LOCUS       sequence-202024-        9976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9-Nlux and sgRNA. Contains CEN/ARS. No selection 
            marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9976)
  AUTHORS   Poliner E, Takeuchi T, Du ZY, Benning C, Farre EM
  TITLE     Nontransgenic Marker-Free Gene Disruption by an Episomal CRISPR 
            System in the Oleaginous Microalga, Nannochloropsis oceanica 
            CCMP1779.
  JOURNAL   ACS Synth Biol. 2018 Apr 20;7(4):962-968. doi: 
            10.1021/acssynbio.7b00362. Epub 2018 Mar 14.
  PUBMED    29518315
REFERENCE   2  (bases 1 to 9976)
  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"; doi: 
            "10.1021/acssynbio.7b00362"; journalName: "ACS Synth Biol"; date: 
            "2018-04-20- 20"; volume: "7"; issue: "4"; pages: "962-968"
COMMENT     Sequence Label: pNOC-CRISPR-compact
FEATURES             Location/Qualifiers
     source          1..9976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(127..195,196..987)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(345..364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1158..1746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1647..1666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1900..1917
                     /label=L4440
                     /note="L4440 vector, forward primer"
     regulatory      2162..2171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2852..2872)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      2877..2886
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2888..2914)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3467..7567)
                     /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             complement(3467..7567)
                     /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             complement(3467..7567)
                     /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             complement(7568..7588)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7809..7820)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        8811..8886
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           8891..8958
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
ORIGIN
        1 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
       61 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      301 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      361 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      421 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      541 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
      661 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
      721 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
      781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
      841 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
      901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
      961 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1021 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1081 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1141 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1201 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1261 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1321 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1381 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1441 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1501 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1561 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1621 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1681 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1741 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     1801 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     1861 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     1921 gctggtcact ggattttggt tttaggaatt agaaatttta ttgatagaag tattttacaa
