LOCUS       sequence-215181-        9731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9, with FCP promoter/terminator, Golden Gate Assembly, for 
            Phaeodactylum transformation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9731)
  AUTHORS   Graff van Creveld S, Ben-Dor S, Mizrachi A, Alcolombri U, Hopes A, 
            Mock T, Rosenwasser S, Vardi A
  TITLE     Biochemical Characterization of a Novel Redox-Regulated Metacaspase 
            in a Marine Diatom.
  JOURNAL   Front Microbiol. 2021 Sep 8;12:688199. doi: 
            10.3389/fmicb.2021.688199. eCollection 2021.
  PUBMED    34566902
REFERENCE   2  (bases 1 to 9731)
  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: "Front 
            Microbiol."; date: "2021-09-8"; volume: "12:688199. doi"; pages: " 
            10.3389/fmicb.2021.688199. eCollection 2021"
COMMENT     Sequence Label: pICH47742:PtFCP:Cas9YFP
FEATURES             Location/Qualifiers
     source          1..9731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    40..64
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             420..4523
                     /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             420..4523
                     /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             420..4523
                     /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             4536..4556
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4563..4565,4566..4568,4569..5282)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4608..4629)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4869..4888)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5216..5237
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5573..5597
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(5753..6901)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(7237..7825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7317..7336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7996..8787,8788..8856))
                     /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     8619..8638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8857..8961)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9048..9679
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt
       61 aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgggg
      121 tggatgcagt gggccccact ctgtgaagac aagcaagaat tcaagcttgg agacatacct
      181 tcagcgtcgt ctacactgtc acagtcaact gacagtaatc gttgatccgg agagattcaa
      241 aattcaatct gtttggacct ggataagaca caagagcgac atcctgacat gaacgccgta
      301 aacagcaaat cctggttgaa cacgtatcct tttgggggcc tcccgctacg acgctcgctc
      361 cagctggggc ttccttacta tacacagcgc gcatatttca cggttgccag atgtcaagaa
      421 tggacaagaa gtactccatt gggctcgata tcggcacaaa cagcgtcggc tgggccgtca
      481 ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc
      541 acagcataaa gaagaacctc attggcgccc tcctgttcga ctccggggag acggccgaag
      601 ccacgcggct caaaagaaca gcacggcgca gatatacccg cagaaagaat cggatctgct
      661 acctgcagga gatctttagt aatgagatgg ctaaggtgga tgactctttc ttccataggc
      721 tggaggagtc ctttttggtg gaggaggata aaaagcacga gcgccaccca atctttggca
      781 atatcgtgga cgaggtggcg taccatgaaa agtacccaac catatatcat ctgaggaaga
      841 agcttgtaga cagtactgat aaggctgact tgcggttgat ctatctcgcg ctggcgcata
      901 tgatcaaatt tcggggacac ttcctcatcg agggggacct gaacccagac aacagcgatg
      961 tcgacaaact ctttatccaa ctggttcaga cttacaatca gcttttcgaa gagaacccga
     1021 tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc
     1081 ggctcgaaaa cctcatcgca cagctccctg gggagaagaa gaacggcctg tttggtaatc
     1141 ttatcgccct gtcactcggg ctgaccccca actttaaatc taacttcgac ctggccgaag
     1201 atgccaagct tcaactgagc aaagacacct acgatgatga tctcgacaat ctgctggccc
     1261 agatcggcga ccagtacgca gacctttttt tggcggcaaa gaacctgtca gacgccattc
     1321 tgctgagtga tattctgcga gtgaacacgg agatcaccaa agctccgctg agcgctagta
     1381 tgatcaagcg ctatgatgag caccaccaag acttgacttt gctgaaggcc cttgtcagac
     1441 agcaactgcc tgagaagtac aaggaaattt tcttcgatca gtctaaaaat ggctacgccg
     1501 gatacattga cggcggagca agccaggagg aattttacaa atttattaag cccatcttgg
     1561 aaaaaatgga cggcaccgag gagctgctgg taaagcttaa cagagaagat ctgttgcgca
     1621 aacagcgcac tttcgacaat ggaagcatcc cccaccagat tcacctgggc gaactgcacg
     1681 ctatcctcag gcggcaagag gatttctacc cctttttgaa agataacagg gaaaagattg
