LOCUS       sequence-462135-        9325 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 protein fused with octopine-type VirD2-originating 
            NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9325)
  AUTHORS   Le TT, Choi HI, Kim JW, Yun JH, Lee YH, Jeon EJ, Kwon KK, Cho DH, 
            Choi DY, Park SB, Yoon HR, Lee J, Sim EJ, Lee YJ, Kim HS
  TITLE     Cas9-mediated gene-editing frequency in microalgae is doubled by 
            harnessing the interaction between importin alpha and 
            phytopathogenic NLSs.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Mar 11;122(10):e2415072122. doi: 
            10.1073/pnas.2415072122. Epub 2025 Mar 3.
  PUBMED    40030016
REFERENCE   2  (bases 1 to 9325)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-03-11"; volume: "122(10):e2415072122. 
            doi"; pages: " 10.1073/pnas.2415072122. Epub 2025 Mar 3"
COMMENT     Sequence Label: pET-Octo_VirD2_NLS-Cas9-6xHis
FEATURES             Location/Qualifiers
     source          1..9325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     RBS             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             137..4237
                     /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             137..4237
                     /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             4238..4255
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4274..4318
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4356..4374)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4370..4417
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4454..4909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4541..4560)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4751..4772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(5024..5815,5816..5884))
                     /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     5647..5666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5885..5977)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6058..6646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6547..6566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6800..6817
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6832..6971
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6897..6916)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7057..7079
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7073..7264)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7836..8918)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     8880..8899
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8919..8996)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     9202..9221
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(9309..9325,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(9309..9325,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgctgcaga aacgacgtaa tgacgaggag gcaggtccga gcggagcaaa
      121 ccgtaaagga ttgaaggata agaaatactc aataggctta gatatcggca caaatagcgt
      181 cggatgggcg gtgatcactg atgaatataa ggttccgtct aaaaagttca aggttctggg
      241 aaatacagac cgccacagta tcaaaaaaaa tcttataggg gctcttttat ttgacagtgg
      301 agagacagcg gaagcgactc gtctcaaacg gacagctcgt agaaggtata cacgtcggaa
      361 gaatcgtatt tgttatctac aggagatttt ttcaaatgag atggcgaaag tagatgatag
      421 tttctttcat cgacttgaag agtctttttt ggtggaagaa gacaagaagc atgaacgtca
      481 tcctattttt ggaaatatag tagatgaagt tgcttatcat gagaaatatc caactatcta
      541 tcatctgcga aaaaaattgg tagattctac tgataaagcg gatttgcgct taatctattt
      601 ggccttagcg catatgatta agtttcgtgg tcattttttg attgagggag atttaaatcc
      661 tgataatagt gatgtggaca aactatttat ccagttggta caaacctaca atcaattatt
      721 tgaagaaaac cctattaacg caagtggagt agatgctaaa gcgattcttt ctgcacgatt
      781 gagtaaatca agacgattag aaaatctcat tgctcagctc cccggtgaga agaaaaatgg
      841 cttatttggg aatctcattg ctttgtcatt gggtttgacc cctaatttta aatcaaattt
      901 tgatttggca gaagatgcta aattacagct ttcaaaagat acttacgatg atgatttaga
      961 taatttattg gcgcaaattg gagatcaata tgctgatttg tttttggcag ctaagaattt
     1021 atcagatgct attttacttt cagatatcct aagagtaaat actgaaataa ctaaggctcc
     1081 cctatcagct tcaatgatta aacgctacga tgaacatcat caagacttga ctcttttaaa
     1141 agctttagtt cgacaacaac ttccagaaaa gtataaagaa atcttttttg atcaatcaaa
     1201 aaacggatat gcaggttata ttgatggggg agctagccaa gaagaatttt ataaatttat
     1261 caaaccaatt ttagaaaaaa tggatggtac tgaggaatta ttggtgaaac taaatcgtga
     1321 agatttgctg cgcaagcaac ggacctttga caacggctct attccccatc aaattcactt
     1381 gggtgagctg catgctattt tgagaagaca agaagacttt tatccatttt taaaagacaa
     1441 tcgtgagaag attgaaaaaa tcttgacttt tcgaattcct tattatgttg gtccattggc
     1501 gcgtggcaat agtcgttttg catggatgac tcggaagtct gaagaaacaa ttaccccatg
     1561 gaattttgaa gaagttgtcg ataaaggtgc ttcagctcaa tcatttattg aacgcatgac
     1621 aaactttgat aaaaatcttc caaatgaaaa agtactacca aaacatagtt tgctttatga
     1681 gtattttacg gtttataacg aattgacaaa ggtcaaatat gttactgaag gaatgcgaaa
     1741 accagcattt ctttcaggtg aacagaagaa agccattgtt gatttactct tcaaaacaaa
     1801 tcgaaaagta accgttaagc aattaaaaga agattatttc aaaaaaatag aatgttttga
     1861 tagtgttgaa atttcaggag ttgaagatag atttaatgct tcattaggta cctaccatga
     1921 tttgctaaaa attattaaag ataaagattt tttggataat gaagaaaatg aagatatctt
     1981 agaggatatt gttttaacat tgaccttatt tgaagatagg gagatgattg aggaaagact
     2041 taaaacatat gctcacctct ttgatgataa ggtgatgaaa cagcttaaac gtcgccgtta
     2101 tactggttgg ggacgtttgt ctcgaaaatt gattaatggt attagggata agcaatctgg
     2161 caaaacaata ttagattttt tgaaatcaga tggttttgcc aatcgcaatt ttatgcagct
     2221 gatccatgat gatagtttga catttaaaga agacattcaa aaagcacaag tgtctggaca
     2281 aggcgatagt ttacatgaac atattgcaaa tttagctggt agccctgcta ttaaaaaagg
     2341 tattttacag actgtaaaag ttgttgatga attggtcaaa gtaatggggc ggcataagcc
     2401 agaaaatatc gttattgaaa tggcacgtga aaatcagaca actcaaaagg gccagaaaaa
     2461 ttcgcgagag cgtatgaaac gaatcgaaga aggtatcaaa gaattaggaa gtcagattct
     2521 taaagagcat cctgttgaaa atactcaatt gcaaaatgaa aagctctatc tctattatct
     2581 ccaaaatgga agagacatgt atgtggacca agaattagat attaatcgtt taagtgatta
     2641 tgatgtcgat cacattgttc cacaaagttt ccttaaagac gattcaatag acaataaggt
     2701 cttaacgcgt tctgataaaa atcgtggtaa atcggataac gttccaagtg aagaagtagt
     2761 caaaaagatg aaaaactatt ggagacaact tctaaacgcc aagttaatca ctcaacgtaa
     2821 gtttgataat ttaacgaaag ctgaacgtgg aggtttgagt gaacttgata aagctggttt
     2881 tatcaaacgc caattggttg aaactcgcca aatcactaag catgtggcac aaattttgga
     2941 tagtcgcatg aatactaaat acgatgaaaa tgataaactt attcgagagg ttaaagtgat
     3001 taccttaaaa tctaaattag tttctgactt ccgaaaagat ttccaattct ataaagtacg
     3061 tgagattaac aattaccatc atgcccatga tgcgtatcta aatgccgtcg ttggaactgc
