LOCUS       sequence-215665-       12301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus for constitutive expression of ABE7.10 (not codon 
            optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12301)
  TITLE     Dow Lab Unpublished Base Editors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12301)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLenti-ABE7.10-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..12301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(652..4050,4051..4053,4054..4650,4651,4652..4654,
                     4655..4656,4657..4659,4660..4752)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /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             join(652..675,676..678,679..4752)
                     /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             4765..4785
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4801..4857
                     /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             4858..5457
                     /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(4858..4877)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5354..5374
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5473..6061
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5526..5546)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5944..5955)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6133..6366
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6444..6565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6481..6500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6535..6554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6605..6740
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6760..6779)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6761..6779)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6789..6806)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6789..6805)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6798..6820)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6947..7402
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7034..7053)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7244..7265
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7428..7532
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7533..7601,7602..8393)
                     /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(7751..7770)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8564..9152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9053..9072
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9306..9323
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9440..9461
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9476..9481,9482..9499,9500..9506)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9514..9530
                     /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     9519..9541
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9538..9554
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9538..9554
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9573..9593
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9575..9593
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9621..9847
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9848..10028
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10075..10200
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10693..10926
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11111..11155
                     /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    11339..11456
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        11525..11736
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             join(11762..12262,12263..12301,1..57,58..555,556..651)
                     /codon_start=1
                     /product="adenine base editor comprising wild-type E. coli 
                     TadA fused to the evolved TadA*(7.10) mutant (Gaudelli et 
                     al., 2017)"
                     /label=ABE(7.10)
                     /translation="MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGW
                     NRPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVF
                     GARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQ
                     SSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSSEVEFSHEYWMRHALTLAKRARD
                     EREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVT
                     FEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECA
                     ALLCYFFRMPRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGS"
ORIGIN
        1 cccgggacct cagagtccgc cacacccgaa agttctggtg gttcttctgg tggttcttcc
       61 gaagtcgagt tttcccatga gtactggatg agacacgcat tgactctcgc aaagagggct
      121 cgagatgaac gcgaggtgcc cgtgggggca gtactcgtgc tcaacaatcg cgtaatcggc
      181 gaaggttgga atagggcaat cggactccac gaccccactg cacatgcgga aatcatggcc
      241 cttcgacagg gagggcttgt gatgcagaat tatcgactta tcgatgcgac gctgtacgtc
      301 acgtttgaac cttgcgtaat gtgcgcggga gctatgattc actcccgcat tggacgagtt
      361 gtattcggtg ttcgcaacgc caagacgggt gccgcaggtt cactgatgga cgtgctgcat
      421 tacccaggca tgaaccaccg ggtagaaatc acagaaggca tattggcgga cgaatgtgcg
      481 gcgctgttgt gttacttttt tcgcatgccc aggcaggtct ttaacgccca gaaaaaagca
      541 caatcctcta ctgactctgg tggttcttct ggtggttcta gcggcagcga gactcccggg
      601 acctcagagt ccgccacacc cgaaagttct ggtggttctt ctggtggttc tgataaaaag
      661 tattctattg gtttagccat cggcactaat tccgttggat gggctgtcat aaccgatgaa
      721 tacaaagtac cttcaaagaa atttaaggtg ttggggaaca cagaccgtca ttcgattaaa
