LOCUS       sequence-155393-        9301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Cas9 and chloramphenicol acetyltransferase (CAT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9301)
  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 9301)
  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: pCas9-CAT
FEATURES             Location/Qualifiers
     source          1..9301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             509..574
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             581..601
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             626..4726
                     /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             626..4726
                     /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             4727..4774
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4802..4819)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4808..5015
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5024..5164
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5024..5153
                     /label=AAV2 ITR
     rep_origin      5239..5694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5326..5345)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5536..5557
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5711..5730)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5830..5852
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5890..5908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5976..6080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6081..6149,6150..6941)
                     /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(6299..6318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7112..7700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7601..7620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7773..7789
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7774..7790
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             8305..8967
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MHEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
                     LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFS
                     SLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVAN
                     MDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(8374..8393)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(9285..9301)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9285..9301)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 ggtaccatgc atgtcccgcg ttcgtgaaat tctctgcatc agcggagtga tcaggaatca
       61 tcgtctcagc gggatgacgt tgcggagcag gccggctcgc ggtgggcagt cagatgccga
      121 aggcgtaact caggacggct tgcgctcatc gcagaacagg ggtggtgcct gcattgggtg
      181 cggttggtga tcctggttgg accggtggag atgcgcgcgc acgaagggga tgtgtcagaa
      241 acattttgtt tgttctctgt gaacttttag atgtgttaaa ggcggcgaat attancagag
      301 agtcctcctt gttggattct ctcttgaatt tcgccctttc cttctctttg cgagtctcgt
      361 agagaacaag cactcgttcg ccgtccctga cgacgcaacc cgcgcagaag acatccacca
      421 aacggtgtta cacaatcacc ttgtgtgaag ttcttgcgga aaactactcg ttggcatttt
      481 ttcttgaatt ccctttttcg acaccatgga ctataaggac cacgacggag actacaagga
      541 tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt
      601 cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac
      661 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
      721 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
      781 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
      841 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
      901 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
      961 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     1021 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     1081 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     1141 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     1201 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     1261 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     1321 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     1381 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     1441 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     1501 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     1561 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     1621 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     1681 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     1741 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     1801 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     1861 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     1921 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     1981 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     2041 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     2101 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     2161 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     2221 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     2281 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     2341 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     2401 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     2461 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     2521 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     2581 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     2641 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     2701 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
     2761 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
     2821 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
     2881 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
     2941 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
     3001 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
     3061 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
     3121 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
     3181 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
     3241 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
     3301 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
     3361 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
     3421 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
     3481 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
     3541 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
     3601 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
     3661 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
     3721 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
     3781 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
     3841 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
     3901 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
     3961 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
     4021 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
     4081 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
     4141 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
     4201 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
     4261 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
     4321 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
     4381 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
     4441 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
     4501 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
     4561 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
     4621 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
     4681 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc
     4741 cacgaaaaag gccggccagg caaaaaagaa aaagtaagaa ttcctagagc tcgctgatca
     4801 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4861 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4921 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4981 gaggattggg aagagaatag caggcatgct ggggagcggc cgcaggaacc cctagtgatg
     5041 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     5101 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     5161 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     5221 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5281 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5341 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5401 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     5461 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5521 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5581 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5641 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     5701 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5761 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5821 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5881 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5941 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6001 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6061 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6121 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6181 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6241 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6301 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6361 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6421 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6481 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6541 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6601 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6661 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6721 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6781 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6841 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6901 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6961 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7021 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7081 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7141 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7201 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     7261 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7321 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7381 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7441 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7501 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7561 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7621 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7681 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7741 gctggccttt tgctcacatg tcgggccccc cctcgaggtc gacggtatcg ataagcttga
     7801 tggcgatgca tgtcccgcgt tcgtgaaatt ctctgcatca gcggagtgat caggaatcat
     7861 cgtctcagcg ggatgacgtt gcggagcagg ccggctcgcg gtgggcagtc agatgccgaa
     7921 ggcgtaactc aggacggctt gcgctcatcg cagaacaggg gtggtgcctg cattgggtgc
     7981 ggttggtgat cctggttgga ccggtggaga tgcgcgcgca cgaaggggat gtgtcagaaa
     8041 cattttgttt gttctctgtg aacttttaga tgtgttaaag gcggcgaata ttagcagaga
     8101 gtcctccttg ttggattctc tcttgaattt cgccctttcc ttctctttgc gagtctcgta
     8161 gagaacaagc actcgttcgc cgtccctgac gacgcaaccc gcgcagaaga catccaccaa
     8221 acggtgttac acaatcacct tgtgtgaagt tcttgcggaa aactactcgt tggcattttt
     8281 tcttgaattc cctttttcga caaaatgcat gagaaaaaaa tcactggata taccaccgtt
     8341 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     8401 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     8461 aagcacaagt tttatccggc atttattcac attcttgccc gcctgatgaa tgctcatccg
     8521 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     8581 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     8641 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     8701 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     8761 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     8821 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     8881 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     8941 tgcgatgagt ggcagggcgg ggcttaatta atcaccgttg tgctcacttc tcaaatcgac
     9001 aaaggaaaca cacttcgtgc agcatgtgcc ccattataaa gaaactgagt tgttccgttg
     9061 tggcttgcag gtgtcacatc cacaaaaacc ggccgactct aaataggagt gtttcgcagc
     9121 aagcagcgaa agtttatgac tgggtccgaa tctctgaacg gatgtgtggc ggacctggct
     9181 gatgttgatc gccgtcgaca cacgcgccac atgggtcaat acacaagaca gctatcagtt
     9241 gttttagtcg aaccggttaa cacaattctt gcccccccga gggggatcca ctagttctag
     9301 a
//
