LOCUS       sequence-259451-       10268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-T base editing using LjCDA1L1_4 fused to N terminal of nCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10268)
  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 10268)
  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-4-BE
FEATURES             Location/Qualifiers
     source          1..10268
                     /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(2062..2085,2086..2088,2089..6162)
                     /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             2062..6162
                     /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             6163..6210
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6220..6465
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="NLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAY
                     DESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6478..6498
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6514..6567
                     /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(6568..6570,6571..7281)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6568..6570,6571..7281)
                     /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(6871..6890)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7218..7239
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7309..7326)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7315..7522
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7531..7671
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7531..7660
                     /label=AAV2 ITR
     rep_origin      7746..8201
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7833..7852)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8043..8064
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8218..8237)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8337..8359
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8397..8415)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8483..8587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8588..8656,8657..9448)
                     /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(8806..8825)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9619..10207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10108..10127
                     /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 gagaagctgg acttcaacac cttcgagttc
     1441 gagttcgaga acctgcacta cgccgagggc cgcggccgca cctacgtgat cttcgacgtg
     1501 aagccccaga gcgagggcgg ccgcggcaag cgcctgtggg gctacgtgcg caacaacccc
     1561 ctggacgacc acgccgaggt gatcctgatg agcaagatca acgaccacct ggaaacccac
     1621 cagggcaact acaccatgac ctggtacatg agctggagcc cctgcggcaa ctgcagcagc
     1681 gagctggtgc cctggctgaa gaacctggag gagcagcagc acaccctgac catgcacttc
     1741 agccgcatct acgacaagga ccgcgccgtg gaccaccgcg gcctgtgcga cctgcagcgc
     1801 gtggtgagca actacttcca gatgggcgtg atgcgcaaga aggaggtgaa gaagtgcctg
     1861 gccgagtacg tggaggccag cggccgcacc ctgcgctggc tgcgcaccac caccagcaac
     1921 gccggccgcc gccgccgcaa gctgtatagc atcctggtgc gctgcgccgg catgcgcgag
     1981 agcggcatgc ccctgcacct gttcaccgct agacgatcca gcgagactcc cgggacctca
     2041 gagtccgcca caccccgatc cgacaagaag tacagcatcg gcctggccat cggcaccaac
     2101 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     2161 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     2221 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     2281 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     2341 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     2401 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     2461 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     2521 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     2581 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     2641 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     2701 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     2761 aatggcctgt tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     2821 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     2881 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     2941 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     3001 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     3061 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     3121 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     3181 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     3241 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     3301 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     3361 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     3421 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     3481 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     3541 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     3601 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     3661 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     3721 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     3781 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     3841 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     3901 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     3961 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     4021 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     4081 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     4141 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     4201 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     4261 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     4321 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     4381 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     4441 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     4501 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     4561 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     4621 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     4681 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     4741 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     4801 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     4861 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     4921 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     4981 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     5041 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     5101 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     5161 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     5221 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     5281 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     5341 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     5401 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     5461 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     5521 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     5581 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     5641 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     5701 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     5761 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     5821 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     5881 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     5941 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     6001 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     6061 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     6121 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg
     6181 aaaaaggccg gccaggcaaa aaagaaaaag gaattcayta atctgtcaga tattattgaa
     6241 aaggaaaccg gcaagcaact ggttatccag gaatccatcc tcatgctccc agaggaggtg
     6301 gaagaagtca ttgggaacaa gccggaaagc gatatactcg tgcacaccgc ctacgacgag
     6361 agcaccgacg agaatgtcat gcttctgact agcgacgccc ctgaatacaa gccttgggct
     6421 ctggtcatac aggatagcaa cggtgagaac aagattaaga tgctctctgg tggttctccc
     6481 aagaagaaga ggaaagtcgc tagcggcagt ggagagggca gaggaagtct gctaacatgc
     6541 ggtgacgtcg aggagaatcc tggcccagtg agcaagggcg aggagctgtt caccggggtg
     6601 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     6661 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     6721 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     6781 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     6841 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     6901 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     6961 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     7021 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     7081 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     7141 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     7201 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     7261 ggcatggacg agctgtacaa ggaattctaa ctagagctcg ctgatcagcc tcgactgtgc
     7321 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     7381 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     7441 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     7501 agaatagcag gcatgctggg gagcggccgc aggaacccct agtgatggag ttggccactc
     7561 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     7621 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     7681 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     7741 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     7801 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     7861 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     7921 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     7981 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     8041 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga
     8101 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     8161 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     8221 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     8281 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     8341 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     8401 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     8461 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     8521 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     8581 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     8641 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     8701 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     8761 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     8821 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     8881 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     8941 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     9001 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     9061 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     9121 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     9181 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     9241 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     9301 tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     9361 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     9421 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     9481 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     9541 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     9601 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     9661 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     9721 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     9781 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     9841 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     9901 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     9961 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
    10021 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
    10081 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
    10141 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
    10201 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
    10261 tcacatgt
//
