LOCUS       sequence-410971-        8223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for human codon-optimized increased fidelity 
            SpCas9-HF1 (without U6-sgRNA coding sequence, with silent 
            mutations).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8223)
  AUTHORS   Kulcsar PI, Talas A, Toth E, Nyeste A, Ligeti Z, Welker Z, Welker E
  TITLE     Blackjack mutations improve the on-target activities of increased 
            fidelity variants of SpCas9 with 5'G-extended sgRNAs.
  JOURNAL   Nat Commun. 2020 Mar 6;11(1):1223. doi: 10.1038/s41467-020-15021-5.
  PUBMED    32144253
REFERENCE   2  (bases 1 to 8223)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2020-03-6"; volume: "11(1):1223. doi"; pages: " 
            10.1038/s41467-020-15021-5"
COMMENT     Sequence Label: pX330-Flag-SpCas9-HF1 (without sgRNA; with sil...
FEATURES             Location/Qualifiers
     source          1..8223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        294..579
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     rep_origin      complement(651..1239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(731..750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1410..2201,2202..2270))
                     /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     2033..2052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2271..2375)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2443..2461
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2499..2521)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2621..2640
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2657..3112)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2794..2815)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3006..3025
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3187..3327
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3198..3327
                     /label=AAV2 ITR
     polyA_signal    3336..3543
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3532..3549
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3577..3624)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(join(3625..4950,4951..4953,4954..5643,5644..5646,
                     5647..5745,5746..5748,5749..6237,6238..6240,6241..7725))
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Kleinstiver et al., 2016)"
                     /label=SpCas9-HF1
                     /note="carries the mutations N497A, R661A, Q695A, and 
                     Q926A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFMA
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(3625..7725)
                     /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
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFMA
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(7750..7770)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7777..7842)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      7842..7851
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          7863..8090
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc cccccccccc
       61 cccccccccc cccccccccc cccccccccc cccccccccc cycccccccc cccccccccc
      121 ccccccmccc cccccccccc cccccccccc cccccccccn gscnggcscc cccccccccc
      181 ccccccgccc ccatcgctgc acaaaataat taaaaaataa ataaatacaa aattgggggt
      241 ggggaggggg gggagatggg gagagtgaag cagaacgtgg ggctcacctc gaccatggta
      301 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
      361 tgggcacaat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
      421 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
      481 cgtcaatgga aagtccctat tggcgttact attgacgtca atgggcgggg gtcgttgggc
      541 ggtcagccag gcgggccatt taccgtaagt tatgtaacgg gtacctctag catgtgagca
