LOCUS       sequence-238877-        9983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 fused with GFP and a sgRNA targeting the 
            Cryptosporidium parvum UPRT gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9983)
  AUTHORS   Wilke G, Funkhouser-Jones LJ, Wang Y, Ravindran S, Wang Q, Beatty 
            WL, Baldridge MT, VanDussen KL, Shen B, Kuhlenschmidt MS, 
            Kuhlenschmidt TB, Witola WH, Stappenbeck TS, Sibley LD
  TITLE     A Stem-Cell-Derived Platform Enables Complete Cryptosporidium 
            Development In Vitro and Genetic Tractability.
  JOURNAL   Cell Host Microbe. 2019 Jun 18. pii: S1931-3128(19)30252-5. doi: 
            10.1016/j.chom.2019.05.007.
  PUBMED    31231046
REFERENCE   2  (bases 1 to 9983)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 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:sgUPRT
FEATURES             Location/Qualifiers
     source          1..9983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..37
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     231..253
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     245..262
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     246..262
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        281..299
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     281..298
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_RNA        complement(339..414)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             2082..6185
                     /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             2082..6185
                     /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             6189..6215
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6195..6221
                     /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             6234..6254
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6261..6281
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6321..6323,6324..7037)
                     /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     6971..6992
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7629..7645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7629..7645)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7642..7664)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7653..7669
                     /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(7677..7683,7684..7701,7702..7707))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7722..7743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7860..7877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8031..8619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8111..8130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8790..9581,9582..9650))
                     /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     9413..9432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9651..9755)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9823..9841
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9879..9901)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
       61 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
      121 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      181 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      241 acgttgtaaa acgacggcca gtgaattcga gcgccaagct atttaggtga cactatagaa
      301 tactcaagct atgcatcaag cttcctagga aaaaaaaagc accgactcgg tgccactttt
      361 tcaagttgat aacggactag ccttatttta acttgctatt tctagctcta aaacattcta
      421 catcctctgc ataccccaac acttaacctt tcagttttta tattaaaaaa aactttattt
      481 aacttcataa tgacgtgtct gggtcttggc gcctaaatgt taataataaa ccggtattaa
      541 gaaatggttt gcataaaatt tgcatgcaat gacaggaatt gagaattttt gcatgtggaa
      601 gcggaataaa tagtaaagga tagaatttta agagtagagt ttgaataaaa aatattattg
      661 attgataata atacgaatta aatattaaaa aaagagagtg aataaatgtt tgaatactaa
      721 aaaaaatata taagtataaa atcaattaat gctgcaatat aatgagcata taagagatat
      781 gggagaatac tatagaggaa tagttccaat agtatatgga agtaaaatat tcccaggaga
      841 agagaaaaat cctgaagaag agtatttctt gactatattg ttgtgagaaa tgtaggggag
      901 ggagatttgg ttaataattt gcaagagttc attgtgttga atagaagctt gtttgtcttt
      961 aaagaatgac tgttgaagac tagaattttt ccattcttca tttctatata attggttgaa
     1021 gttggaaaga ataattgaag actggttgga gaaatttggg agctcggtac ccggggatcc
     1081 tctagagtcg acctgcagtg ggcttcagaa tgagttggtt ataaacagta ataatagaaa
     1141 gcgttagggc tataaaaatg accaaataag tcatttagca tcataagtat atacattaga
     1201 tacagaatga gttcaaaata aataaaaaat tagacttact tacaaattct aagctatagt
     1261 ataaaagaac taagaagatt tgatgaattt gctataccat tttaaatatt tgtttatagt
     1321 tttttgatat taatataagg actattaagt gaaaaggaaa ctacacatat ttccgccaat
     1381 tcattgtatt gcatcgcaca cataaatgaa agattaaaag aaaagaaatt ctattatcta
     1441 gtattaaaat aataccaata ataagcaaca cttttttgtt attttttatt ataaagatta
     1501 atttgctact ttaaattgaa aaaaacttta caggtttata caaattcgta catcacatgt
     1561 atacaatgct acatacatta cttgataaat attacgacgt gaaacgtatt aatttaaacg
     1621 aatcttagtt atgtaaataa aaaaaggtta gattttagta aaaaaaaaag accaaatttt
     1681 atattaaata taaatcttgc attgaatgca tttatgtcaa attgcatgca ccttgccgcc
     1741 caccatgcat agacgcgcgc caatacgtat taatagacga ttacatgaca aagtttagag
     1801 attaaataaa aatcaagtat aaatttagaa tagacaacat gtaaaaagtg agatgaatct
     1861 ttttacgata cgtacagtgc gcattaaaaa tccaaagtat atttttattt tcctgcgtgc
     1921 gtatacatat ctttaaatac ttggataaag agataataag aattgtacgg caagcaaagt
     1981 agtagtttaa tagagcagtt tttatatttt atatatagct tacttacaat atagattttt
     2041 ttttttcata gatttcgggc atattacaca agtaacaaaa gatggacaag aagtacagca
     2101 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     2161 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     2221 tgatcggcgc cctgctgttc gacagcggag aaacagccga ggccacccgg ctgaagagaa
     2281 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     2341 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     2401 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     2461 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     2521 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     2581 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     2641 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     2701 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     2761 cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg
     2821 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     2881 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     2941 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     3001 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     3061 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     3121 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatc gatggcggag
     3181 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     3241 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     3301 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     3361 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     3421 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     3481 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgcca
