LOCUS       sequence-321816-       12385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 and contains natR854 guide which targets the natR 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12385)
  AUTHORS   Parts L, Batte A, Lopes M, Yuen MW, Laver M, San Luis BJ, Yue JX, 
            Pons C, Eray E, Aloy P, Liti G, van Leeuwen J
  TITLE     Natural variants suppress mutations in hundreds of essential genes.
  JOURNAL   Mol Syst Biol. 2021 May;17(5):e10138. doi: 10.15252/msb.202010138.
  PUBMED    34042294
REFERENCE   2  (bases 1 to 12385)
  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: "Mol Syst 
            Biol."; date: "2021-05"; volume: "17(5):e10138. doi"; pages: " 
            10.15252/msb.202010138"
COMMENT     Sequence Label: pML107-natR854
FEATURES             Location/Qualifiers
     source          1..12385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..269
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        290..364
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      369..388
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     terminator      complement(506..693)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     614..638
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(721..741)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3136..4854)
                     /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"
     regulatory      4851..4860
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4889..5555)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(4996..5019)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(5600..5616)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5601..5617)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5641..5660)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5642..5660)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5670..5687)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5670..5686)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5679..5701)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5827..6282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5914..5933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6124..6145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6583..6990
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6991..8085
                     /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(8579..8598)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8698..8720
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8758..8776)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8817..10163
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        10190..10294
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10295..10363,10364..11155)
                     /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(10513..10532)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11326..11914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11815..11834
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12068..12085
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12202..12223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12238..12243,12244..12261,12262..12268)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12276..12292
                     /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     12281..12303
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12300..12316
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12300..12316
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12335..12355
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12337..12355
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tctttgaaaa gataatgtat gattatgctt tcactcatat ttatacagaa acttgatgtt