      601 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      661 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      721 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      781 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      841 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      901 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      961 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1021 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1081 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1141 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1201 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1261 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1321 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1381 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1441 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1501 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg gcttccacgc
     1561 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1621 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1681 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1741 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1801 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1861 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1921 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1981 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2041 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2101 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2161 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2221 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2281 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2341 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2401 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2461 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2521 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2581 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     2641 ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     2701 tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca
     2761 aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta
     2821 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2881 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2941 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3001 aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg
     3061 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt
     3121 tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3181 ggcgcccctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
     3241 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     3301 tggccaactc catcactagg ggttcctgcg gccgctcccc agcatgcctg ctattctctt
     3361 cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac
     3421 tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt
     3481 ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca
     3541 cagtcgaggc tgatcagcga gctctaggaa ttcttacttt ttcttttttg cctggccggc
     3601 ctttttcgtg gccgccggcc ttttgtcgcc tcccagctga gacaggtcga tccgtgtctc
     3661 gtacaggccg gtgatgctct ggtggatcag ggtggcgtcc agcacctctt tggtgctggt
     3721 gtacctcttc cggtcgatgg tggtgtcaaa gtacttgaag gcggcagggg ctcccagatt
     3781 ggtcagggta aacaggtgga tgatattctc ggcctgctct ctgatgggct tatcccggtg
     3841 cttgttgtag gcggacagca ctttgtccag attagcgtcg gccaggatca ctctcttgga
     3901 gaactcgctg atctgctcga tgatctcgtc caggtagtgc ttgtgctgtt ccacaaacag
     3961 ctgtttctgc tcattatcct cgggggagcc cttcagcttc tcatagtggc tggccaggta
     4021 caggaagttc acatatttgg agggcagggc cagttcgttt cccttctgca gttcgccggc
     4081 agaggccagc attctcttcc ggccgttttc cagctcgaag agcgagtact taggcagctt
     4141 gatgatcagg tcctttttca cttctttgta gcccttggct tccagaaagt cgatgggatt
     4201 cttctcgaag ctgcttcttt ccatgatggt gatccccagc agctctttca cactcttaag
     4261 cttcttggac ttgccctttt ccactttggc caccaccagc acagaatagg ccacggtggg
     4321 gctgtcgaag ccgccgtact tcttaggatc ccagtccttc tttctggcga tcagcttatc
     4381 gctgttcctc ttgggcagga tagactcttt gctgaagccg cctgtctgca cctcggtctt
     4441 tttcacgata ttcacttggg gcatgctcag cactttccgc acggtggcaa aatcccggcc
     4501 cttatcccac acgatctccc cggtttcgcc gtttgtctcg atcagaggcc gcttccggat
     4561 ctcgccgttg gccagggtaa tctcggtctt gaaaaagttc atgatgttgc tgtagaagaa
     4621 gtacttggcg gtagccttgc cgatttcctg ctcgctcttg gcgatcatct tccgtacgtc
     4681 gtacaccttg tagtcgccgt acacgaactc gctttccagc ttagggtact ttttgatcag
     4741 ggcggttccc acgacggcgt tcaggtacgc gtcgtgggcg tggtggtagt tgttgatctc
     4801 gcgcactttg taaaactgga aatccttccg gaaatcggac accagcttgg acttcagggt
     4861 gatcactttc acttcccgga tcagcttgtc attctcgtcg tacttagtgt tcatccggga
