LOCUS Exported 9983 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Cas9 fused with GFP and a sgRNA targeting the Cryptosporidium parvum UPRT gene. ACCESSION . VERSION . KEYWORDS pACT1:Cas9-GFP, U6:sgUPRT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9983) AUTHORS Wilke G, Funkhouser-Jones LJ, Wang Y, Ravindran S, Wang Q, Beatty WL, Baldridge MT, VanDussen KL, Shen B, Kuhlenschmidt MS, Kuhlenschmidt TB, Witola WH, Stappenbeck TS, Sibley LD TITLE A Stem-Cell-Derived Platform Enables Complete Cryptosporidium Development In Vitro and Genetic Tractability. JOURNAL Cell Host Microbe. 2019 Jun 18. pii: S1931-3128(19)30252-5. doi: 10.1016/j.chom.2019.05.007. PUBMED 31231046 REFERENCE 2 (bases 1 to 9983) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9983 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(10..28) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 96..200 /gene="bla" /label=AmpR promoter CDS 201..1061 /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(419..438) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1232..1820 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1721..1740 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1974..1991 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2108..2129 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2144..2174 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2182..2198 /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 2187..2209 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2206..2222 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2206..2222 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS complement(2814..3530) /codon_start=1 /product="enhanced GFP (EGFP) derivative that exhibits robust folding and enhanced photostability (Zhong et al., 2019)" /label=SiriusGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(2859..2880) /label=EGFP-C /note="EGFP, forward primer" primer_bind 3467..3488 /label=EGFP-N /note="EGFP, reverse primer" CDS complement(3570..3590) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(3597..3617) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(3636..3662) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS complement(3666..7769) /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" misc_RNA 9437..9512 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter complement(9552..9570) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(9553..9570) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(9589..9606) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(9589..9605) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9598..9620) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(9814..9833) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 9933..9955 /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct 961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac 1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt 1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc 2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca 2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc 2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg 2221 accatgatta cgccaagctt gcatgcctgc atagtatatc cgcctgtcac gcggatgacc 2281 cgggttcaat tcccggactt ggagcttttt ttaaaaaatt tgttttgcct aaatttggac 2341 cgctcttttt tattgttaaa tggcagctct attttcggac ttgaactttg gtaataaaca 2401 tcacctgaca aatcaactta attaatttct aatactgtgc tcaggaaaac aacacgcaca 2461 taatttaaac actttcgttt aaaaagaagg aaaaaatcgt atcgaactac acattactta 2521 gtaactattt tgattcaaac tagcccacct tctatccata atattagtta tcattgacta 2581 acaattataa tagaattttc ttgctaaatt tagagtttat tagcattgat tagacaattt 2641 atcatctttt tttttagctt tcaaatgaga atagaactga gatctgaata atgtaataag 2701 aaattgtcta caagaagtga taacttgtgt aattcatatt tctatttaag ttgcaatacg 2761 tatctgtgtt caggaaataa gctgttaaga acaactccaa gaacaatatt gatctacttg 2821 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg 2881 tgatcgcgct tctcgttggg gtctttgctc agcttggact gggtgctcag gtagtggttg 2941 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc 3001 tgcacgctgc