       61 ttctttcgag tatatacaag gtgattacat gtacgtttga agtacaactc tagattttgt
      121 agtgccctct tgggctagcg gtaaaggtgc gcattttttc acaccctaca atgttctgtt
      181 caaaagattt tggtcaaacg ctgtagaagt gaaagttggt gcgcatgttt cggcgttcga
      241 aacttctccg cagtgaaaga taaatgatcc ggatggggtt caccctctgg ttttagagct
      301 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
      361 ggtggtgctt tttttgtttt ttatgtcttc gagtcatgta attagttata agcatgtgag
      421 ctaagacact gtaattgcca atctaaacga taccacggcc gctctagaga aatggggagc
      481 gatttgcagg catttgctcg gcatgccggt agaggtgtgg tcaataagag cgacctcatg
      541 ctatacctga gaaagcaacc tgacctacag gaaagagtta ctcaagaata agaattttcg
      601 ttttaaaacc taagagtcac tttaaaattt gtatacactt atttttttta taacttattt
      661 aataataaaa atcataaatc ataagaaatt cgcctcgaga gtctaggatc cggaactacc
      721 taccttgcgc tttttcttgg gagctccctc atcccctccg agctgtgaga ggtcaatccg
      781 agtttcgtaa agtccggtga tgctctggtg gatgagggtg gcgtccagca cttctttggt
      841 ggaggtatag cgtttccggt cgatggtggt gtcgaagtac ttgaatgcgg caggggcacc
      901 aagattggtg agggtgaaca ggtggatgat attctcagcc tgctcccgaa ttggtttgtc
      961 tctgtgctta ttgtatgcgc taaggacttt atcgaggttg gcatcagcga gaatgacgcg
     1021 tttagaaaac tcggaaatct gctcaatgat ctcgtccaga taatgcttat gctgctccac
     1081 aaacagttgc ttttgctcgt tgtcctcggg gctccccttc agcttctcgt aatggcttgc
     1141 aaggtagagg aaattcacgt acttggaggg gagagccagc tcattaccct tctgaagttc
     1201 gccagcggag gcgagcatcc gcttccgccc attctccagt tcgaacaggc tatactttgg
     1261 gagcttgatg ataaggtcct tcttgacttc cttgtaacct ttggcttcga gaaagtcaat
     1321 tgggttcttc tcgaaggagc tgcgctccat aatggtgata cccagcagtt ccttcacgct
     1381 cttgagcttc ttagactttc ctttttccac cttagccacg accaggacag aataagccac
     1441 agttggtgaa tcgaatccgc cgtacttctt agggtcccaa tccttcttcc ttgcaatgag
     1501 cttgtcggag ttgcgcttgg ggaggattga ttccttagaa aagcctccgg tttgcacctc
     1561 ggttttcttc acgatgttca cttgaggcat agaaagcact ttgcgcacgg tagcgaaatc
     1621 tctgcccttg tcccacacga tttcgccagt ctccccatta gtctcgatga gcggcctctt
     1681 ccggatctca ccgtttgcaa gggtgatttc ggtcttgaag aaattcatga tgttgctgta
     1741 gaagaagtac tttgcggtgg cctttccgat ctcctgctca gatttggcaa tcatcttgcg
     1801 cacgtcgtag accttgtagt ctccgtacac gaactcgctt tccagctttg ggtacttctt
     1861 gattagggcg gttcccacca cggcattcag atatgcgtca tgagcatgat gatagttatt
     1921 gatctcccgc actttgtaga attgaaagtc ctttctgaaa tcgctgacca gcttggactt
     1981 cagggtaatc accttcacct cgcggataag tttgtcgttt tcgtcgtact tggtattcat
     2041 gcgtgagtca aggatttggg ctacatgttt agtaatctgc cgagtctcca caagttgccg
     2101 tttaatgaat cctgctttgt ccagttcgct cagtccgccc ctctcagcct tagtgagatt
     2161 gtcgaacttc cgctgagtga ttagctttgc gttcagaagc tggcgccaat agttcttcat
     2221 ctttttcacg acttcctctg agggcacgtt atctgacttt ccccggttct tgtctgagcg
     2281 agtcagcacc ttattgtcaa ttgagtcgtc tttgaggaag ctctgaggga cgatgtggtc
     2341 cacgtcgtag tctgagagcc tgttgatatc aagctcttgg tccacataca tatcgcgtcc
     2401 attttgaagg tagtacagat agagcttctc attctgaagc tgggtgtttt ccaccgggtg
     2461 ctctttaagg atctgtgacc cgagctcctt aatcccttcc tcaattcttt tcatccgctc
     2521 cctgctgttc ttctgtccct tctgggtagt ctggttttcc ctggccatct cgatgacaat
     2581 gttctctggt ttatggcgac ccatcacctt gaccagctcg tccacgactt tcacggtctg
     2641 gagaataccc ttcttaatag cgggtgaacc agccagattt gcgatatgct cgtgcagtga
     2701 gtcaccttgt ccagacactt gggccttctg gatgtcctcc ttaaaggtca gggaatcatc
     2761 atggataagt tgcatgaagt tgcgattagc gaatccgtcg ctcttaagga aatcaagaat
     2821 ggtctttccg ctctgtttgt cccggattcc gttgatgagt ttgcggctaa ggcgtcccca
     2881 tccggtatat cttctccgtt taagttgctt catcacttta tcgtcgaaca gatgggcgta
     2941 ggtcttaagc ctctcctcga tcatttcccg gtcctcgaac agagtgagag tcaggacaat
     3001 gtcctcaagg atgtcctctt tctcctcatt gtccagaaaa tccttgtcct tgatgatctt
