LOCUS       sequence-149703-        8576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 codon optimized for Phaeodactylum tricornutum and a 
            sgRNA driven by a U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8576)
  AUTHORS   Nymark M, Sharma AK, Sparstad T, Bones AM, Winge P
  TITLE     A CRISPR/Cas9 system adapted for gene editing in marine algae.
  JOURNAL   Sci Rep. 2016 Apr 25;6:24951. doi: 10.1038/srep24951.
  PUBMED    27108533
REFERENCE   2  (bases 1 to 8576)
  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: "Sci Rep."; 
            date: "2016-04-25"; volume: "6:24951. doi"; pages: " 
            10.1038/srep24951"
COMMENT     Sequence Label: pKS diaCas9_sgRNA
FEATURES             Location/Qualifiers
     source          1..8576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1117..1182
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1189..1209
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1234..5334
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1234..5334
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5335..5382
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     misc_RNA        5918..5993
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(6340..6356)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6341..6357)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(6385..6403)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6424..6440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6424..6440)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6437..6459)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6448..6464
                     /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(6472..6478,6479..6496,6497..6502))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6517..6538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6655..6672)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6826..7414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6906..6925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7585..8376,8377..8445))
                     /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     8208..8227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8446..8550)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcaa
      661 tctcgcctat tcatggtgta taaaagttca acatccaaag ctagaacttt tggaaagaga
      721 aagaatatcc gaatagggca cggcgtgccg tattgttgga gtggactagc agaaagtgag
      781 gaaggcacag gatgagtttt ctcgagacac atagcttcag cgtcgtgtag gctaggcaga
      841 ggtgagtttt ctcgagacat accttcagcg tcgtcttcac tgtcacagtc aactgacagt
      901 aatcgttgat ccggagagat tcaaaattca atctgtttgg acctggataa gacacaagag
      961 cgacatcctg acatgaacgc cgtaaacagc aaatcctggt tgaacacgta tccttttggg
     1021 ggcctccgct acgacgctcg ctccagctgg ggcttcctta ctatacacag cgcgcatatt
     1081 tcacggttgc cagaagtcaa gatggcctct agaatggact ataaggacca cgacggagat
     1141 tacaaggatc atgatattga ttacaaagac gatgatgata aaatggcccc aaagaagaag
     1201 cggaaagtcg gtatacacgg agtcccagca gccgacaaga agtactcgat tggattggac
     1261 atcggcacca actctgttgg atgggccgtc atcacggacg agtacaaggt gcccagcaag
     1321 aaattcaaag tactgggaaa cactgaccgc cattcgatca agaagaattt gattggagct
     1381 ttgctgtttg acagcggcga aacagcggaa gccacgcggc ttaaacgaac cgcgagacga
     1441 agatatacgc gacggaagaa ccgtatctgc tatctacaag agattttctc gaatgagatg
     1501 gcaaaagtcg atgactcttt ctttcaccga ttggaagaat ccttcctcgt tgaagaggat
     1561 aagaaacatg agcggcaccc gatatttgga aacattgtgg acgaggtggc ttaccatgag
     1621 aaatacccga ctatttacca cctgcgaaag aaattggtgg actcgacgga caaagcggat
     1681 cttcgcctaa tctatttggc gctagcacat atgattaagt ttcgcggcca cttcttgatc
     1741 gaaggcgact tgaatccgga caacagtgat gttgacaaac tgtttattca acttgtacaa
     1801 acttacaacc agctctttga agaaaatccc attaacgctt cgggagtgga cgcgaaagcc
     1861 atcctttctg cccgactcag caagagccga cgcctagaaa atttgattgc gcaacttccc