     3541 gcgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     3601 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag ctgaccaaag
     3661 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaaag
     3721 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     3781 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     3841 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     3901 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     3961 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     4021 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     4081 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     4141 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     4201 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     4261 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     4321 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     4381 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     4441 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     4501 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     4561 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
     4621 tgaaggacga ctccatcgat aacaaagtgc tgactcggag cgacaagaac cggggcaaga
     4681 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cgccagctgc
     4741 tgaatgccaa gctgattacc cagaggaagt tcgacaatct gaccaaggcc gagagaggcg
     4801 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     4861 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaacg
     4921 acaaactgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     4981 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     5041 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     5101 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     5161 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     5221 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     5281 gcgaaacagg cgagatcgtg tgggataagg gccgggactt tgccaccgtg cggaaagtgc
     5341 tgtctatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     5401 aagagtctat cctgcccaag aggaacagcg acaagctgat cgccagaaag aaggactggg
     5461 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     5521 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     5581 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     5641 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     5701 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     5761 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     5821 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaaa cactacctgg
     5881 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     5941 tggacaaggt gctgagcgcc tacaacaagc acagagacaa gcctatcaga gagcaggccg
     6001 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     6061 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     6121 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     6181 gcgacgccta tccctatgac gtgcccgatt atgccagcct gggcagcggc tcccccaaga
     6241 aaaaacgcaa ggtggaagat cctaagaaaa agcggaaagt ggacggcatt ggtagtggga
     6301 gcaacggcag cagcggatcc gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     6361 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgcgc ggcgagggcg
     6421 agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     6481 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     6541 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     6601 aggagcgcac catctccttc aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt
     6661 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     6721 gcaacatcct ggggcacaag ctggagtaca acttcaacag ccacaacgtc tatatcacgg
     6781 ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac gtcgaggacg
     6841 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     6901 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
     6961 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     7021 acgagctgta caagtagatc aatattgttc ttggagttgt tcttaacagc ttatttcctg
     7081 aacacagata cgtattgcaa cttaaataga aatatgaatt acacaagtta tcacttcttg
     7141 tagacaattt cttattacat tattcagatc tcagttctat tctcatttga aagctaaaaa
     7201 aaaagatgat aaattgtcta atcaatgcta ataaactcta aatttagcaa gaaaattcta
     7261 ttataattgt tagtcaatga taactaatat tatggataga aggtgggcta gtttgaatca
     7321 aaatagttac taagtaatgt gtagttcgat acgatttttt ccttcttttt aaacgaaagt
     7381 gtttaaatta tgtgcgtgtt gttttcctga gcacagtatt agaaattaat taagttgatt
     7441 tgtcaggtga tgtttattac caaagttcaa gtccgaaaat agagctgcca tttaacaata
     7501 aaaaagagcg gtccaaattt aggcaaaaca aattttttaa aaaaagctcc aagtccggga
     7561 attgaacccg ggtcatccgc gtgacaggcg gatatactat gcaggcatgc aagcttggca
     7621 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     7681 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     7741 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     7801 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7861 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7921 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7981 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     8041 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     8101 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     8161 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     8221 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     8281 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     8341 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     8401 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     8461 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     8521 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     8581 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     8641 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     8701 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     8761 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     8821 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     8881 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     8941 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     9001 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     9061 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     9121 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     9181 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     9241 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     9301 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     9361 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     9421 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     9481 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     9541 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     9601 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     9661 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     9721 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     9781 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     9841 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     9901 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     9961 tcagcgggtg ttggcgggtg tcg
//