     3061 aagcagatca tggtaggtac ccaggcttgc gttgaaccga tcttcgaccc cgctaatctc
     3121 cacagagtcg aagcattcaa ttttcttgaa atagtcctcc ttcagctgct tcacggtcac
     3181 ctttctattg gtcttgaaca gaaggtccac aattgccttc ttttgttcgc ctgacaggaa
     3241 tgcgggcttc ctcatccctt cggtgacata tttcaccttg gtcagctcgt tgtacacggt
     3301 aaagtactcg tagagcaggg aatgtttcgg caggaccttc tcattgggga ggttcttatc
     3361 gaagttagtc atccgttcga tgaatgactg agcgctggct cccttgtcca ccacctcttc
     3421 gaagttccag ggagtgatag tttcttctga ctttctggtc atccaagcaa accggctatt
     3481 tcctctggcg aggggtccca cgtagtaggg gatgcggaaa gtgagaattt tctcaatctt
     3541 ctccctgttg tccttaagaa aggggtagaa gtcctcttgg cgccgaagga tggcgtgaag
     3601 ctccccaaga tggatctggt ggggaatgct cccattgtcg aaagtgcgct gcttccgcag
     3661 cagatcctcc cgattcagtt tcacgaggag ctcttcggtg ccgtccatct tctcaaggat
     3721 aggtttgata aacttataaa attcctcttg ggaggcacct ccgtcgatgt agccggcgta
     3781 tccattcttt gactggtcaa agaagatttc cttgtacttc tccggaagct gctgccggac
     3841 gagtgcctta agcagggtca ggtcctggtg gtgctcatca taccgcttaa tcattgaagc
     3901 tgaaagaggg gccttggtga tttcagtgtt cacgcggaga atgtcagaca gcaggatggc
     3961 gtcagaaaga ttcttagcag caaggaaaag gtcggcgtat tggtcaccga tctgggccag
     4021 gaggttgtcg agatcgtcat cataggtgtc cttggacagt tgcagcttgg catcctctgc
     4081 caggtcgaaa ttagacttga aattgggagt cagcccaagt gaaagagcaa tcaggttccc
     4141 gaacagtcca tttttctttt cacccggcag ctgagcgata agattctcaa gtctgcggct
     4201 ctttgacagc cgggctgaca gaatggcctt ggcatcgact ccgcttgcat tgatggggtt
     4261 ctcttcaaag agctggttgt aagtctgcac cagctggatg aacagtttat ccacatcgct
     4321 attgtcagga ttgaggtcgc cctcgatcag gaagtggcct ctgaacttaa tcatgtgggc
     4381 cagggccagg tagataagcc ggaggtctgc cttgtcggtt gagtccacca gctttttgcg
     4441 aagatggtag atggttgggt atttctcgtg ataggccacc tcgtcgacga tattaccgaa
     4501 gatgggatgc cgttcatgct tcttatcctc ttccacgagg aatgattctt ccagcctatg
     4561 gaagaagctg tcgtcgacct ttgccatttc gttggagaat atctcttgaa ggtaacaaat
     4621 ccggttcttc ctcctggtgt accgccgtct agcagtccgc ttgaggcggg ttgcttcagc
     4681 ggtctcccct gaatcaaaga ggagagctcc gatcagattt ttcttgatgg aatgccggtc
     4741 ggtgttcccg agcaccttga acttcttaga gggcaccttg tactcgtcag tgatgacggc
     4801 ccacccgacg ctattagtcc cgatgtccag tccgatagaa tacttcttgt ccatggtggc
     4861 gagatctaat tcgatccata tcagatcttt tgtttgttta tgtgtgttta ttcgaaacta
     4921 agttcttggt gttttaaaac taaaaaaaag actaactata aaagtagaat ttaagaagtt
     4981 taagaaatag atttacagaa ttacaatcaa tacctaccgt ctttatatac ttattagtca
     5041 agtaggggaa taatttcagg gaactggttt caaccttttt tttcagcttt ttccaaatca
     5101 gagagagcag aaggtaatag aaggtgtaag aaaatgagat agatacatgc gtgggtcaat
     5161 tgccttgtgt catcatttac tccaggcagg ttgcatcact ccattgaggt tgtgcccgtt
     5221 ttttgcctgt ttgtgcccct gttctctgta gttgcgctaa gagaatggac ctatgaactg
     5281 atggttggtg aagaaaacaa tattttggtg ctgggattct ttttttttct ggatgccagc
     5341 ttaaaaagcg ggctccatta tatttagtgg atgccaggaa taaactgttc acccagacac
     5401 ctacgatgtt atatattctg tgtaacccgc cccctatttt gggcatgtac gggttacagc
     5461 agaattaaaa ggctaatttt ttgactaaat aaagttagga aaatcactac tattaattat
     5521 ttacgtattc tttgaaatgg cagtattgat aatgataaac tcgaactgac tagtagactg
     5581 aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt acccaattcg
     5641 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     5701 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5761 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5821 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5881 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5941 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     6001 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     6061 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     6121 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     6181 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6241 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     6301 ccttacgcat ctgtgcggta tttcacaccg catatcgacc ggtcgaggag aacttctagt
