LOCUS       sequence-275136-       12386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of ABE6.3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12386)
  AUTHORS   Song CQ, Jiang T, Richter M, Rhym LH, Koblan LW, Zafra MP, Schatoff 
            EM, Doman JL, Cao Y, Dow LE, Zhu LJ, Anderson DG, Liu DR, Yin H, Xue
            W
  TITLE     Adenine base editing in an adult mouse model of tyrosinaemia.
  JOURNAL   Nat Biomed Eng. 2019 Feb 25. pii: 10.1038/s41551-019-0357-8. doi: 
            10.1038/s41551-019-0357-8.
  PUBMED    31740768
REFERENCE   2  (bases 1 to 12386)
  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 Biomed 
            Eng. 2019 Feb 25. pii: 10.1038/s41551-019-0357-8. doi: 
            10.1038/s41551-019-0357-8."
COMMENT     Sequence Label: H446 EF1a RA-ABE6.3 p2A Puro
FEATURES             Location/Qualifiers
     source          1..12386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        69..280
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             307..330
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             337..357
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(382..879,880..975,976..1473,1474..1566)
                     /codon_start=1
                     /product="adenine base editor comprising wild-type E. coli 
                     TadA fused to the evolved TadA*(6.3) mutant (Gaudelli et 
                     al., 2017)"
                     /label=ABE(6.3)
                     /translation="SEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWN
                     RPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFG
                     ARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQS
                     STDSGGSSGGSSGSETPGTSESATPESSGGSSGGSSEVEFSHEYWMRHALTLAKRAWDE
                     REVPVGAVLVLNNRVIGEGWNRSIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTF
                     EPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAA
                     LLCYFFRMRRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGG"
     CDS             join(1567..1590,1591..1593,1594..5667)
                     /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             1567..5667
                     /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             5668..5715
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5731..5787
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5788..6387
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5788..5807)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6284..6304
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6403..6991
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6456..6476)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6874..6885)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7063..7296
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7374..7495
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7411..7430)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7465..7484
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7535..7670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7690..7709)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7691..7709)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7719..7736)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7719..7735)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7728..7750)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7877..8332
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7964..7983)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8174..8195
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8358..8462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8463..8531,8532..9323)
                     /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(8681..8700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9494..10082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9983..10002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10236..10253
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10370..10391
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10406..10411,10412..10429,10430..10436)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10444..10460
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     10449..10471
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10468..10484
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10468..10484
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10503..10523
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10505..10523
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10551..10777
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             10778..10958
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11005..11130
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11623..11856
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12041..12085
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    12269..12386
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgggtttatt acagggacag cagagatcca ctttggcgcc ggctcgagtg gctccggtgc
       61 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      121 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      181 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      241 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac gcgggatccc
      301 accatggatt acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt
      361 atccacggag tcccagcagc ctccgaagtc gagttttccc atgagtactg gatgagacac
      421 gcattgactc tcgcaaagag ggcttgggat gaacgcgagg tgcccgtggg ggcagtactc
      481 gtgcataaca atcgcgtaat cggcgaaggt tggaataggc cgatcggacg ccacgacccc
      541 actgcacatg cggaaatcat ggcccttcga cagggagggc ttgtgatgca gaattatcga
      601 cttatcgatg cgacgctgta cgtcacgctt gaaccttgcg taatgtgcgc gggagctatg
      661 attcactccc gcattggacg agttgtattc ggtgcccgcg acgccaagac gggtgccgca
      721 ggttcactga tggacgtgct gcatcaccca ggcatgaacc accgggtaga aatcacagaa
      781 ggcatattgg cggacgaatg tgcggcgctg ttgtccgact tttttcgcat gcggaggcag
      841 gagatcaagg cccagaaaaa agcacaatcc tctactgact ctggtggttc ttctggtggt
      901 tctagcggca gcgagactcc cgggacctca gagtccgcca cacccgaaag ttctggtggt
      961 tcttctggtg gttcttccga agtcgagttt tcccatgagt actggatgag acacgcattg
     1021 actctcgcaa agagggcttg ggatgaacgc gaggtgcccg tgggggcagt actcgtgctc
     1081 aacaatcgcg taatcggcga aggttggaat aggtcaatcg gactccacga ccccactgca
     1141 catgcggaaa tcatggccct tcgacaggga gggcttgtga tgcagaatta tcgacttatc
     1201 gatgcgacgc tgtacgtcac gtttgaacct tgcgtaatgt gcgcgggagc tatgattcac
     1261 tcccgcattg gacgagttgt attcggtgtt cgcaacgcca agacgggtgc cgcaggttca
     1321 ctgatggacg tgctgcatta cccaggcatg aaccaccggg tagaaatcac agaaggcata
     1381 ttggcggacg aatgtgcggc gctgttgtgt tacttttttc gcatgcggag gcaggtcttt
     1441 aacgcccaga aaaaagcaca atcctctact gactctggtg gttcttctgg tggttctagc
     1501 ggcagcgaga ctcccgggac ctcagagtcc gccacacccg aaagttctgg tggttcttct
     1561 ggtggtgaca agaagtacag catcggcctg gccatcggca ccaactctgt gggctgggcc
     1621 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
     1681 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
     1741 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
     1801 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     1861 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
     1921 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
     1981 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     2041 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     2101 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     2161 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     2221 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcggc
     2281 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     2341 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     2401 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     2461 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     2521 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     2581 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     2641 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     2701 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     2761 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     2821 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     2881 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     2941 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     3001 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     3061 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     3121 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     3181 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     3241 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     3301 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     3361 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     3421 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     3481 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     3541 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     3601 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     3661 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     3721 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     3781 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     3841 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     3901 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     3961 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     4021 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     4081 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