     1741 agaaaatcct cacatttcgg ataccctact atgtaggccc cctcgcccgg ggaaattcca
     1801 gattcgcgtg gatgactcgc aaatcagaag agactatcac tccctggaac ttcgaggaag
     1861 tcgtggataa gggggcctct gcccagtcct tcatcgaaag gatgactaac tttgataaaa
     1921 atctgcctaa cgaaaaggtg cttcctaaac actctctgct gtacgagtac ttcacagttt
     1981 ataacgagct caccaaggtc aaatacgtca cagaagggat gagaaagcca gcattcctgt
     2041 ctggagagca gaagaaagct atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg
     2101 tgaaacagct caaagaagat tatttcaaaa agattgaatg tttcgactct gttgaaatca
     2161 gcggagtgga ggatcgcttc aacgcatccc tgggaacgta tcacgatctc ctgaaaatca
     2221 ttaaagacaa ggacttcctg gacaatgagg agaacgagga cattcttgag gacattgtcc
     2281 tcacccttac gttgtttgaa gatagggaga tgattgaaga acgcttgaaa acttacgctc
     2341 atctcttcga cgacaaagtc atgaaacagc tcaagaggcg ccgatataca ggatgggggc
     2401 ggctgtcaag aaaactgatc aatgggatcc gagacaagca gagtggaaag acaatcctgg
     2461 attttcttaa gtccgatgga tttgccaacc ggaacttcat gcagttgatc catgatgact
     2521 ctctcacctt taaggaggac atccagaaag cacaagtttc tggccagggg gacagtctcc
     2581 acgagcacat cgctaatctt gcaggtagcc cagctatcaa aaagggaata ctgcagaccg
     2641 ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca taagcccgag aatatcgtta
     2701 tcgagatggc ccgagagaac caaactaccc agaagggaca gaagaacagt agggaaagga
     2761 tgaagaggat tgaagagggt ataaaagaac tggggtccca aatccttaag gaacacccag
     2821 ttgaaaacac ccagcttcag aatgagaagc tctacctgta ctacctgcag aacggcaggg
     2881 acatgtacgt ggatcaggaa ctggacatca atcggctctc cgactacgac gtggatcata
     2941 tcgtgcccca gtcttttctc aaagatgatt ctattgataa taaagtgttg acaagatccg
     3001 ataaaaatag agggaagagt gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa
     3061 attattggcg gcagctgctg aacgccaaac tgatcacaca acggaagttc gataatctga
     3121 ctaaggctga acgaggtggc ctgtctgagt tggataaagc cggcttcatc aaaaggcagc
     3181 ttgttgagac acgccagatc accaagcacg tggcccaaat tctcgattca cgcatgaaca
     3241 ccaagtacga tgaaaatgac aaactgattc gagaggtgaa agttattact ctgaagtcta
     3301 agctggtttc agatttcaga aaggactttc agttttataa ggtgagagag atcaacaatt
     3361 accaccatgc gcatgatgcc tacctgaatg cagtggtagg cactgcactt atcaaaaaat
     3421 atcccaagct tgaatctgaa tttgtttacg gagactataa agtgtacgat gttaggaaaa
     3481 tgatcgcaaa gtctgagcag gaaataggca aggccaccgc taagtacttc ttttacagca
     3541 atattatgaa ttttttcaag accgagatta cactggccaa tggagagatt cggaagcgac
     3601 cacttatcga aacaaacgga gaaacaggag aaatcgtgtg ggacaagggt agggatttcg
     3661 cgacagtccg gaaggtcctg tccatgccgc aggtgaacat cgttaaaaag accgaagtac
     3721 agaccggagg cttctccaag gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg
     3781 cacgcaaaaa agattgggac cccaagaaat acggcggatt cgattctcct acagtcgctt
     3841 acagtgtact ggttgtggcc aaagtggaga aagggaagtc taaaaaactc aaaagcgtca
     3901 aggaactgct gggcatcaca atcatggagc gatcaagctt cgaaaaaaac cccatcgact
     3961 ttctcgaggc gaaaggatat aaagaggtca aaaaagacct catcattaag cttcccaagt
     4021 actctctctt tgagcttgaa aacggccgga aacgaatgct cgctagtgcg ggcgagctgc
     4081 agaaaggtaa cgagctggca ctgccctcta aatacgttaa tttcttgtat ctggccagcc
     4141 actatgaaaa gctcaaagga tctcccgaag ataatgagca gaagcagctg ttcgtggaac
     4201 aacacaaaca ctaccttgat gagatcatcg agcaaataag cgaattctcc aaaagagtga
     4261 tcctcgccga cgctaacctc gataaggtgc tttctgctta caataagcac agggataagc
     4321 ccatcaggga gcaggcagaa aacattatcc acttgtttac tctgaccaac ttgggcgcgc
     4381 ctgcagcctt caagtacttc gacaccacca tagacagaaa gcggtacacc tctacaaagg
     4441 aggtcctgga cgccacactg attcatcagt caattacggg gctctatgaa acaagaatcg
     4501 acctctctca gctcggtgga gacagcaggg ctgaccccaa gaagaagagg aaggtgggtt
     4561 cgatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     4621 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     4681 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     4741 ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
     4801 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     4861 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4921 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4981 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     5041 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     5101 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     5161 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     5221 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     5281 gagcttcctt ccttaaaaat ttaattttca ttagttgcag tcactccgct ttggtttcac