     6361 atatctacat acctaatatt attgccttat taaaaatgga atcccaacaa ttacatcaaa
     6421 atccacattc tcttcaaaat caattgtcct gtacttcctt gttcatgtgt gttcaaaaac
     6481 gttatattta taggataatt atactctatt tctcaacaag taattggttg tttggccgag
     6541 cggtctaagg cgcctgattc aagaaatatc ttgaccgcag ttaactgtgg gaatactcag
     6601 gtatcgtaag atgcaagagt tcgaatctct tagcaaccat tatttttttc ctcaacataa
     6661 cgagaacaca caggggcgct atcgcacaga atcaaattcg atgactggaa attttttgtt
     6721 aatttcagag gtcgcctgac gcatatacct ttttcaactg aaaaattggg agaaaaagga
     6781 aaggtgagag cgccggaacc ggcttttcat atagaataga gaagcgttca tgactaaatg
     6841 cttgcatcac aatacttgaa gttgacaata ttatttaagg acctattgtt ttttccaata
     6901 ggtggttagc aatcgtctta ctttctaact tttcttacct tttacatttc agcaatatat
     6961 atatatatat ttcaaggata taccattcta atgtctgccc ctaagaagat cgtcgttttg
     7021 ccaggtgacc acgttggtca agaaatcaca gccgaagcca ttaaggttct taaagctatt
     7081 tctgatgttc gttccaatgt caagttcgat ttcgaaaatc atttaattgg tggtgctgct
     7141 atagatgcta caggtgttcc acttccagat gaggcgctgg aagcctccaa gaaggctgat
     7201 gccgttttgt taggtgctgt gggtggtcct aaatggggta caggtagtgt tagacctgaa
     7261 caaggtttac taaaaatccg taaagaactt caattgtacg ccaacttaag accatgtaac
     7321 tttgcatccg actctctttt agacttatct ccaatcaagc cacaatttgc taaaggtact
     7381 gacttcgttg ttgtcagaga attagtggga ggtatttact ttggtaagag aaaggaagac
     7441 gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt gcaaagaatc
     7501 acaagaatgg ccgctttcat ggccctacaa catgagccac cattgcctat ttggtccttg
     7561 gataaagcta atgttttggc ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc
     7621 aagaacgaat ttcctacatt gaaggttcaa catcaattga ttgattctgc cgccatgatc
     7681 ctagttaaga acccaaccca cctaaatggt attataatca ccagcaacat gtttggtgat
     7741 atcatctccg atgaagcctc cgttatccca ggttccttgg gtttgttgcc atctgcgtcc
     7801 ttggcctctt tgccagacaa gaacaccgca tttggtttgt acgaaccatg ccacggttct
     7861 gctccagatt tgccaaagaa taaggtcaac cctatcgcca ctatcttgtc tgctgcaatg
     7921 atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg ccattgaaga tgcagttaaa
     7981 aaggttttgg atgcaggtat cagaactggt gatttaggtg gttccaacag taccaccgaa
     8041 gtcggtgatg ctgtcgccga agaagttaag aaaatccttg cttaaaaaga ttctcttttt
     8101 ttatgatatt tgtacataaa ctttataaat gaaattcata atagaaacga cacgaaatta
     8161 caaaatggaa tatgttcata gggtagacga aactatatac gcaatctaca tacatttatc
     8221 aagaaggaga aaaaggagga tgtaaaggaa tacaggtaag caaattgata ctaatggctc
     8281 aacgtgataa ggaaaaagaa ttgcacttta acattaatat tgacaaggag gagggcacca
     8341 cacaaaaagt taggtgtaac agaaaatcat gaaactatga ttcctaattt atatattgga
     8401 ggattttctc taaaaaaaaa aaaatacaac aaataaaaaa cactcaatga cctgaccatt
     8461 tgatggagtt taagtcaata ccttcttgaa ccatttccca taatggtgaa agttccctca
     8521 agaattttac tctgtcagaa acggccttaa cgacgtagtc gatatggtgc actctcagta
     8581 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     8641 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     8701 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     8761 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagtatgatc
     8821 caatatcaaa ggaaatgata gcattgaagg atgagactaa tccaattgag gagtggcagc
     8881 atatagaaca gctaaagggt agtgctgaag gaagcatacg ataccccgca tggaatggga
     8941 taatatcaca ggaggtacta gactaccttt catcctacat aaatagacgc atataagtac
     9001 gcatttaagc ataaacacgc actatgccgt tcttctcatg tatatatata tacaggcaac
     9061 acgcagatat aggtgcgacg tgaacagtga gctgtatgtg cgcagctcgc gttgcatttt