     4141 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     4201 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
     4261 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     4321 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     4381 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
     4441 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     4501 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     4561 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     4621 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     4681 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     4741 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
     4801 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
     4861 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
     4921 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     4981 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     5041 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     5101 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     5161 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     5221 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     5281 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     5341 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     5401 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
     5461 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     5521 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     5581 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     5641 atcgacctgt ctcagctggg aggcgacaag cgtcctgctg ctactaagaa agctggtcaa
     5701 gctaagaaaa agaaagctag cggcagcggc gccaccaact tcagcctgct gaagcaggcc
     5761 ggcgacgtgg aggagaaccc cggccccatg accgagtaca agcccacggt gcgcctcgcc
     5821 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     5881 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     5941 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     6001 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     6061 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     6121 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg
     6181 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     6241 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     6301 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     6361 tgcatgaccc gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc tctggattac
     6421 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     6481 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     6541 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     6601 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     6661 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     6721 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     6781 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     6841 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     6901 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     6961 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     7021 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     7081 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     7141 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     7201 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     7261 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     7321 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     7381 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     7441 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     7501 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     7561 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     7621 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     7681 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     7741 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     7801 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     7861 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     7921 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     7981 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     8041 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     8101 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     8161 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     8221 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     8281 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     8341 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     8401 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     8461 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     8521 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     8581 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     8641 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     8701 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     8761 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     8821 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     8881 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     8941 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     9001 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     9061 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     9121 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     9181 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     9241 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     9301 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     9361 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     9421 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     9481 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     9541 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     9601 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     9661 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     9721 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     9781 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     9841 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     9901 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     9961 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
    10021 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
    10081 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
    10141 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
    10201 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
    10261 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
    10321 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
    10381 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
    10441 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
    10501 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
    10561 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
    10621 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
    10681 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
    10741 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
    10801 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
    10861 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
    10921 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
    10981 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
    11041 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
    11101 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
    11161 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
    11221 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
    11281 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
    11341 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
    11401 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
    11461 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
    11521 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
    11581 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
    11641 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
    11701 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
    11761 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
    11821 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
    11881 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
    11941 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
    12001 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
    12061 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
    12121 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
    12181 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
    12241 cccaaccccg aggggaccca gaccacaatt ttaaaagaaa aggggggatt ggggggtaca
    12301 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
    12361 aacaaattac aaaaattcaa aatttt
//
