LOCUS       sequence-330960-       10916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression of SpCas9 in yeast after assembly with PID 
            fragments: TEF1p-SpCas9::KanR-1xNLS-CYC1t (KanR cassette in place of
            the PID).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10916)
  AUTHORS   Goldberg GW, Spencer JM, Giganti DO, Camellato BR, Agmon N, Ichikawa
            DM, Boeke JD, Noyes MB
  TITLE     Engineered dual selection for directed evolution of SpCas9 PAM 
            specificity.
  JOURNAL   Nat Commun. 2021 Jan 13;12(1):349. doi: 10.1038/s41467-020-20650-x.
  PUBMED    33441553
REFERENCE   2  (bases 1 to 10916)
  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: "Nat 
            Commun."; date: "2021-01-13"; volume: "12(1):349. doi"; pages: " 
            10.1038/s41467-020-20650-x"
COMMENT     Sequence Label: pGG431
FEATURES             Location/Qualifiers
     source          1..10916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     403..419
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     403..419
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             431..2149
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             3018..3029
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(3734..4528)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3845..3864)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4455..4474
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             4776..4796
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4812..5059
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4828..4846)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5077..5096)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5078..5096)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5106..5123)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5106..5122)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5115..5137)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5263..5718
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5350..5369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5560..5581
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6019..6426
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6427..7521
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(8015..8034)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8134..8152)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    8194..8697
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        8734..8838
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8839..8907,8908..9699)
                     /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     complement(9057..9076)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9870..10458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10359..10378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10612..10629
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10746..10767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10782..10787,10788..10805,10806..10812)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10820..10836
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     10825..10847
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10844..10860
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10844..10860
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10879..10899
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10881..10899
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 catagcttca aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc
       61 atcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct
      121 ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt
      181 ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa
      241 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
      301 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
      361 cttgctcatt agaaagaaag catagcaatc taatctaagt tttctagaac tagtggatcc
      421 cccgggaaaa atggacaaga agtactccat tgggctcgat atcggcacaa acagcgtcgg
      481 ttgggccgtc attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa
      541 taccgatcgc cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga
      601 gacggccgaa gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa
      661 tcggatctgc tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt
      721 cttccatagg ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc
      781 aatctttggc aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca
      841 tctgaggaag aagcttgtag acagcactga taaggctgac ttgcggttga tctatctcgc
      901 gctggcgcat atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga
      961 caacagcgat gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga
     1021 agagaacccg atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc
     1081 caaatcccgg cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct
     1141 gtttggtaat cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga
     1201 cctggccgaa gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa
     1261 tctgctggcc cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc
     1321 agacgccatt ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct
     1381 gagcgctagt atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc
     1441 ccttgtcaga cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa
     1501 tggctacgcc ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa
     1561 gcccatcttg gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga
     1621 tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg
     1681 cgaactgcac gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag
     1741 ggaaaagatt gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg
     1801 gggaaattcc agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa
     1861 cttcgaggaa gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa
     1921 ctttgataaa aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta
     1981 cttcacagtt tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc
     2041 agcattcctg tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg
     2101 gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc
     2161 tgttgaaatc agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct
     2221 cctgaaaatc attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga
     2281 ggacattgtc ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa
     2341 aacttacgct catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac
     2401 aggatggggg cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa
     2461 gacaatcctg gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat
     2521 ccatgatgac tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg
     2581 ggacagtctt cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat
     2641 actgcagacc gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga
     2701 gaatatcgtt atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag
     2761 tagggaaagg atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa
     2821 ggaacaccca gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca
     2881 gaacggcagg gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga
     2941 cgtggatcat atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt
     3001 gacaagatcc gataaaaata gagggaagag tgataacgtc ccctcagaag aagttgtcaa
     3061 gaaaatgaaa aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt
     3121 cgataatctg actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat
     3181 caaaaggcag cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc
     3241 acgcatgaac accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac
     3301 tctgaagtct aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga
     3361 gatcaacaat taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact
     3421 tatcaaaaaa tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga
     3481 tgttaggaaa atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt
     3541 cttttacagc aatattatga attttttcaa gaccgagatt acactggcca atggagagat
     3601 tcggaagcga ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg
     3661 tagggatttc gcgacagtcc ggaaggtcct gtccatgccg caagtcagct agcctcgaac
