LOCUS       sequence-320072-        9110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9110)
  AUTHORS   Sreekanth V, Zhou Q, Kokkonda P, Bermudez-Cabrera HC, Lim D, Law BK,
            Holmes BR, Chaudhary SK, Pergu R, Leger BS, Walker JA, Gifford DK, 
            Sherwood RI, Choudhary A
  TITLE     Chemogenetic System Demonstrates That Cas9 Longevity Impacts Genome 
            Editing Outcomes.
  JOURNAL   ACS Cent Sci. 2020 Dec 23;6(12):2228-2237. doi: 
            10.1021/acscentsci.0c00129. Epub 2020 Nov 18.
  PUBMED    33376784
REFERENCE   2  (bases 1 to 9110)
  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/acscentsci.0c00129"; journalName: "ACS Cent Sci"; date: 
            "2020-12-23- 23"; volume: "6"; issue: "12"; pages: "2228-2237"
COMMENT     Sequence Label: pX330 NC-FKBP-Cas9
FEATURES             Location/Qualifiers
     source          1..9110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        268..343
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        440..725
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          962..1189
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1201..1210
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1210..1275
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1282..1302
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1333..1653
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1654..5754
                     /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             1654..5754
                     /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             5755..5802
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5803..6126
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     primer_bind     complement(6151..6168)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6157..6364
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6373..6513
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6373..6502
                     /label=AAV2 ITR
     rep_origin      6588..7043
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6675..6694)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6885..6906
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7060..7079)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7179..7201
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7239..7257)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7325..7429
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7430..7498,7499..8290)
                     /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(7648..7667)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8461..9049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8950..8969
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
      301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
      361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
      421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
      541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
      661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
      781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgaggggcg
      841 gggcggggcg aggcggrgag gkgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      901 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      961 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
     1021 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
     1081 tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt
     1141 attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt
     1201 gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa
     1261 gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca
     1321 gcagccggat ccggagtgca ggtggaaacc atctccccag gagacgggcg caccttcccc
     1381 aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg aaagaaagtt
     1441 gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca ggaggtgatc
     1501 cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa actgactata
     1561 tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc acatgccact
     1621 ctcgtcttcg atgtggagct tctaaaactg gaagacaaga agtacagcat cggcctggac
     1681 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     1741 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     1801 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     1861 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     1921 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     1981 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     2041 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     2101 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     2161 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     2221 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     2281 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     2341 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     2401 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     2461 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     2521 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     2581 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     2641 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     2701 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     2761 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     2821 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     2881 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     2941 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     3001 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     3061 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     3121 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     3181 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     3241 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     3301 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     3361 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     3421 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     3481 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     3541 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     3601 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     3661 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     3721 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     3781 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     3841 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     3901 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     3961 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     4021 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     4081 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     4141 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     4201 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
     4261 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     4321 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     4381 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     4441 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     4501 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     4561 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     4621 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     4681 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     4741 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     4801 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     4861 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     4921 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     4981 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     5041 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     5101 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     5161 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     5221 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     5281 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
     5341 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     5401 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     5461 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     5521 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     5581 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
     5641 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
     5701 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaaagg
     5761 ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agggagtgca ggtggaaacc
     5821 atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac
     5881 accgggatgc ttgaagatgg aaagaaagtt gattcctccc gggacagaaa caagcccttt
     5941 aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg
     6001 agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg tgccactggg
     6061 cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg
     6121 gaataagaat tcctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     6181 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     6241 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     6301 ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg
     6361 gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     6421 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     6481 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     6541 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     6601 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     6661 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     6721 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     6781 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     6841 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     6901 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     6961 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     7021 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     7081 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     7141 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     7201 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     7261 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     7321 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7381 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7441 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7501 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7561 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7621 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7681 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7741 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7801 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7861 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7921 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7981 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     8041 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     8101 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
     8161 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     8221 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8281 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     8341 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8401 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8461 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8521 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8581 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8641 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8701 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     8761 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     8821 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     8881 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     8941 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     9001 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     9061 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