     9121 cggaagcgct cgttttcgga aacgctttga agttcctatt ccgaagttcc tattctctag
     9181 ctagaaagta taggaacttc agagcgcttt tgaaaaccaa aagcgctctg aagacgcact
     9241 ttcaaaaaac caaaaacgca ccggactgta acgagctact aaaatattgc gaataccgct
     9301 tccacaaaca ttgctcaaaa gtatctcttt gctatatatc tctgtgctat atccctatat
     9361 aacctaccca tccacctttc gctccttgaa cttgcatcta aactcgacct ctacattttt
     9421 tatgtttatc tctagtatta ctctttagac aaaaaaattg tagtaagaac tattcataga
     9481 gtgaatcgaa aacaatacga aaatgtaaac atttcctata cgtagtatat agagacaaaa
     9541 tagaagaaac cgttcataat tttctgacca atgaagaatc atcaacgcta tcactttctg
     9601 ttcacaaagt atgcgcaatc cacatcggta tagaatataa tcggggatgc ctttatcttg
     9661 aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc gggaagtgga gtcaggcttt
     9721 ttttatggaa gagaaaatag acaccaaagt agccttcttc taaccttaac ggacctacag
     9781 tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa aggagaaaaa aagtaatcta
     9841 agatgctttg ttagaaaaat agcgctctcg ggatgcattt ttgtagaaca aaaaagaagt
     9901 atagattctt tgttggtaaa atagcgctct cgcgttgcat ttctgttctg taaaaatgca
     9961 gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ttacaaaaat
    10021 gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt tgcatttctg ttctgtaaaa
    10081 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttctaca
    10141 aaatgaagca cagatgcttc gttcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
    10201 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
    10261 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
    10321 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
    10381 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
    10441 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
    10501 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
    10561 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
    10621 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
    10681 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
    10741 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
    10801 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
    10861 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
    10921 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
    10981 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
    11041 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
    11101 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
    11161 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
    11221 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
    11281 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
    11341 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
    11401 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
    11461 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
    11521 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
    11581 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
    11641 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
    11701 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
    11761 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
    11821 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
    11881 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
    11941 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
    12001 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
    12061 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
    12121 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
    12181 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
    12241 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
    12301 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
    12361 aaagctggag ctccaccgcg gacaa
//
