LOCUS       sequence-234354-        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     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..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        join(2144..2149,2150..2167,2168..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(join(2814..3527,3528..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"
     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(3630..3656)
                     /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             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"
     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        9435..9510
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(9550..9568)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(9551..9568)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(9587..9604)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9587..9603)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9596..9618)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(9812..9831)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9931..9953
                     /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 gttgggctca
     9421 atttgctcta gaatgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     9481 caacttgaaa aagtggcacc gagtcggtgc tttttttttc ctaggaagct tgatgcatag
     9541 cttgagtatt ctatagtgtc acctaaatag cttggcgctc gaattcactg gccgtcgttt
     9601 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     9661 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     9721 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     9781 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     9841 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     9901 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     9961 cgtcatcacc gaaacgcgcg a
//
