaagttc ctttgaaaaa aatccgattg actttttaga agctaaagga tataaggaag
    10141 ttaaaaaaga cttaatcatt aaactaccta aatatagtct ttttgagtta gaaaacggtc
    10201 gtaaacggat gctggctagt gccggagaat tacaaaaagg aaatgagctg gctctgccaa
    10261 gcaaatatgt gaatttttta tatttagcta gtcattatga aaagttgaag ggtagtccag
    10321 aagataacga acaaaaacaa ttgtttgtgg agcagcataa gcattattta gatgagatta
    10381 ttgagcaaat cagtgaattt tctaagcgtg ttattttagc agatgccaat ttagataaag
    10441 ttcttagtgc atataacaaa catagagaca aaccaatacg tgaacaagca gaaaatatta
    10501 ttcatttatt tacgttgacg aatcttggag ctcccgctgc ttttaaatat tttgatacaa
    10561 caattgatcg taaacgatat acgtctacaa aagaagtttt agatgccact cttatccatc
    10621 aatccatcac tggtctttat gaaacacgca ttgatttgag tcagctagga ggtgacggtg
    10681 gagggccaaa aaagaaaaga aaagttgaag atgcttctgc tcatcatcac catcaccatc
    10741 accattaaat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
    10801 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
    10861 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
    10921 gttttgggac gctcgaagtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
    10981 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
    11041 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
    11101 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
    11161 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
    11221 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
    11281 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
    11341 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
    11401 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
    11461 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
    11521 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    11581 caagaagatc ctttgatctt ttctac
//