     1981 atacaaatac atactaaggg tttcttatat gctcaacaca tgagcgaaac cctataagaa
     2041 ccctaattcc cttatctggg aactactcac acattattct ggagaaaaat agagagagat
     2101 agatttgtag agagagactg gtgatttttg cggactccgg tcggcatcta ctactagtgc
     2161 ggccgccatg gcctctagtg gatcagcttg catgcctgca ggagacgata tcacctcttc
     2221 tgtttccacg ataaaaatag actgctcatt tcttcgtcgt cttcatcgtc tgctttttct
     2281 gcttcgcctc tgtctggggt ctgaaaccac tacacacaca cacaacactc gtactcccac
     2341 tttcacaaaa gcgtaagctc accggctttt cttacacgta cattttagtg gatcccatca
     2401 cgccactacc acgcccgcgg gggatggaac ggaggggaga gagagaaggg ggaagcatgg
     2461 atgaatgaga cattgaggga aaggagggga gggagcagtc catcagggcg ctacctctct
     2521 tgtcccccaa accctgttga gccgttcaac atgtttcatg tttcctccct ccccccttcc
     2581 ctccctggcc tttccgcgga gccattcaag tgacgtctgg accgcaccgt aacaaaatcg
     2641 tttctatggg gggtttgttt gacaaccacg tcttcagcgt ttttaaaaaa aaaagcgggc
     2701 aagccctctc accctcactc atgcccatcc tcctcctctc ctgcggaaca ttcttacaaa
     2761 aggcgtaact cgacgacaac tcaaagaacg acaaacatca atcccaaaaa aaaaatctct
     2821 actcgtctct cttggatctt tccaattgtc agaccttcct cttctttttc gggctagcca
     2881 ccatggacgc gtagtcgggc acgtcgtagg ggtatccggc caggatgcgc tcgcacaggc
     2941 gccagccggt taccccgttg atggtcacgc ggaacaacag cgagccatcc gggttgatca
     3001 gccgctcgtc aatgatctta ttgccgttcc acagggtgcc ggtcaccgtg atcttcttgc
     3061 cgtcgaacac ggcaatgccc tcgtaaggac ggccgaagta gtcaatcatg ttcggcgtca
     3121 caccgtcgat gaccagcgtc ccgtagtgca gtatcacctt gaagtggtga tcgtccacgg
     3181 ggtacacaac cttgaagatc ttctcgatct ggcccatttg gtccccgctc aggccctcgt
     3241 agggaatgat gacgtggatg tcgatcttca ggccgttctc gccgctcagg acgatgcgct
     3301 ggattggggt cacggagacg cccaggttct ggaaaagcga ggacacaccg ccctgctcca
     3361 gcacctggtc gaggttatag ccagctgtct gccgccagtc gcccacgaag tcctcgagag
     3421 taaacaccat gttaacggac ccgctgccgg acccgtcagc cctgctgtct ccaccgagct
     3481 gagagaggtc gattcttgtt tcatagagcc ccgtaattga ctgatgaatc agtgtggcgt
     3541 ccaggacctc ctttgtagag gtgtaccgct ttctgtctat ggtggtgtcg aagtacttga
     3601 aggctgcagg cgcgcccaag ttggtcagag taaacaagtg gataatgttt tctgcctgct
     3661 ccctgatggg cttatccctg tgcttattgt aagcagaaag caccttatcg aggttagcgt
     3721 cggcgaggat cactcttttg gagaattcgc ttatttgctc gatgatctca tcaaggtagt
     3781 gtttgtgttg ttccacgaac agctgcttct gctcattatc ttcgggagac cctttgagct
     3841 tttcatagtg gctggccaga tacaagaaat taacgtattt agagggcagt gccagctcgt
     3901 tacctttctg cagctcgccc gcactagcga gcattcgttt ccggccgttt tcaagctcaa
     3961 agagagagta cttgggaagc ttaatgatga ggtctttttt gacctcttta tatcctttcg
     4021 cctcgagaaa gtcgatgggg tttttttcga agcttgatcg ctccatgatt gtgatgccca
     4081 gcagttcctt gacgcttttg agttttttag acttcccttt ctccactttg gccacaacca
     4141 gtacactgta agcgactgta ggagaatcga atccgccgta tttcttgggg tcccaatctt
     4201 ttttgcgtgc gatcagcttg tcgctgttcc ttttcgggag gatactttcc ttggagaagc
     4261 ctccggtctg tacttcggtc tttttaacga tgttcacctg cggcatggac aggaccttcc
     4321 ggactgtcgc gaaatcccta cccttgtccc acacgatttc tcctgtttct ccgtttgttt