     3721 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     3781 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     3841 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     3901 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     3961 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     4021 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     4081 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     4141 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     4201 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     4261 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     4321 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     4381 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     4441 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     4501 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     4561 atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     4621 agggcttccc aaccttacca gagggcgccc cagctggcaa ttccgacgtc acccgtaggc
     4681 atcaaataaa acgaaaggct cagtcgaggg cccaatatgg gctctatgaa acaagaatcg
     4741 acctctctca gctcggtgga gacagcaggg ctgaccccaa gaagaagagg aaggtgtgat
     4801 ctcttctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     4861 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     4921 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     4981 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     5041 ctcgaaggct ttaatttgcg gccggtaccc aattcgccct atagtgagtc gtattacgcg
     5101 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5161 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5221 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     5281 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5341 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5401 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5461 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5521 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5581 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5641 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5701 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     5761 acaccgcata tcgaccggtc gaggagaact tctagtatat ctacatacct aatattattg
     5821 ccttattaaa aatggaatcc caacaattac atcaaaatcc acattctctt caaaatcaat
     5881 tgtcctgtac ttccttgttc atgtgtgttc aaaaacgtta tatttatagg ataattatac
     5941 tctatttctc aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga
     6001 aatatcttga ccgcagttaa ctgtgggaat actcaggtat cgtaagatgc aagagttcga
     6061 atctcttagc aaccattatt tttttcctca acataacgag aacacacagg ggcgctatcg
     6121 cacagaatca aattcgatga ctggaaattt tttgttaatt tcagaggtcg cctgacgcat
     6181 ataccttttt caactgaaaa attgggagaa aaaggaaagg tgagagcgcc ggaaccggct
     6241 tttcatatag aatagagaag cgttcatgac taaatgcttg catcacaata cttgaagttg
     6301 acaatattat ttaaggacct attgtttttt ccaataggtg gttagcaatc gtcttacttt
     6361 ctaacttttc ttacctttta catttcagca atatatatat atatatttca aggatatacc
     6421 attctaatgt ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa
     6481 atcacagccg aagccattaa ggttcttaaa gctatttctg atgttcgttc caatgtcaag
     6541 ttcgatttcg aaaatcattt aattggtggt gctgctatcg atgctacagg tgttccactt
     6601 ccagatgagg cgctggaagc ctccaagaag gctgatgccg ttttgttagg tgctgtgggt
     6661 ggtcctaaat ggggtaccgg tagtgttaga cctgaacaag gtttactaaa aatccgtaaa
     6721 gaacttcaat tgtacgccaa cttaagacca tgtaactttg catccgactc tcttttagac
     6781 ttatctccaa tcaagccaca atttgctaaa ggtactgact tcgttgttgt cagagaatta
     6841 gtgggaggta tttactttgg taagagaaag gaagacgatg gtgatggtgt cgcttgggat
     6901 agtgaacaat acaccgttcc agaagtgcaa agaatcacaa gaatggccgc tttcatggcc
     6961 ctacaacatg agccaccatt gcctatttgg tccttggata aagctaatgt tttggcctct
     7021 tcaagattat ggagaaaaac tgtggaggaa accatcaaga acgaattccc tacattgaag
     7081 gttcaacatc aattgattga ttctgccgcc atgatcctag ttaagaaccc aacccaccta
     7141 aatggtatta taatcaccag caacatgttt ggtgatatca tctccgatga agcctccgtt
     7201 atcccaggtt ccttgggttt gttgccatct gcgtccttgg cctctttgcc agacaagaac
     7261 accgcatttg gtttgtacga accatgccac ggttctgctc cagatttgcc aaagaataag
     7321 gtcaacccta tcgccactat cttgtctgct gcaatgatgt tgaaattgtc attgaacttg
     7381 cctgaagaag gtaaggccat tgaagatgca gttaaaaagg ttttggatgc aggtatcaga
     7441 actggtgatt taggtggttc caacagtacc accgaagtcg gtgatgctgt cgccgaagaa
     7501 gttaagaaaa tccttgctta aaaagattct ctttttttat gatatttgta cataaacttt
     7561 ataaatgaaa ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt
     7621 agacgaaact atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta
     7681 aaggaataca ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc
     7741 actttaacat taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa
     7801 aatcatgaaa ctatgattcc taatttatat attggaggat tttctctaaa aaaaaaaaaa
     7861 tacaacaaat aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt
     7921 cttgaaccat ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg
     7981 ccttaacgac gtagtcgata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     8041 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     8101 cggcatccgc ttacagacaa gctgtgacaa agggcctcgt gatacgccta tttttatagg
     8161 ttaatgtcat gataataatg gtttcttagg acggatcgct tgcctgtaac ttacacgcgc
     8221 ctcgtatctt ttaatgatgg aataatttgg gaatttactc tgtgtttatt tatttttatg
     8281 ttttgtattt ggattttaga aagtaaataa agaaggtaga agagttacgg aatgaagaaa
     8341 aaaaaataaa caaaggttta aaaaatttca acaaaaagcg tactttacat atatatttat
     8401 tagacaagaa aagcagatta aatagatata cattcgatta acgataagta aaatgtaaaa
     8461 tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc ggcattaata
     8521 cctgagagca ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc ctagagtctt
     8581 ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt
     8641 taatttatat atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa
     8701 aaggacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     8761 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     8821 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     8881 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     8941 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     9001 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     9061 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     9121 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     9181 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     9241 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     9301 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     9361 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     9421 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     9481 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     9541 gccggtgagc gtggatctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     9601 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     9661 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     9721 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     9781 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     9841 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     9901 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     9961 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
    10021 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
    10081 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
    10141 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
    10201 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
    10261 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
    10321 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
    10381 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
    10441 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
    10501 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
    10561 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
    10621 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
    10681 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
    10741 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
    10801 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
    10861 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagct
//
