LOCUS       sequence-315272-        8833 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 8833)
  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 8833)
  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 N-FKBP-Cas9
FEATURES             Location/Qualifiers
     source          1..8833
                     /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"
     promoter        727..1004
                     /label=chicken beta-actin promoter
     intron          1005..1233
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1245..1254
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1254..1319
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1326..1346
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1377..1697
                     /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             1698..5798
                     /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             1698..5798
                     /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             5799..5846
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5874..5891)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5880..6087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6096..6236
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6096..6225
                     /label=AAV2 ITR
     rep_origin      6311..6766
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6398..6417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6608..6629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6783..6802)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6902..6924
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6962..6980)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7048..7152
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7153..7221,7222..8013)
                     /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(7371..7390)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8184..8772
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8673..8692
                     /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 gcgggggggg
      841 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
      901 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc
     1021 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     1081 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     1141 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac
     1201 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata
     1261 aggaccacga cggagactac aaggatcatg atattgatta caaagacgat gacgataaga
     1321 tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc ggatccggag
     1381 tgcaggtgga aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc ggccagacct
     1441 gcgtggtgca ctacaccggg atgcttgaag atggaaagaa agttgattcc tcccgggaca
     1501 gaaacaagcc ctttaagttt atgctaggca agcaggaggt gatccgaggc tgggaagaag
     1561 gggttgccca gatgagtgtg ggtcagagag ccaaactgac tatatctcca gattatgcct
     1621 atggtgccac tgggcaccca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg
     1681 agcttctaaa actggaagac aagaagtaca gcatcggcct ggacatcggc accaactctg
     1741 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
     1801 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
     1861 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
     1921 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
     1981 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     2041 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     2101 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     2161 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     2221 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     2281 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     2341 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     2401 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     2461 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     2521 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     2581 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     2641 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     2701 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     2761 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     2821 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     2881 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     2941 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     3001 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     3061 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     3121 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     3181 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     3241 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     3301 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     3361 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     3421 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     3481 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     3541 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     3601 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     3661 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     3721 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     3781 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     3841 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     3901 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     3961 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     4021 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     4081 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     4141 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     4201 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     4261 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     4321 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     4381 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     4441 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     4501 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     4561 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     4621 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     4681 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     4741 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     4801 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     4861 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     4921 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     4981 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     5041 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     5101 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     5161 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     5221 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     5281 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     5341 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     5401 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     5461 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     5521 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     5581 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     5641 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     5701 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     5761 acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa
     5821 aggccggcca ggcaaaaaag aaaaagtaag aattcctaga gctcgctgat cagcctcgac
     5881 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     5941 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     6001 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     6061 ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc
     6121 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     6181 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     6241 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     6301 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6361 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6421 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6481 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     6541 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6601 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc gggctattct
     6661 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     6721 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     6781 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     6841 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6901 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     6961 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     7021 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     7081 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     7141 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     7201 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     7261 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     7321 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     7381 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     7441 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     7501 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7561 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7621 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7681 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7741 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7801 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7861 gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7921 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7981 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     8041 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     8101 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     8161 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     8221 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     8281 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     8341 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     8401 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     8461 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     8521 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     8581 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     8641 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8701 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8761 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8821 tttgctcaca tgt
//
