LOCUS       sequence-259456-       10286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-T base editing using LpCDA1L1_1 fused to N terminal of nCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10286)
  AUTHORS   Cheng TL, Li S, Yuan B, Wang X, Zhou W, Qiu Z
  TITLE     Expanding C-T base editing toolkit with diversified cytidine 
            deaminases.
  JOURNAL   Nat Commun. 2019 Aug 9;10(1):3612. doi: 10.1038/s41467-019-11562-6.
  PUBMED    31399578
REFERENCE   2  (bases 1 to 10286)
  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: "2019-08-9"; volume: "10(1):3612. doi"; pages: " 
            10.1038/s41467-019-11562-6"
COMMENT     Sequence Label: N-8-BE
FEATURES             Location/Qualifiers
     source          1..10286
                     /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"
     intron          409..1347
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1304..1324
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(2080..2103,2104..2106,2107..6180)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2080..6180
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6181..6228
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6238..6483
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="NLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAY
                     DESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6496..6516
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6532..6585
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(6586..6588,6589..7299)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6586..6588,6589..7299)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6889..6908)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7236..7257
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7327..7344)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7333..7540
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7549..7689
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7549..7678
                     /label=AAV2 ITR
     rep_origin      7764..8219
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7851..7870)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8061..8082
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8236..8255)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8355..8377
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8415..8433)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8501..8605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8606..8674,8675..9466)
                     /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(8824..8843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9637..10225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10126..10145
                     /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 cgaaacacca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
      301 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
      361 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
      421 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
      481 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
      541 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
      601 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
      661 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
      721 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
      781 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
      841 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
      901 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
      961 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     1021 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     1081 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     1141 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     1201 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     1261 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     1321 gttcaaagtt tttttcttcc atttcaggtg tcgtgaggtg tcgtgagcgc catggccgga
     1381 tccgccggcg acgagaacgt gcgcgtgagc aagaagctgg acttcaacac cttcgagttc
     1441 gagttcgaga acctgcacta cgccgagggc cgcggccgca cctacgtgat cttcgacgtg
     1501 aagccccaga gcgagggcgg ccgcggcgag cgcctgtggg gctacgtgcg caacaacccc
     1561 ctggacgacc acgccgaggt gatcctgatg agcaagatca acgaccacct ggagacacac
     1621 cagggcaact acaccatgac ctggtacatg agctggagcc cctgcggcaa ctgcagcagc
     1681 gagctggtgc cctggctgca gaacctgctg gaggagcagc agcacaccct gaccatgtac
     1741 ttcagccgca tctacgacaa ggaccgcgcc gtggaccacc gcggcctgtg cgacctgcag
     1801 cacgtggtga gcaacggctt ccagatgggc gtgatgggcc agaccgaggt ggacacctgc
     1861 ctggccgagt acgtggaggc cagcggctgc ccccccctga agtggctgca catgaccgac
     1921 agcaacgcca cccagaccca ggacaagctg agcagcatcc tgatgaaccg cttcgtgcgc
     1981 tgcgccggca tgcgcgagag cggcatgccc ctgcacctgt tcaccgctag acgatccagc
     2041 gagactcccg ggacctcaga gtccgccaca ccccgatccg acaagaagta cagcatcggc
     2101 ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     2161 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     2221 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     2281 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     2341 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     2401 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     2461 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     2521 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     2581 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     2641 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     2701 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     2761 ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag cctgggcctg
     2821 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     2881 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     2941 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     3001 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     3061 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     3121 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     3181 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     3241 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     3301 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     3361 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     3421 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     3481 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     3541 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     3601 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     3661 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     3721 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     3781 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     3841 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     3901 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     3961 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     4021 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     4081 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     4141 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     4201 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     4261 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     4321 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     4381 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     4441 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     4501 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     4561 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     4621 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     4681 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     4741 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     4801 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     4861 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     4921 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     4981 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     5041 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     5101 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     5161 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     5221 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     5281 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     5341 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     5401 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     5461 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
     5521 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     5581 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     5641 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     5701 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     5761 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     5821 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     5881 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     5941 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     6001 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     6061 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     6121 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     6181 aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagga attcaytaat
     6241 ctgtcagata ttattgaaaa ggaaaccggc aagcaactgg ttatccagga atccatcctc
     6301 atgctcccag aggaggtgga agaagtcatt gggaacaagc cggaaagcga tatactcgtg
     6361 cacaccgcct acgacgagag caccgacgag aatgtcatgc ttctgactag cgacgcccct
     6421 gaatacaagc cttgggctct ggtcatacag gatagcaacg gtgagaacaa gattaagatg
     6481 ctctctggtg gttctcccaa gaagaagagg aaagtcgcta gcggcagtgg agagggcaga
     6541 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagtgag caagggcgag
     6601 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     6661 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     6721 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     6781 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     6841 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     6901 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     6961 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     7021 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     7081 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     7141 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     7201 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     7261 gccgccggga tcactctcgg catggacgag ctgtacaagg aattctaact agagctcgct
     7321 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     7381 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     7441 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     7501 agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag
     7561 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     7621 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     7681 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     7741 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     7801 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     7861 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     7921 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     7981 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     8041 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     8101 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     8161 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     8221 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     8281 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     8341 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     8401 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     8461 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     8521 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     8581 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     8641 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     8701 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     8761 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     8821 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     8881 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     8941 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     9001 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     9061 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     9121 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     9181 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     9241 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     9301 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
     9361 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     9421 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     9481 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     9541 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     9601 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     9661 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     9721 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     9781 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     9841 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     9901 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     9961 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
    10021 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
    10081 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
    10141 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
    10201 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
    10261 cttttgctgg ccttttgctc acatgt
//