     4381 cgataagtgg tcgcttccga atctctccat tggccagtgt aatctcggtc ttgaaaaaat
     4441 tcataatatt gctgtaaaag aagtacttag cggtggcctt gcctatttcc tgctcagact
     4501 ttgcgatcat tttcctaaca tcgtacactt tatagtctcc gtaaacaaat tcagattcaa
     4561 gcttgggata ttttttgata agtgcagtgc ctaccactgc attcaggtag gcatcatgcg
     4621 catggtggta attgttgatc tctctcacct tataaaactg aaagtccttt ctgaaatctg
     4681 agaccagctt agacttcaga gtaataactt tcacctctcg aatcagtttg tcattttcat
     4741 cgtacttggt gttcatgcgt gaatcgagaa tttgggccac gtgcttggtg atctggcgtg
     4801 tctcaacaag ctgccttttg atgaagccgg ctttatccaa ctcagacagg ccacctcgtt
     4861 cagccttagt cagattatcg aacttccgtt gtgtgatcag tttggcgttc agcagctgcc
     4921 gccaataatt tttcattttc ttgacaactt cttctgaggg gacgttatca ctcttccctc
     4981 tatttttatc ggatcttgtc aacactttat tatcaataga atcatctttg agaaaagact
     5041 ggggcacgat atgatccacg tcgtagtcgg agagccgatt gatgtccagt tcctgatcca
     5101 cgtacatgtc cctgccgttc tgcaggtagt acaggtagag cttctcattc tgaagctggg
     5161 tgttttcaac tgggtgttcc ttaaggattt gggaccccag ttcttttata ccctcttcaa
     5221 tcctcttcat cctttcccta ctgttcttct gtcccttctg ggtagtttgg ttctctcggg
     5281 ccatctcgat aacgatattc tcgggcttat gccttcccat tactttgacg agttcatcca
     5341 cgaccttaac ggtctgcagt attccctttt tgatagctgg gctacctgca agattagcga
     5401 tgtgctcgtg aagactgtcc ccctggccag aaacttgtgc tttctggatg tcctccttaa
     5461 aggtgagaga gtcatcatgg atcaactgca tgaagttccg gttggcaaat ccatcggact
     5521 taagaaaatc caggattgtc tttccactct gcttgtctcg gatcccattg atcagttttc
     5581 ttgacagccg cccccatcct gtatatcggc gcctcttgag ctgtttcatg actttgtcgt
     5641 cgaagagatg agcgtaagtt ttcaagcgtt cttcaatcat ctccctatct tcaaacaacg
     5701 taagggtgag gacaatgtcc tcaagaatgt cctcgttctc ctcattgtcc aggaagtcct
     5761 tgtctttaat gattttcagg agatcgtgat acgttcccag ggatgcgttg aagcgatcct
     5821 ccactccgct gatttcaaca gagtcgaaac attcaatctt tttgaaatag tcttctttga
     5881 gctgtttcac ggtaactttc cggttcgtct tgaagaggag gtccacgata gctttcttct
     5941 gctctccaga caggaatgct ggctttctca tcccttctgt gacgtatttg accttggtga
     6001 gctcgttata aactgtgaag tactcgtaca gcagagagtg tttaggaagc accttttcgt
     6061 taggcagatt tttatcaaag ttagtcatcc tttcgatgaa ggactgggca gaggccccct
     6121 tatccacgac ttcctcgaag ttccagggag tgatggtctc ttctgatttg cgagtcatcc
     6181 acgcgaatct ggaatttccc cgggcgaggg ggcctacata gtagggtatc cgaaatgtga
     6241 ggattttctc aatcttttcc ctgttatctt tcaaaaaggg gtagaaatcc tcttgccgcc
     6301 tgaggatagc gtgcagttcg cccaggtgaa tctggtgggg gatgcttcca ttgtcgaaag
     6361 tgcgctgttt gcgcaacaga tcttctctgt taagctttac cagcagctcc tcggtgccgt
     6421 ccattttttc caagatgggc ttaataaatt tgtaaaattc ctcctggctt gctccgccgt
     6481 caatgtatcc ggcgtagcca tttttagact gatcgaagaa aatttccttg tacttctcag
     6541 gcagttgctg tctgacaagg gccttcagca aagtcaagtc ttggtggtgc tcatcatagc
     6601 gcttgatcat actagcgctc agcggagctt tggtgatctc cgtgttcact cgcagaatat
     6661 cactcagcag aatggcgtct gacaggttct ttgccgccaa aaaaaggtct gcgtactggt
     6721 cgccgatctg ggccagcaga ttgtcgagat catcatcgta ggtgtctttg ctcagttgaa
     6781 gcttggcatc ttcggccagg tcgaagttag atttaaagtt gggggtcagc ccgagtgaca
     6841 gggcgataag attaccaaac aggccgttct tcttctcccc agggagctgt gcgatgaggt
     6901 tttcgagccg ccgggatttg gacagcctag cgctcaggat tgctttggcg tcaactccgg
     6961 atgcgttgat cgggttctct tcgaaaagct gattgtaagt ctgaaccagt tggataaaga
     7021 gtttgtcgac atcgctgttg tctgggttca ggtccccctc gatgaggaag tgtccccgaa
     7081 atttgatcat atgcgccagc gcgagataga tcaaccgcaa gtcagcctta tcagtactgt
     7141 ctacaagctt cttcctcaga tgatatatgg ttgggtactt ttcatggtac gccacctcgt