cgtcctcgac gttgtggcgg atcttgaagt tcgccttgat gccgttcttc 3061 tgcttgtcgg ccgtgatata gacgttgtgg ctgttgaagt tgtactccag cttgtgcccc 3121 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg 3181 tcgccctcga acttcacctc ggcgcgggtc ttgtaggtgc cgtcgtcctt gaaggagatg 3241 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg 3301 tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc 3361 cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgttggtg 3421 gcatcgccct cgccctcgcc gcgcacgctg aacttgtggc cgtttacgtc gccgtccagc 3481 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac ggatccgctg 3541 ctgccgttgc tcccactacc aatgccgtcc actttccgct ttttcttagg atcttccacc 3601 ttgcgttttt tcttggggga gccgctgccc aggctggcat aatcgggcac gtcataggga 3661 taggcgtcgc ctcccagctg agacaggtcg atccgtgtct cgtacaggcc ggtgatgctc 3721 tggtggatca gggtggcgtc cagcacctct ttggtgctgg tgtacctctt ccggtcgatg 3781 gtggtgtcaa agtacttgaa ggcggcaggg gctcccagat tggtcagggt aaacaggtgg 3841 atgatattct cggcctgctc tctgataggc ttgtctctgt gcttgttgta ggcgctcagc 3901 accttgtcca gattagcgtc ggccaggatc actctcttgg agaactcgct gatctgctcg 3961 atgatctcgt ccaggtagtg tttgtgctgt tccacaaaca gctgtttctg ctcattatcc 4021 tcgggggagc ccttcagctt ctcatagtgg ctggccaggt acaggaagtt cacatatttg 4081 gagggcaggg ccagttcgtt tcccttctgc agttcgccgg cagaggccag cattctcttc 4141 cggccgtttt ccagctcgaa cagggagtac ttaggcagct tgatgatcag gtcctttttc 4201 acttctttgt agcccttggc ttccagaaag tcgatgggat tcttctcgaa gctgcttctt 4261 tccatgatgg tgatccccag cagctctttc acactcttca gtttcttgga cttgcccttt 4321 tccactttgg ccaccaccag cacagaatag gccacggtgg ggctgtcgaa gccgccgtac 4381 ttcttagggt cccagtcctt ctttctggcg atcagcttgt cgctgttcct cttgggcagg 4441 atagactctt tgctgaagcc gcctgtctgc acctcggtct ttttcacgat attcacttgg 4501 ggcatagaca gcactttccg cacggtggca aagtcccggc ccttatccca cacgatctcg 4561 cctgtttcgc cgtttgtctc gatcagaggc cgcttccgga tctcgccgtt ggccagggta 4621 atctcggtct tgaaaaagtt catgatgttg ctgtagaaga agtacttggc ggtagccttg 4681 ccgatttcct gctcgctctt ggcgatcatc ttccgcacgt cgtacacctt gtagtcgccg 4741 tacacgaact cgctttccag cttagggtac tttttgatca gggcggttcc cacgacggcg 4801 ttcaggtagg cgtcgtgggc gtggtggtag ttgttgatct cgcgcacttt gtaaaactgg 4861 aaatccttcc ggaaatcgga caccagcttg gacttcaggg tgatcacttt cacttcccgg 4921 atcagtttgt cgttctcgtc gtacttagtg ttcatccggg agtccaggat ctgtgccacg 4981 tgctttgtga tctgccgggt ttccaccagc tgtctcttga tgaagccggc cttatccagt 5041 tcgctcaggc cgcctctctc ggccttggtc agattgtcga acttcctctg ggtaatcagc 5101 ttggcattca gcagctggcg ccagtagttc ttcatcttct tcacgacctc ttcggagggc 5161 acgttgtcgc tcttgccccg gttcttgtcg ctccgagtca gcactttgtt atcgatggag 5221 tcgtccttca gaaagctctg aggcacgata tggtccacat cgtagtcgga cagccggttg 5281 atgtccagtt cctggtccac gtacatatcc cgcccattct gcaggtagta caggtacagc 5341 ttctcgttct gcagctgggt gttttccacg gggtgttctt tcaggatctg gctgcccagc 5401 tctttgatgc cctcttcgat ccgcttcatt ctctcgcggc tgttcttctg tcccttctgg 5461 gtggtctggt tctctctggc catttcgatc acgatgttct cgggcttgtg ccggcccatc 5521 actttcacga gctcgtccac caccttcact gtctgcagga tgcccttctt aatggcgggg 5581 ctgccggcca gattggcaat gtgctcgtgc aggctatcgc cctggccgga cacctgggct 5641 ttctggatgt cctctttaaa ggtcaggctg tcgtcgtgga tcagctgcat gaagtttctg 5701 ttggcgaagc cgtcggactt caggaaatcc aggattgtct tgccggactg cttgtcccgg 5761 atgccgttga tcagcttccg gctcagcctg ccccagccgg tgtatctccg ccgcttcagc 5821 tgcttcatca ctttgtcgtc gaacaggtgg gcataggttt tcagccgttc ctcgatcatc 5881 tctctgtcct caaacagtgt cagggtcagc acgatatctt ccagaatgtc ctcgttttcc 5941 tcattgtcca ggaagtcctt gtccttgata attttcagca gatcgtggta tgtgcccagg 6001 gaggcgttga accgatcttc cacgccggag atttccacgg agtcgaagca ctcgattttc 6061 ttgaagtagt cctctttcag ctgcttcacg gtcactttcc ggttggtctt gaacagcagg 6121 tccacgatgg cttttttctg ctcgccgctc aggaaggcgg gctttctcat tccctcggtc 6181 acgtatttca ctttggtcag ctcgttgtac acggtgaagt actcgtacag caggctgtgc 6241 ttgggcagca ccttctcgtt gggcaggttc ttatcgaagt tggtcatccg ctcgatgaag 6301 ctctgggcgc tggcgccctt gtccaccact tcctcgaagt tccagggggt gatggtttcc 6361 tcgctctttc tggtcatcca ggcgaatctg ctgtttcccc tggccagagg gcccacgtag 6421 taggggatgc ggaaggtcag gatcttctcg atcttttccc ggttgtcctt caggaatggg 6481 taaaaatctt