LOCUS       sequence-238880-        9981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 fused with GFP and a sgRNA targeting the 
            Cryptosporidium parvum TK gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9981)
  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 9981)
  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: "Cell Host 
            Microbe. 2019 Jun 18. pii: S1931-3128(19)30252-5. doi: 
            10.1016/j.chom.2019.05.007."
COMMENT     Sequence Label: pACT1:Cas9-GFP, U6:sgTK
FEATURES             Location/Qualifiers
     source          1..9981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..71
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     265..287
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     279..296
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     280..296
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        315..333
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     315..332
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_RNA        complement(373..448)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             2114..6217
                     /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"
     CDS             2114..6217
                     /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"
     CDS             6221..6247
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6227..6253
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             6266..6286
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6293..6313
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6353..6355,6356..7069)
                     /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     7003..7024
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7661..7677)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7661..7677)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7674..7696)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7685..7701
                     /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)."
     promoter        complement(join(7709..7715,7716..7733,7734..7739))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7754..7775
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7892..7909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8063..8651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8143..8162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8822..9613,9614..9682))
                     /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     9445..9464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9683..9787)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9855..9873
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9911..9933)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
       61 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
      121 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
      181 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
      241 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
      301 tcgagcgcca agctatttag gtgacactat agaatactca agctatgcat caagcttcct
      361 aggaaaaaaa aagcaccgac tcggtgccac tttttcaagt tgataacgga ctagccttat
      421 tttaacttgc tatttctagc tctaaaacat tctagagcaa attgagccca acacttaacc
      481 tttcagtttt tatattaaaa aaaactttat ttaacttcat aatgacgtgt ctgggtcttg
      541 gcgcctaaat gttaataata aaccggtatt aagaaatggt ttgcataaaa tttgcatgca
      601 atgacaggaa ttgagaattt ttgcatgtgg aagcggaata aatagtaaag gatagaattt
      661 taagagtaga gtttgaataa aaaatattat tgattgataa taatacgaat taaatattaa
      721 aaaaagagag tgaataaatg tttgaatact aaaaaaaata tataagtata aaatcaatta
      781 atgctgcaat ataatgagca tataagagat atgggagaat actatagagg aatagttcca
      841 atagtatatg gaagtaaaat attcccagga gaagagaaaa atcctgaaga agagtatttc
      901 ttgactatat tgttgtgaga aatgtagggg agggagattt ggttaataat ttgcaagagt
      961 tcattgtgtt gaatagaagc ttgtttgtct ttaaagaatg actgttgaag actagaattt
     1021 ttccattctt catttctata taattggttg aagttggaaa gaataattga agactggttg
     1081 gagaaatttg ggagctcggt acccggggat cctctagagt cgacctgcag tgggcttcag
     1141 aatgagttgg ttataaacag taataataga aagcgttagg gctataaaaa tgaccaaata
     1201 agtcatttag catcataagt atatacatta gatacagaat gagttcaaaa taaataaaaa
     1261 attagactta cttacaaatt ctaagctata gtataaaaga actaagaaga tttgatgaat
     1321 ttgctatacc attttaaata tttgtttata gttttttgat attaatataa ggactattaa
     1381 gtgaaaagga aactacacat atttccgcca attcattgta ttgcatcgca cacataaatg
     1441 aaagattaaa agaaaagaaa ttctattatc tagtattaaa ataataccaa taataagcaa
     1501 cacttttttg ttatttttta ttataaagat taatttgcta ctttaaattg aaaaaaactt
     1561 tacaggttta tacaaattcg tacatcacat gtatacaatg ctacatacat tacttgataa
     1621 atattacgac gtgaaacgta ttaatttaaa cgaatcttag ttatgtaaat aaaaaaaggt
     1681 tagattttag taaaaaaaaa agaccaaatt ttatattaaa tataaatctt gcattgaatg
     1741 catttatgtc aaattgcatg caccttgccg cccaccatgc atagacgcgc gccaatacgt
     1801 attaatagac gattacatga caaagtttag agattaaata aaaatcaagt ataaatttag
     1861 aatagacaac atgtaaaaag tgagatgaat ctttttacga tacgtacagt gcgcattaaa
     1921 aatccaaagt atatttttat tttcctgcgt gcgtatacat atctttaaat acttggataa
     1981 agagataata agaattgtac ggcaagcaaa gtagtagttt aatagagcag tttttatatt
     2041 ttatatatag cttacttaca atatagattt ttttttttca tagatttcgg gcatattaca
     2101 caagtaacaa aagatggaca agaagtacag catcggcctg gacatcggca ccaactctgt
     2161 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
     2221 caacaccgac cggcacagca tcaagaagaa cctgatcggc gccctgctgt tcgacagcgg
     2281 agaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
     2341 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
     2401 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
     2461 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
     2521 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
     2581 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
     2641 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
     2701 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
     2761 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
     2821 cctgttcggc aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     2881 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     2941 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     3001 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     3061 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     3121 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     3181 gaacggctac gccggctaca tcgatggcgg agccagccag gaagagttct acaagttcat
     3241 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     3301 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     3361 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     3421 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     3481 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     3541 gaacttcgag gaagtggtgg acaagggcgc cagcgcccag agcttcatcg agcggatgac