     7201 ccacgatatt gccaaagatt gggtggcgct cgtgcttttt atcctcctcc accaaaaagg
     7261 actcctccag cctatggaag aaagagtcat ccaccttagc catctcatta ctaaagatct
     7321 cctgcaggta gcagatccga ttctttctgc gggtatatct gcgccgtgct gttcttttga
     7381 gccgcgtggc ttcggccgtc tccccggagt cgaacaggag ggcgccaatg aggttcttct
     7441 ttatgctgtg gcgatcggta ttgcccagaa ctttgaattt tttgctcggc accttgtact
     7501 cgtccgtaat gacggcccag ccgacgctgt ttgtgccgat atcgagccca atggagtact
     7561 tcttgtccac cttccgcttt ttcttgggca tgctcgaggg ttgcgtgtgt atctgtgtgc
     7621 agtggatatt gttaccgagt ttggtgagcg tgagtccgtt agtgccctgg tggtggtgga
     7681 ttaggagagt gggtgactcg gtgtccatgg ctttcttcgc tcattatagg aggggaaagg
     7741 aatgagggag ggtggggaga ccgcgtctgt tgttgaccac cgatttactt cttgcctccc
     7801 ttcccctccc tcccctcaat ccgtacgaca caaatagtag ccgagtgtct gctgcagagc
     7861 gcatgattag tgtggtagac aacgagggag ggaaggatgt acagggcatg gcacggagaa
     7921 gcgatggtgg ccaggaagag gagaggtcgc gagaacagga tgtgttgcga atggataaaa
     7981 acagaaagcg atggctctgg gcttcgaaag caggggacat taggacgtgt agaccatctc
     8041 gacggatccc tctgtatctc tgttgtgcgt gaatgttttc tgtgcacgtg tagtgtgtga
     8101 gagtagaacc cgggaactcg aacagagaaa agcatgggtg gctgtggtgt ggaggcttcg
     8161 ttcccaccac atgcccttct ccttcgcctc gcctctccct gccttcttcc acgcaccctt
     8221 gcgcccctcg ttctcaatac ctggctcact tccaccattc aaacaaccat cacgatacag
     8281 gcatttatct atcgttgaag acttcttcct ccggtagatc ttagccaagg taagaagagg
     8341 ggcatgcagc aaggagaaag aaatgatgca tgatgaggaa tagaagggga ggagggaggg
     8401 atatgatggg aagcgaaagc gcatattctg gtggtctgct gcctgatggg gacgcgtcta
     8461 gctgtgacac tgaggacggt ggctgctggt ggctgcgggc gctgccttgg tgatcaatgg
     8521 gagtaaaggg agggaaggag gtagcgtgaa cggatgacgc ggagaagttt aggggtctct
     8581 ttacgtatcg cccctgccgc ccgcctctct gcgataaatg tgcctgttac cctgcagcct
     8641 ctattcttca ctgtgttcct gttttccaac agcctctatt cttccctgtc ttttgttgca
     8701 gtggcgtcat cctctctttg ccccagtcgt cgttctctcg actcactcac tccccccctc
     8761 cttccctccc tccatccaca gaatcgatga agagcaggcc ttggctcttc gttttagagc
     8821 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     8881 cggtgctttt ggccggcatg gtcccagcct cctcgctggc gccggctggg caacatgctt
     8941 cggcatggcg aatgggactc ctgggtacca tgggaaagaa aggatgagaa aggagaaagg
     9001 acatctagat aaccggcata tcacggtggt gtattagtgt agaatagtga agagaagact
     9061 tgggaaaatg tgtaggaaag gttgtttctg tgtatgtggg ttgggatggg tggctgtttg
     9121 agaaggaaca gcgggcaggg cgatgtagtg ctgaacgggc acggaaccac tgagactgaa
     9181 ggaagtaggg agagagaggg gcaggggacg tgcactttaa tctttgcctc ggtagagtat
     9241 acccatgcaa gagtatgtgg ccacctgtgg tggcttttgg ccaggtctgg tgcagtgcca
     9301 atcatctccc atcaataata caacttcaga acaacggcgc attgatgggg agagagaaag
     9361 taaatttaag taaggggtac gtagtagagg attcaactga aatattttcg aggagcggtt
     9421 gggaagttga ccgattgaaa ggagaaggga ggggagcagg tgtgatagtc atgtgtaaag
     9481 taattctttt ttgccgtcgt cacacaatcc acatcaatga taaaatatgt ttaaggatca
     9541 atcacacgga gtcggtcata aggcaaccgc aaacgcaatg caaactagca agcaagcagg
     9601 accacaacaa caaccattac catcacaggc gacagcagca gcagctgcag cagcagcaag
     9661 gcaagcaaag gccatctgcg cgtggacttt tccaggcacc ggctttaagc gtaattttca
     9721 atgattgttg tccgtgtatc ctctcaccct ttcaccgctt gcgcgcacgt gagcagcaca
     9781 actggtaatg cccgaggaca atagacgagg aaaaagaaga agaaaaatga agagggagtg
     9841 atgaagaaga agagaagaat aaaagaagac agaaaagaag ataaaaaagt caaaagacga
     9901 caaagacgga aaaaaaagaa acgcgcaatt ttaacaccag caacgacgac gaaaagggcg
     9961 tcagctttgc accggt
//