     4921 gtccaggatc tgtgccacgt gctttgtgat cgcccgggtt tccaccagct gtctcttgat
     4981 gaagccggcc ttatccagtt cgctcaggcc gcctctctcg gccttggtca gattgtcgaa
     5041 ctttctctgg gtaatcagct tggcgttcag cagctgccgc cagtagttct tcatcttctt
     5101 cacgacctct tcggagggca cgttgtcgct cttgccccgg ttcttgtcgc ttctggtcag
     5161 caccttgttg tcgatggagt cgtccttaag aaagctctga ggcacgatat ggtccacatc
     5221 gtagtcggac agccggttga tgtccagttc ctggtccacg tacatatccc gcccattctg
     5281 gaggtagtac aggtacagct tctcgttctg gagctgggtg ttttccacgg ggtgttcttt
     5341 caggatctgg ctgcccagct ctttgatgcc ctcttcgatc cgcttcattc tctcgcgact
     5401 gttcttctgt cccttctggg tggtctggtt ctctctggcc atttcgatca cgatgttctc
     5461 gggcttgtgc cggcccatca ctttcaccag ctcgtccacc accttcactg tctggaggat
     5521 gcccttctta atggcggggc tgccggcgag attggcaatg tgctcgtgca ggctatcgcc
     5581 ctggccggac acctgggctt tctggatgtc ctctttaaag gtcaggctgt cgtcgtggat
     5641 cagcgccatg aagtttctgt tggcgaagcc gtcggacttc aggaaatcca ggattgtctt
     5701 gccggactgc ttgtcgcgga tgccgttgat cagcttccgg ctcagcgcgc cccagccggt
     5761 gtatctccgc cgcttcagct gcttcatcac tttgtcgtcg aacaggtggg cataggtttt
     5821 cagccgttcc tcgatcatct ctctgtcctc aaacagtgtc agggtcagca cgatatcttc
     5881 cagaatgtcc tcgttttcct cattgtccag gaagtccttg tccttgataa ttttcagcag
     5941 atcgtggtat gtgcccaggg aggcgttgaa ccgatcttcc acgccggaga tttccacgga
     6001 gtcgaagcac tcgattttct tgaagtagtc ctctttcagc tgcttcacgg tcactttccg
     6061 gttggtcttg aacagcaggt ccacgatggc ctttttctgc tcgccgctca ggaaggcggg
     6121 ctttctcatt ccctcggtca cgtatttcac tttggtcagc tcgttataca cggtgaagta
     6181 ctcgtacagc aggctgtgct tgggcagcac cttctcgttg ggcaggttct tatcgaaggc
     6241 ggtcatccgc tcgatgaagc tctgggcgga agcgcccttg tccaccactt cctcgaagtt
     6301 ccagggggtg atggtttcct cgctctttct ggtcatccag gcgaatctgc tgtttcccct
     6361 ggccagaggg cccacgtagt aggggatgcg gaaggtcagg atcttctcga tcttttcccg
     6421 gttgtccttc aggaatgggt aaaaatcttc ctgccgccgc agaatggcgt gcagctctcc
     6481 caggtggatc tggtggggga tgctgccgtt gtcgaaggtc cgctgcttcc gcagcaggtc
     6541 ctctctgttc agcttcacga gcagttcctc ggtgccgtcc atcttttcca ggatgggctt
     6601 gatgaacttg tagaactctt cctggctggc tccgccgtca atgtagccgg cgtagccgtt
     6661 cttgctctgg tcgaagaaaa tctctttgta cttctcaggc agctgctgcc gcacgagagc
     6721 tttcagcagg gtcaggtcct ggtggtgctc gtcgtatctc ttgatcatag aggcgctcag
     6781 gggggccttg gtgatctcgg tgttcactct caggatgtcg ctcagcagga tggcgtcgga
     6841 caggttcttg gcggccagaa acaggtcggc gtactggtcg ccgatctggg ccagcaggtt
     6901 gtccaggtcg tcgtcgtagg tgtccttgct cagctgcagt ttggcatcct cggccaggtc
     6961 gaagttgctc ttgaagttgg gggtcaggcc caggctcagg gcaatcaggt ttccgaacag
     7021 gccattcttc ttctcgccgg gcagctgggc gatcagattt tccagccgtc tgctcttgct
     7081 cagtctggca gacaggatgg ccttggcgtc cacgccgctg gcgttgatgg ggttttcctc
     7141 gaacagctgg ttgtaggtct gcaccagctg gatgaacagc ttgtccacgt cgctgttgtc
     7201 ggggttcagg tcgccctcga tcaggaagtg gccccggaac ttgatcatgt gggccagggc
     7261 cagatagatc agccgcaggt cggccttgtc ggtgctgtcc accagtttct ttctcaggtg
     7321 gtagatggtg gggtacttct cgtggtaggc cacctcgtcc acgatgttgc cgaagatggg
     7381 gtgccgctcg tgcttcttat cctcttccac caggaaggac tcttccagtc tgtggaagaa
     7441 gctgtcgtcc accttggcca tctcgttgct gaagatctct tgcagatagc agatccggtt
     7501 cttccgtctg gtgtatcttc ttctggcggt tctcttcagc cgggtggcct cggctgtttc
     7561 gccgctgtcg aacagcaggg ctccgatcag gttcttcttg atgctgtgcc ggtcggtgtt
     7621 gcccagcacc ttgaatttct tgctgggcac cttgtactcg tcggtgatca cggcccagcc
     7681 cacagagttg gtgccgatgt ccaggccgat gctgtacttc ttgtcggctg ctgggactcc
     7741 gtggataccg accttccgct tcttctttgg ggccatctta tcgtcatcgt ctttgtaatc
     7801 aatatcatga tccttgtagt ctccgtcgtg gtccttatag tccatggtgg caccggtcca
     7861 acctgaaaaa aagtgatttc aggcaggtgc tccaggtaat taaacattaa taccccacca
     7921 accaaccatc ccttaaaccc ttacctcttg ctcagctaat tacagcccgg aggagaaggg
     7981 ccgtcccgcc cgctcacctg tgggagtaac gcggtcagtc agagccgggg cgggcggcgc
     8041 gaggcggcgg cggagcgggg cacggggcga aggcagcgcg cagcgactcc cgcccgccgc
     8101 gcgcttcgct ttttataggg ccgccgccgc cgccgcctcg ccataaaagg aaactttcgg
     8161 agcgcgccgc tctgattggc tgccgccgca cctctccgcc tcgccccgcc ccgcccctcg
     8221 ccc
//
