LOCUS       sequence-151078-        8380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Cas9 and a CRISPR sgRNA that alleviates toxicity to Cas9 in 
            Toxoplasma gondii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8380)
  AUTHORS   Sidik SM, Huet D, Ganesan SM, Huynh MH, Wang T, Nasamu AS, Thiru P, 
            Saeij JP, Carruthers VB, Niles JC, Lourido S
  TITLE     A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential 
            Apicomplexan Genes.
  JOURNAL   Cell. 2016 Sep 8;166(6):1423-1435.e12. doi: 
            10.1016/j.cell.2016.08.019. Epub 2016 Sep 2.
  PUBMED    27594426
REFERENCE   2  (bases 1 to 8380)
  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"; doi: 
            "10.1016/j.cell.2016.08.019"; journalName: "Cell"; date: "2016-09-8-
            8"; volume: "166"; issue: "6"; pages: "1423-1435.e12"
COMMENT     Sequence Label: pU6-Decoy
FEATURES             Location/Qualifiers
     source          1..8380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             514..579
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             586..606
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             631..4731
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             631..4731
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4732..4779
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4807..4824)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4813..5020
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5029..5169
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5029..5158
                     /label=AAV2 ITR
     rep_origin      5244..5699
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5331..5350)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5541..5562
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5716..5735)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5835..5857
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5895..5913)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5981..6085
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6086..6154,6155..6946)
                     /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(6304..6323)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7117..7705
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7606..7625
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_RNA        8288..8363
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ctagaggtac catgcatgtc ccgcgttcgt gaaattctct gcatcagcgg agtgatcagg
       61 aatcatcgtc tcagcgggat gacgttgcgg agcaggccgg ctcgcggtgg gcagtcagat
      121 gccgaaggcg taactcagga cggcttgcgc tcatcgcaga acaggggtgg tgcctgcatt
      181 gggtgcggtt ggtgatcctg gttggaccgg tggagatgcg cgcgcacgaa ggggatgtgt
      241 cagaaacatt ttgtttgttc tctgtgaact tttagatgtg ttaaaggcgg cgaatattag
      301 cagagagtcc tccttgttgg attctctctt gaatttcgcc ctttccttct ctttgcgagt
      361 ctcgtagaga acaagcactc gttcgccgtc cctgacgacg caacccgcgc agaagacatc
      421 caccaaacgg tgttacacaa tcaccttgtg tgaagttctt gcggaaaact actcgttggc
      481 attttttctt gaattccctt tttcgacacc atggactata aggaccacga cggagactac
      541 aaggatcatg atattgatta caaagacgat gacgataaga tggccccaaa gaagaagcgg
      601 aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg cctggacatc
      661 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
      721 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
      781 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
      841 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
      901 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
      961 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
     1021 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
     1081 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
     1141 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
     1201 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
     1261 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
     1321 gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac
     1381 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
     1441 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
     1501 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
     1561 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
     1621 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
     1681 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
     1741 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
     1801 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
     1861 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
     1921 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
     1981 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
     2041 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
     2101 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
     2161 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
     2221 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
     2281 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
     2341 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
     2401 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
     2461 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
     2521 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
     2581 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
     2641 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
     2701 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
     2761 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
     2821 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
     2881 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
     2941 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
     3001 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
     3061 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
     3121 cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc
     3181 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc
     3241 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg
     3301 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
     3361 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
     3421 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg
     3481 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
     3541 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
     3601 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
     3661 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
     3721 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
     3781 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag
     3841 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa
     3901 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg
     3961 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac
     4021 ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
     4081 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
     4141 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
     4201 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
     4261 agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa
     4321 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
     4381 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag
     4441 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg
     4501 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac
     4561 ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc
     4621 gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc
     4681 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga caaaaggccg
     4741 gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagt aagaattcct agagctcgct
     4801 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     4861 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     4921 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     4981 agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag
     5041 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     5101 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     5161 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5221 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5281 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5341 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5401 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     5461 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5521 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5581 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5641 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     5701 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     5761 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5821 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5881 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5941 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6001 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6061 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6121 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6181 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6241 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6301 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6361 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6421 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6481 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6541 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6601 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6661 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6721 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6781 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
     6841 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6901 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6961 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7021 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7081 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7141 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7201 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7261 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7321 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7381 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7441 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7501 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7561 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7621 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7681 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7741 cttttgctgg ccttttgctc acatgggatg agacaaagtg cgcgagttga aatcgtcgtg
     7801 gggacgattt caccgcggcc acatgttgga gacactgagg gcacacggga aacgcgaaag
     7861 atttcaaatt aacgtaccca aacgcgaaag cttgcgcagc atacactcga agcgaacatc
     7921 ccgaaccatc gagaggcgga gagcgataag tctttcacgc tgcgaagtgt tgcgacggct
     7981 gcgccgctgc actgtgaatt gggcgccaat attgcatcct aggcctgacg cgcctcctgc
     8041 agaacgcgag acactgggat atgtagagcc aagggggaaa ccttcgaact ctcgaatgtc
     8101 ttctctgaca agaatcatat ttccatcagt tctgtcagat tttcaaatgg cgacctgcag
     8161 aggcctgctt cctccctgtg cgctcttcga aggggctttc tgtcgcgcag ggtcacctcg
     8221 tccccgaagg gggtgtttgc cttctggtaa atggggatgt caagttgaga atgcagttta
     8281 gcaccgtgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     8341 aaaaagtggc accgagtcgg tgcttttttt ttctttttct
//