     1921 ggcgagaaaa agaatggact ttttggaaat cttattgctt tgtcgctggg ccttacgccg
     1981 aactttaaga gcaacttcga ccttgcggaa gatgccaaat tgcaactgag taaagacaca
     2041 tatgacgacg acctcgacaa tcttttggca caaataggag atcagtacgc cgacctcttt
     2101 ttggcggcta agaacttgtc cgacgccatt ctgttgtcgg acatccttcg agttaatacg
     2161 gagattacta aggcgccctt gagcgcctct atgatcaaaa gatacgacga gcatcaccaa
     2221 gacctgaccc tccttaaagc tctcgtccgg caacagctgc ctgagaagta caaagagatt
     2281 ttctttgatc aaagcaaaaa cggatacgcg ggctacattg acgggggagc ctcgcaagaa
     2341 gagttctaca agtttattaa accgatcctg gaaaagatgg acggcacgga ggaattactc
     2401 gtgaaactta accgagagga ccttttgcgt aagcaacgga ctttcgacaa cggatctatt
     2461 ccgcaccaaa tccatctcgg agaattgcac gcaattctgc gccggcaaga agatttttat
     2521 ccatttctca aggacaaccg ggaaaagatt gagaaaatct taaccttccg cataccctac
     2581 tacgtcggcc ctttggccag gggaaattcg agatttgcct ggatgacgcg aaagagtgag
     2641 gaaactatta ctccctggaa tttcgaagaa gtggtagaca aaggagcttc cgcgcaatcc
     2701 tttatcgagc gcatgaccaa cttcgataag aacctcccga atgagaaagt cctacccaag
     2761 cattcgttgc tctacgaata ctttacggtg tataatgagc tgactaaagt caaatacgtt
     2821 acagagggaa tgcgaaaacc ggctttcctc tcgggagagc agaaaaaggc catagtcgat
     2881 cttctcttta aaacgaaccg taaagttacc gtgaaacaac taaaagagga ctacttcaag
     2941 aaaatcgaat gctttgactc cgttgaaatt tccggcgttg aagatcggtt caatgcctcc
     3001 ttgggaacat accacgatct cctgaaaatt atcaaagaca aggactttct tgacaatgag
     3061 gaaaacgagg acattcttga agatattgtc ttgacgctaa cactttttga ggatcgagag
     3121 atgattgagg aacgccttaa aacctatgcg catctgttcg acgacaaagt tatgaagcaa
     3181 ttgaagcgcc ggagatatac gggatgggga aggttgagtc ggaaactgat aaatggcata
     3241 cgcgataagc agtccggcaa aacaattctc gatttcttga agtccgacgg atttgcgaat
     3301 cgaaacttca tgcaactgat ccatgacgat tcgttgacct ttaaagagga cattcaaaaa
     3361 gcccaagtat ccggacaagg agattctttg cacgagcata ttgcgaatct tgctggaagt
     3421 ccggctatta aaaagggcat tctccagaca gtaaaggtcg tggatgaact cgttaaagtt
     3481 atgggccgac acaaacccga gaacatagtc atcgaaatgg cgagagagaa ccagaccacc
     3541 caaaagggac agaaaaattc acgcgagcga atgaagcgga ttgaagaggg gatcaaagag
     3601 cttggcagcc aaattctgaa agaacacccg gttgaaaaca cccaattgca gaacgagaag
     3661 ctctacctgt actatcttca aaatgggcgt gatatgtacg tcgatcaaga attggacatc
     3721 aaccggctat ccgattacga tgtggaccat attgttcctc agtcgtttct caaagacgac
     3781 tccatagaca ataaggttct aacgagaagc gacaagaacc gcggcaaatc agacaacgtc
     3841 ccctccgagg aagtagttaa gaaaatgaag aattactggc gccaattgct gaacgctaag
     3901 ctcattaccc aaagaaagtt tgacaatttg acgaaagccg agcgaggcgg gctctcggaa
     3961 ctggataagg ccggcttcat taaaagacaa ttggtggaaa cgcgacaaat cacaaagcac
     4021 gtcgcacaga ttctcgactc ccgcatgaac actaagtatg acgagaatga caaactaatc
     4081 cgggaagtca aagtgattac cttgaagtcc aagctggttt ccgattttcg taaagatttt
     4141 cagttttaca aagttcgcga gatcaacaac taccatcacg cgcacgacgc ttacttgaac
     4201 gctgtcgtgg gaacggcctt gattaaaaaa taccctaagc tggaaagcga gttcgtttac
     4261 ggcgattaca aggtgtatga cgtacgcaaa atgatcgcga agtcagagca ggaaattggc
     4321 aaagctactg cgaagtactt cttttatagc aacataatga attttttcaa aacggagatt
     4381 accctcgcta acggcgagat tcgcaagcgc cctcttatcg agacaaacgg cgaaacgggg
     4441 gagattgtct gggataaagg ccgcgatttt gcgactgtgc ggaaagtctt gtcgatgccg
     4501 caagtgaata tagtcaaaaa gacggaggtg caaacaggcg gcttttcgaa agaatctatc
     4561 cttcccaaaa ggaactctga taagttgatt gcgcgaaaaa aggattggga ccctaagaag
     4621 tatggcggct ttgacagccc gaccgtagca tattctgtcc ttgtggtggc caaagtcgaa
     4681 aagggcaaat ccaagaaact gaagagtgtg aaagaattgc ttgggattac gatcatggaa
     4741 agatcatctt ttgagaagaa tcccatcgac tttctcgaag ctaaaggcta caaagaagtg
     4801 aaaaaggacc tgattatcaa gttgcctaag tactccttgt ttgagctgga aaatggccgt
     4861 aagcgaatgc tcgcttctgc cggcgaactc cagaagggaa acgaattggc gctgccctcc
     4921 aaatatgtta actttcttta ccttgcctcg cactatgaga agctcaaggg ctcccccgag
     4981 gataatgagc aaaaacaact gtttgtagaa cagcataagc actacttgga cgagatcatt
