LOCUS       sequence-171568-        8880 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SpCas9 fused to DHFR domain on both N- and C-termini in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8880)
  AUTHORS   Maji B, Moore CL, Zetsche B, Volz SE, Zhang F, Shoulders MD, 
            Choudhary A
  TITLE     Multidimensional chemical control of CRISPR-Cas9.
  JOURNAL   Nat Chem Biol. 2017 Jan;13(1):9-11. doi: 10.1038/nchembio.2224. Epub
            2016 Oct 31.
  PUBMED    27820801
REFERENCE   2  (bases 1 to 8880)
  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: 
            "10.1038/nchembio.2224"; journalName: "Nat Chem Biol"; date: 
            "2017-01"; volume: "13"; issue: "1"; pages: "9-11"
COMMENT     Sequence Label: DHFR-SpCas9-DHFR
FEATURES             Location/Qualifiers
     source          1..8880
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      224..233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             233..298
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             305..325
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             356..829
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             848..4948
                     /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             848..4948
                     /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             4949..4996
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4997..5473
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDHVIGMETVMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     primer_bind     complement(5498..5515)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5504..5711
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5720..5860
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5720..5849
                     /label=AAV2 ITR
     rep_origin      5935..6390
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6022..6041)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6232..6253
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6407..6426)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6526..6548
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6586..6604)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6672..6776
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6777..6845,6846..7637)
                     /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(6995..7014)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7808..8396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8297..8316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8458..8706
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8458..8478
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8629..8648
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8725..8800
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
       61 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      121 tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc
      181 tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa
      241 ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg acgataagat
      301 ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg gatccatcag
      361 tctgattgcg gcgttagcgg tagattacgt tatcggcatg gaaaacgcca tgccgtggaa
      421 cctgcctgcc gatctcgcct ggtttaaacg caacacctta aataaacccg tgattatggg
      481 ccgccatacc tgggaatcaa tcggtcgtcc gttgccagga cgcaaaaata ttatcctcag
      541 cagtcaaccg agtacggacg atcgcgtaac gtgggtgaag tcggtggatg aagccatcgc
      601 ggcgtgtggt gacgtaccag aaatcatggt gattggcggc ggtcgcgtta ttgaacagtt
      661 cttgccaaaa gcgcaaaaac tgtatctgac gcatatcgac gcagaagtgg aaggcgacac
      721 ccatttcccg gattacgagc cggatgactg ggaatcggta ttcagcgaat tccacgatgc
      781 tgatgcgcag aactctcaca gctattgctt tgagattctg gagcggcgag gaagcggaag
      841 cggatccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
      901 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
      961 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
     1021 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     1081 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     1141 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     1201 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     1261 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     1321 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     1381 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     1441 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     1501 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     1561 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     1621 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     1681 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     1741 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     1801 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     1861 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     1921 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     1981 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     2041 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     2101 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     2161 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     2221 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     2281 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     2341 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     2401 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     2461 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     2521 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     2581 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     2641 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     2701 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     2761 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     2821 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     2881 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     2941 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     3001 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     3061 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     3121 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     3181 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     3241 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     3301 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     3361 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     3421 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     3481 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     3541 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     3601 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     3661 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     3721 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     3781 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     3841 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     3901 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     3961 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     4021 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     4081 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     4141 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     4201 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     4261 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     4321 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     4381 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     4441 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     4501 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     4561 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     4621 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     4681 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     4741 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     4801 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     4861 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     4921 gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca
     4981 ggcaaaaaag aaaaagatca gtctgattgc ggcgttagcg gtagatcacg ttatcggcat
     5041 ggaaaccgtc atgccgtgga acctgcctgc cgatctcgcc tggtttaaac gcaacacctt
     5101 aaataaaccc gtgattatgg gccgccatac ctgggaatca atcggtcgtc cgttgccagg
     5161 acgcaaaaat attatcctca gcagtcaacc gagtacggac gatcgcgtaa cgtgggtgaa
     5221 gtcggtggat gaagccatcg cggcgtgtgg tgacgtacca gaaatcatgg ttattggcgg
     5281 cggtcgcgtt tatgaacagt tcttgccaaa agcgcaaaaa ctgtatctga cgcatatcga
     5341 cgcagaagtg gaaggcgaca cccatttccc ggattacgag ccggatgact gggaatcggt
     5401 attcagcgaa ttccacgatg ctgatgcgca gaactctcac agctattgct ttgagattct
     5461 ggagcggcga taagaattcc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     5521 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     5581 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     5641 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg
     5701 catgctgggg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     5761 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     5821 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     5881 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     5941 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     6001 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     6061 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6121 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     6181 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     6241 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     6301 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6361 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     6421 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     6481 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     6541 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     6601 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6661 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6721 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6781 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6841 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6901 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6961 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     7021 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     7081 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     7141 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7201 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7261 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7321 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7381 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7441 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg
     7501 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7561 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7621 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7681 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7741 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7801 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7861 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7921 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7981 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     8041 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     8101 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8161 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8221 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8281 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8341 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8401 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtgag
     8461 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     8521 attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     8581 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     8641 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     8701 aacaccgggt cttcgagaag acctgtttta gagctagaaa tagcaagtta aaataaggct
     8761 agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt tagagctaga
     8821 aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa
//