     3601 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     3661 gtacttcacc gtgtacaacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     3721 gcccgccttc ctgagcggcg agcagaaaaa agccatcgtg gacctgctgt tcaagaccaa
     3781 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     3841 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     3901 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     3961 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     4021 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     4081 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     4141 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     4201 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     4261 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     4321 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     4381 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     4441 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     4501 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     4561 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     4621 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg ataacaaagt
     4681 gctgactcgg agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     4741 gaagaagatg aagaactact ggcgccagct gctgaatgcc aagctgatta cccagaggaa
     4801 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     4861 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     4921 ctcccggatg aacactaagt acgacgagaa cgacaaactg atccgggaag tgaaagtgat
     4981 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     5041 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     5101 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     5161 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     5221 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     5281 gatccggaag cggcctctga tcgagacaaa cggcgaaaca ggcgagatcg tgtgggataa
     5341 gggccgggac tttgccaccg tgcggaaagt gctgtctatg ccccaagtga atatcgtgaa
     5401 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     5461 cgacaagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
     5521 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     5581 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     5641 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     5701 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     5761 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     5821 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
     5881 gctgtttgtg gaacagcaca aacactacct ggacgagatc atcgagcaga tcagcgagtt
     5941 ctccaagaga gtgatcctgg ccgacgctaa tctggacaag gtgctgagcg cctacaacaa
     6001 gcacagagac aagcctatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
     6061 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
     6121 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     6181 cgagacacgg atcgacctgt ctcagctggg aggcgacgcc tatccctatg acgtgcccga
     6241 ttatgccagc ctgggcagcg gctcccccaa gaaaaaacgc aaggtggaag atcctaagaa
     6301 aaagcggaaa gtggacggca ttggtagtgg gagcaacggc agcagcggat ccgtgagcaa
     6361 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     6421 cggccacaag ttcagcgtgc gcggcgaggg cgagggcgat gccaccaacg gcaagctgac
     6481 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     6541 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     6601 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatctcct tcaaggacga
     6661 cggcacctac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     6721 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     6781 caacttcaac agccacaacg tctatatcac ggccgacaag cagaagaacg gcatcaaggc
     6841 gaacttcaag atccgccaca acgtcgagga cggcagcgtg cagctcgccg accactacca
     6901 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     6961 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     7021 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaga tcaatattgt
     7081 tcttggagtt gttcttaaca gcttatttcc tgaacacaga tacgtattgc aacttaaata
     7141 gaaatatgaa ttacacaagt tatcacttct tgtagacaat ttcttattac attattcaga
     7201 tctcagttct attctcattt gaaagctaaa aaaaaagatg ataaattgtc taatcaatgc
     7261 taataaactc taaatttagc aagaaaattc tattataatt gttagtcaat gataactaat
     7321 attatggata gaaggtgggc tagtttgaat caaaatagtt actaagtaat gtgtagttcg
     7381 atacgatttt ttccttcttt ttaaacgaaa gtgtttaaat tatgtgcgtg ttgttttcct
     7441 gagcacagta ttagaaatta attaagttga tttgtcaggt gatgtttatt accaaagttc
     7501 aagtccgaaa atagagctgc catttaacaa taaaaaagag cggtccaaat ttaggcaaaa
     7561 caaatttttt aaaaaaagct ccaagtccgg gaattgaacc cgggtcatcc gcgtgacagg
     7621 cggatatact atgcaggcat gcaagcttgg cataatcatg gtcatagctg tttcctgtgt
     7681 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7741 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7801 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7861 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7921 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7981 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     8041 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     8101 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8161 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8221 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8281 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8341 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8401 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8461 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     8521 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8581 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8641 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8701 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8761 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8821 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8881 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8941 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     9001 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     9061 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9121 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9181 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9241 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9301 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9361 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9421 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9481 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9541 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9601 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9661 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9721 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9781 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     9841 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     9901 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     9961 atgccgggag cagacaagcc c
//