     5041 gagcaaatta gcgagttctc caagcgagtc atccttgccg acgctaatct ggacaaagtc
     5101 ctgtccgcct acaataagca ccgcgataag ccaatccgag agcaagcgga gaatattatc
     5161 cacttgttta cgctcaccaa tcttggagcc cctgccgcct ttaagtactt tgacaccacc
     5221 atcgaccgca agaggtacac cagcaccaaa gaagtcttgg acgccaccct cattcaccaa
     5281 tcaatcaccg gcttgtacga gacacgcatc gacctgtctc aactcggagg cgacaaaagg
     5341 ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agtaatctgg atccctacct
     5401 cgactttggc tgggacactt tcagtgagga caagaagctc cagaagcgtg ctatcgaact
     5461 caaccaggga cgtgcggcac aaatgggcat ccttgctctc atggtgcacg aacagttggg
     5521 agtctctatc cttccttaaa aatttaattt tcattagttg cagtcactcc gctttggttt
     5581 cacagtcagg aataacacta gctcgtcttc actgcaggtt ggctcggaag ttggtgttga
     5641 cggtgagctg gaaattggtt gtcggtcact gctagcgaga agaaaacgga ggacagaagg
     5701 aagtgaaact cggttcgttc tcgacagcct cactgtcaat atgctcattt tcaatcctta
     5761 gcgcttttaa tgtcgaattg acggtaaatt gaataggatc tataatatct acaaggtact
     5821 ttgacacgcc aagtattcat tgttagtcaa caatatttta gagctttata aggtcaaaaa
     5881 acaccttcaa agtcgaggag accgagagag ggtctcagtt ttagagctag aaatagcaag
     5941 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
     6001 tagaaccgct cacccatgct atcgtatgtc atttacattg acaaactgct atgaatgcgt
     6061 ttgcgataac ttcagctcgc tgaagcccta gacatttcgt ttgactaaat tgttttccgg
     6121 ttgctttact accgaccggt ctgcgggtag catgtcttgg cttttgcgag ttcgagagaa
     6181 tttatggaca agtgcgttta cagtttggaa cacgagtgtg agtgagcgga attggctatc
     6241 tataacattc ttatttcagg aggctggtga ttagcagaat tttcaaagat ctcgaatcta
     6301 aaatcgccat gggctcgacg aacttttctg caagcttatc gataccgtcg acctcgaggg
     6361 ggggcccggt accagctttt gttcccttta gtgagggtta attccgagct tggcgtaatc
     6421 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6481 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6541 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6601 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6661 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6721 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6781 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6841 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6901 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6961 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7021 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7081 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7141 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7201 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7261 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7321 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7381 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7441 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7501 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7561 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7621 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7681 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagagc cacgctcacc
     7741 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7801 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7861 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7921 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7981 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8041 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8101 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8161 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8221 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8281 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8341 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8401 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8461 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8521 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