cctgccgccg cagaatggcg tgcagctctc ccaggtggat ctggtggggg 6541 atgctgccgt tgtcgaaggt ccgctgcttc cgcagcaggt cctctctgtt cagcttcacg 6601 agcagttcct cggtgccgtc catcttttcc aggatgggct tgatgaactt gtagaactct 6661 tcctggctgg ctccgccatc gatgtagccg gcgtagccgt tcttgctctg gtcgaagaaa 6721 atctctttgt acttctcagg cagctgctgc cgcacgagag ctttcagcag ggtcaggtcc 6781 tggtggtgct cgtcgtatct cttgatcata gaggcgctca ggggggcctt ggtgatctcg 6841 gtgttcactc tcaggatgtc gctcagcagg atggcgtcgg acaggttctt ggcggccaga 6901 aacaggtcgg cgtactggtc gccgatctgg gccagcaggt tgtccaggtc gtcgtcgtag 6961 gtgtccttgc tcagctgcag tttggcatcc tcggccaggt cgaagttgct cttgaagttg 7021 ggggtcaggc ccaggctcag ggcaatcagg ttgccgaaca ggccattctt cttctcgccg 7081 ggcagctggg cgatcagatt ttccagccgt ctgctcttgc tcagtctggc agacaggatg 7141 gccttggcgt ccacgccgct ggcgttgatg gggttttcct cgaacagctg gttgtaggtc 7201 tgcaccagct ggatgaacag cttgtccacg tcgctgttgt cggggttcag gtcgccctcg 7261 atcaggaagt ggccccggaa cttgatcatg tgggccaggg ccagatagat cagccgcagg 7321 tcggccttgt cggtgctgtc caccagtttc tttctcaggt ggtagatggt ggggtacttc 7381 tcgtggtagg ccacctcgtc cacgatgttg ccgaagatgg ggtgccgctc gtgcttctta 7441 tcctcttcca ccaggaagga ctcttccagt ctgtggaaga agctgtcgtc caccttggcc 7501 atctcgttgc tgaagatctc ttgcagatag cagatccggt tcttccgtct ggtgtatctt 7561 cttctggcgg ttctcttcag ccgggtggcc tcggctgttt ctccgctgtc gaacagcagg 7621 gcgccgatca ggttcttctt gatgctgtgc cggtcggtgt tgcccagcac cttgaatttc 7681 ttgctgggca ccttgtactc gtcggtgatc acggcccagc ccacagagtt ggtgccgatg 7741 tccaggccga tgctgtactt cttgtccatc ttttgttact tgtgtaatat gcccgaaatc 7801 tatgaaaaaa aaaaatctat attgtaagta agctatatat aaaatataaa aactgctcta 7861 ttaaactact actttgcttg ccgtacaatt cttattatct ctttatccaa gtatttaaag 7921 atatgtatac gcacgcagga aaataaaaat atactttgga tttttaatgc gcactgtacg 7981 tatcgtaaaa agattcatct cactttttac atgttgtcta ttctaaattt atacttgatt 8041 tttatttaat ctctaaactt tgtcatgtaa tcgtctatta atacgtattg gcgcgcgtct 8101 atgcatggtg ggcggcaagg tgcatgcaat ttgacataaa tgcattcaat gcaagattta 8161 tatttaatat aaaatttggt cttttttttt tactaaaatc taaccttttt ttatttacat 8221 aactaagatt cgtttaaatt aatacgtttc acgtcgtaat atttatcaag taatgtatgt 8281 agcattgtat acatgtgatg tacgaatttg tataaacctg taaagttttt ttcaatttaa 8341 agtagcaaat taatctttat aataaaaaat aacaaaaaag tgttgcttat tattggtatt 8401 attttaatac tagataatag aatttctttt cttttaatct ttcatttatg tgtgcgatgc 8461 aatacaatga attggcggaa atatgtgtag tttccttttc acttaatagt ccttatatta 8521 atatcaaaaa actataaaca aatatttaaa atggtatagc aaattcatca aatcttctta 8581 gttcttttat actatagctt agaatttgta agtaagtcta attttttatt tattttgaac 8641 tcattctgta tctaatgtat atacttatga tgctaaatga cttatttggt catttttata 8701 gccctaacgc tttctattat tactgtttat aaccaactca ttctgaagcc cactgcaggt 8761 cgactctaga ggatccccgg gtaccgagct cccaaatttc tccaaccagt cttcaattat 8821 tctttccaac ttcaaccaat tatatagaaa tgaagaatgg aaaaattcta gtcttcaaca 8881 gtcattcttt aaagacaaac aagcttctat tcaacacaat gaactcttgc aaattattaa 8941 ccaaatctcc ctcccctaca tttctcacaa caatatagtc aagaaatact cttcttcagg 9001 atttttctct tctcctggga atattttact tccatatact attggaacta ttcctctata 9061 gtattctccc atatctctta tatgctcatt atattgcagc attaattgat tttatactta 9121 tatatttttt ttagtattca aacatttatt cactctcttt ttttaatatt taattcgtat 9181 tattatcaat caataatatt ttttattcaa actctactct taaaattcta tcctttacta 9241 tttattccgc ttccacatgc aaaaattctc aattcctgtc attgcatgca aattttatgc 9301 aaaccatttc ttaataccgg tttattatta acatttaggc gccaagaccc agacacgtca 9361 ttatgaagtt aaataaagtt ttttttaata taaaaactga aaggttaagt gttggggtat 9421 gcagaggatg tagaatgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt 9481 atcaacttga aaaagtggca ccgagtcggt gctttttttt tcctaggaag cttgatgcat 9541 agcttgagta ttctatagtg tcacctaaat agcttggcgc tcgaattcac tggccgtcgt 9601 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca 9661 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca 9721 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg 9781 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt 9841 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc 9901 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc 9961 accgtcatca ccgaaacgcg cga //