     5341 agtcaggaat aacactagct cgacttcacc atggatgcca atctcgccta ttcatggtgt
     5401 ataaaagttc aacatccaaa gctagaactt ttggaaagag aaagaatatc cgaatagggc
     5461 acggcgtgcc gtattgttgg agtggactag cagaaagtga ggaaggcaca ggatgagttt
     5521 tctcgagcgc tactattgtc ttctgcacga agtggtttaa actatcagtg tttgacagga
     5581 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     5641 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
     5701 cagatcaggc gctggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt
     5761 tgcaatgcac caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact
     5821 cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc
     5881 acatgaggcg gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt
     5941 cgaacatccg tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt
     6001 agtggtcgcc agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg
     6061 ttttcttgcc acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc
     6121 caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg
     6181 ccttcgtgta ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg
     6241 tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca
     6301 ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt
     6361 tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg
     6421 tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg
     6481 caatatcgaa caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg
     6541 cggcctgctt ggcctcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct
     6601 tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct
     6661 gttcaagacg acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca
     6721 gttgcacgct gtcgcgctcg atcttggccg tagcttgctg gaccatcgag ccgacggact
     6781 ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg
     6841 cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca
     6901 ttcaccctcc ttgcgggatt gccccgactc acgccggggc aatgtgccct tattcctgat
     6961 ttgacccgcc tggtgccttg gtgtccagat aatccacctt atcggcaatg aagtcggtcc
     7021 cgtagaccgt ctggccgtcc ttctcgtact tggtattccg aatcttgccc tgcacgaata
     7081 ccagcgaccc cttgcccaaa tacttgccgt gggcctcggc ctgagagcca aaacacttga
     7141 tgcggaagaa gtcggtgcgc tcctgcttgt cgccggcatc gttgcgccac atctaggatc
     7201 tgccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8041 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8101 cttaccatct ggccccagtg ctgcaatgat accgcgagam ccacgctcac cggctccaga
     8161 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8221 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8281 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8341 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8401 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8461 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8521 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8581 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8641 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8701 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8761 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8821 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8881 aagcatttat caaggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8941 taaacaaata ggggttccgc gcacgaattg gccagcgctg ccatttttgg ggtgaggccg
     9001 ttcgcggccg aggggcgcag cccctggggg gatgggaggc ccgcgttagc gggccgggag
     9061 ggttcgagaa gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc
     9121 ctggttaaaa acaaggttta taaatattgg tttaaaagca ggttaaaaga caggttagcg
     9181 gtggccgaaa aacgggcgga aacccttgca aatgctggat tttctgcctg tggacagccc
     9241 ctcaaatgtc aataggtgcg cccctcatct gtcagcactc tgcccctcaa gtgtcaagga
     9301 tcgcgcccct catctgtcag tagtcgcgcc cctcaagtgt caataccgca gggcacttat
     9361 ccccaggctt gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat
     9421 ttgcgaggct ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa
     9481 cgccgcgccg ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag
     9541 ggccaagttt tccgcgaggt atccacaacg ccggcggccg cggtgtctcg cacacggctt
     9601 cgacggcgtt tctggcgcgt ttgcagggcc atagacggcc gccagcccag cggcgagggc
     9661 aaccagcccg gtgagcgtcg caaaggagat cctgatctga ctgatgggct gcctgtatcg
     9721 agtggtgatt t
//