     3121 tttgattaag aaatatccaa aacttgaatc ggagtttgtc tatggtgatt ataaagttta
     3181 tgatgttcgt aaaatgattg ctaagtctga gcaagaaata ggcaaagcaa ccgcaaaata
     3241 tttcttttac tctaatatca tgaacttctt caaaacagaa attacacttg caaatggaga
     3301 gattcgcaaa cgccctctaa tcgaaactaa tggggaaact ggagaaattg tctgggataa
     3361 agggcgagat tttgccacag tgcgcaaagt attgtccatg ccccaagtca atattgtcaa
     3421 gaaaacagaa gtacagacag gcggattctc caaggagtca attttaccaa aaagaaattc
     3481 ggacaagctt attgctcgta aaaaagactg ggatccaaaa aaatatggtg gttttgatag
     3541 tccaacggta gcttattcag tcctagtggt tgctaaggtg gaaaaaggga aatcgaagaa
     3601 gttaaaatcc gttaaagagt tactagggat cacaattatg gaaagaagtt cctttgaaaa
     3661 aaatccgatt gactttttag aagctaaagg atataaggaa gttaaaaaag acttaatcat
     3721 taaactacct aaatatagtc tttttgagtt agaaaacggt cgtaaacgga tgctggctag
     3781 tgccggagaa ttacaaaaag gaaatgagct ggctctgcca agcaaatatg tgaatttttt
     3841 atatttagct agtcattatg aaaagttgaa gggtagtcca gaagataacg aacaaaaaca
     3901 attgtttgtt gagcagcata agcattattt agatgagatt attgagcaaa tcagtgaatt
     3961 ttctaagcgt gttattttag cagatgccaa tttagataaa gttcttagtg catataacaa
     4021 acatagagac aaaccaatac gtgaacaagc agaaaatatt attcatttat ttacgttgac
     4081 gaatcttgga gctcccgctg cttttaaata ttttgataca acaattgatc gtaaacgata
     4141 tacgtctaca aaagaagttt tagatgccac tcttatccat caatccatca ctggtcttta
     4201 tgaaacacgc attgatttga gtcagctagg aggtgaccat caccaccacc atcactaatg
     4261 actcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc
     4321 tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac tagcataacc
     4381 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     4441 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4501 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4561 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4621 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4681 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4741 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4801 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4861 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc tggcggcacg
     4921 atggcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4981 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5041 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5101 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5161 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5221 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5281 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5341 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5401 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5461 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5521 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5581 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5641 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5701 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5761 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5821 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5881 tcatactctt cctttttcaa tcatgattga agcatttatc agggttattg tctcatgagc
     5941 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gtcatgacca aaatccctta
     6001 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6061 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6121 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6181 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     6241 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6301 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6361 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6421 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6481 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6541 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6601 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6661 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6721 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6781 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     6841 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
     6901 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     6961 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     7021 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     7081 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     7141 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     7201 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     7261 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     7321 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     7381 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     7441 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     7501 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     7561 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     7621 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     7681 gccagcctag ccgggtcctc aacgacagga gcacgatcat gctagtcatg ccccgcgccc
     7741 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     7801 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7861 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7921 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     7981 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     8041 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     8101 tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     8161 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     8221 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     8281 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     8341 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     8401 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     8461 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     8521 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     8581 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     8641 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     8701 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     8761 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     8821 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     8881 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     8941 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     9001 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     9061 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     9121 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     9181 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     9241 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc
     9301 cgcgaaatta atacgactca ctata
//