      781 aagaatctta tcggtgccct cctattcgat agtggcgaaa cggcagaggc gactcgcctg
      841 aaacgaaccg ctcggagaag gtatacacgt cgcaagaacc gaatatgtta cttacaagaa
      901 atttttagca atgagatggc caaagttgac gattctttct ttcaccgttt ggaagagtcc
      961 ttccttgtcg aagaggacaa gaaacatgaa cggcacccca tctttggaaa catagtagat
     1021 gaggtggcat atcatgaaaa gtacccaacg atttatcacc tcagaaaaaa gctagttgac
     1081 tcaactgata aagcggacct gaggttaatc tacttggctc ttgcccatat gataaagttc
     1141 cgtgggcact ttctcattga gggtgatcta aatccggaca actcggatgt cgacaaactg
     1201 ttcatccagt tagtacaaac ctataatcag ttgtttgaag agaaccctat aaatgcaagt
     1261 ggcgtggatg cgaaggctat tcttagcgcc cgcctctcta aatcccgacg gctagaaaac
     1321 ctgatcgcac aattacccgg agagaagaaa aatgggttgt tcggtaacct tatagcgctc
     1381 tcactaggcc tgacaccaaa ttttaagtcg aacttcgact tagctgaaga tgccaaattg
     1441 cagcttagta aggacacgta cgatgacgat ctcgacaatc tactggcaca aattggagat
     1501 cagtatgcgg acttattttt ggctgccaaa aaccttagcg atgcaatcct cctatctgac
     1561 atactgagag ttaatactga gattaccaag gcgccgttat ccgcttcaat gatcaaaagg
     1621 tacgatgaac atcaccaaga cttgacactt ctcaaggccc tagtccgtca gcaactgcct
     1681 gagaaatata aggaaatatt ctttgatcag tcgaaaaacg ggtacgcagg ttatattgac
     1741 ggcggagcga gtcaagagga attctacaag tttatcaaac ccatattaga gaagatggat
     1801 gggacggaag agttgcttgt aaaactcaat cgcgaagatc tactgcgaaa gcagcggact
     1861 ttcgacaacg gtagcattcc acatcaaatc cacttaggcg aattgcatgc tatacttaga
     1921 aggcaggagg atttttatcc gttcctcaaa gacaatcgtg aaaagattga gaaaatccta
     1981 acctttcgca taccttacta tgtgggaccc ctggcccgag ggaactctcg gttcgcatgg
     2041 atgacaagaa agtccgaaga aacgattact ccatggaatt ttgaggaagt tgtcgataaa
     2101 ggtgcgtcag ctcaatcgtt catcgagagg atgaccaact ttgacaagaa tttaccgaac
     2161 gaaaaagtat tgcctaagca cagtttactt tacgagtatt tcacagtgta caatgaactc
     2221 acgaaagtta agtatgtcac tgagggcatg cgtaaacccg cctttctaag cggagaacag
     2281 aagaaagcaa tagtagatct gttattcaag accaaccgca aagtgacagt taagcaattg
     2341 aaagaggact actttaagaa aattgaatgc ttcgattctg tcgagatctc cggggtagaa
     2401 gatcgattta atgcgtcact tggtacgtat catgacctcc taaagataat taaagataag
     2461 gacttcctgg ataacgaaga gaatgaagat atcttagaag atatagtgtt gactcttacc
     2521 ctctttgaag atcgggaaat gattgaggaa agactaaaaa catacgctca cctgttcgac
     2581 gataaggtta tgaaacagtt aaagaggcgt cgctatacgg gctggggacg attgtcgcgg
     2641 aaacttatca acgggataag agacaagcaa agtggtaaaa ctattctcga ttttctaaag
     2701 agcgacggct tcgccaatag gaactttatg cagctgatcc atgatgactc tttaaccttc
     2761 aaagaggata tacaaaaggc acaggtttcc ggacaagggg actcattgca cgaacatatt
     2821 gcgaatcttg ctggttcgcc agccatcaaa aagggcatac tccagacagt caaagtagtg
     2881 gatgagctag ttaaggtcat gggacgtcac aaaccggaaa acattgtaat cgagatggca
     2941 cgcgaaaatc aaacgactca gaaggggcaa aaaaacagtc gagagcggat gaagagaata
     3001 gaagagggta ttaaagaact gggcagccag atcttaaagg agcatcctgt ggaaaatacc
     3061 caattgcaga acgagaaact ttacctctat tacctacaaa atggaaggga catgtatgtt
     3121 gatcaggaac tggacataaa ccgtttatct gattacgacg tcgatcacat tgtaccccaa
     3181 tcctttttga aggacgattc aatcgacaat aaagtgctta cacgctcgga taagaaccga
     3241 gggaaaagtg acaatgttcc aagcgaggaa gtcgtaaaga aaatgaagaa ctattggcgg
     3301 cagctcctaa atgcgaaact gataacgcaa agaaagttcg ataacttaac taaagctgag
     3361 aggggtggct tgtctgaact tgacaaggcc ggatttatta aacgtcagct cgtggaaacc
     3421 cgccaaatca caaagcatgt tgcacagata ctagattccc gaatgaatac gaaatacgac
     3481 gagaacgata agctgattcg ggaagtcaaa gtaatcactt taaagtcaaa attggtgtcg
     3541 gacttcagaa aggattttca attctataaa gttagggaga taaataacta ccaccatgcg
     3601 cacgacgctt atcttaatgc cgtcgtaggg accgcactca ttaagaaata cccgaagcta
     3661 gaaagtgagt ttgtgtatgg tgattacaaa gtttatgacg tccgtaagat gatcgcgaaa
     3721 agcgaacagg agataggcaa ggctacagcc aaatacttct tttattctaa cattatgaat
     3781 ttctttaaga cggaaatcac tctggcaaac ggagagatac gcaaacgacc tttaattgaa
     3841 accaatgggg agacaggtga aatcgtatgg gataagggcc gggacttcgc gacggtgaga
     3901 aaagttttgt ccatgcccca agtcaacata gtaaagaaaa ctgaggtgca gaccggaggg
     3961 ttttcaaagg aatcgattct tccaaaaagg aatagtgata agctcatcgc tcgtaaaaag
     4021 gactgggacc cgaaaaagta cggtggcttc gatagcccta cagttgccta ttctgtccta
     4081 gtagtggcaa aagttgagaa gggaaaatcc aagaaactga agtcagtcaa agaattattg
     4141 gggataacga ttatggagcg ctcgtctttt gaaaagaacc ccatcgactt ccttgaggcg
     4201 aaaggttaca aggaagtaaa aaaggatctc ataattaaac taccaaagta tagtctgttt
     4261 gagttagaaa atggccgaaa acggatgttg gctagcgccg gagagcttca aaaggggaac
     4321 gaactcgcac taccgtctaa atacgtgaat ttcctgtatt tagcgtccca ttacgagaag
     4381 ttgaaaggtt cacctgaaga taacgaacag aagcaacttt ttgttgagca gcacaaacat
     4441 tatctcgacg aaatcataga gcaaatttcg gaattcagta agagagtcat cctagctgat
     4501 gccaatctgg acaaagtatt aagcgcatac aacaagcaca gggataaacc catacgtgag
     4561 caggcggaaa atattatcca tttgtttact cttaccaacc tcggcgctcc agccgcattc
     4621 aagtattttg acacaacgat agatcgcaaa cgatacactt ctaccaagga ggtgctagac
     4681 gcgacactga ttcaccaatc catcacggga ttatatgaaa ctcggataga tttgtcacag
     4741 cttgggggtg actctggtgg ttctcccaag aagaagagga aagtcgctag cggcagcggc
     4801 gccaccaact tcagcctgct gaagcaggcc ggcgacgtgg aggagaaccc cggccccatg
     4861 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     4921 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     4981 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     5041 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     5101 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     5161 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     5221 gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg tctgggcagc
     5281 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     5341 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     5401 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaacg
     5461 cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5521 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5581 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5641 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5701 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5761 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5821 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     5881 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5941 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     6001 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6061 cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     6121 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
     6181 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6241 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6301 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6361 ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6421 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     6481 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6541 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     6601 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6661 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6721 aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac
     6781 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     6841 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     6901 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     6961 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     7021 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     7081 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     7141 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     7201 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     7261 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     7321 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     7381 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     7441 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     7501 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     7561 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     7621 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     7681 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     7741 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     7801 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     7861 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7921 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7981 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     8041 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     8101 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     8161 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     8221 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     8281 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     8341 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     8401 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     8461 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     8521 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     8581 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     8641 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     8701 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     8761 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     8821 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8881 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8941 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     9001 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     9061 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     9121 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     9181 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     9241 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     9301 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     9361 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     9421 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     9481 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     9541 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
     9601 agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt
     9661 aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt
     9721 ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat
     9781 tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata
     9841 cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     9901 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     9961 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
    10021 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
    10081 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
    10141 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
    10201 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
    10261 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
    10321 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
    10381 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
    10441 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
    10501 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
    10561 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
    10621 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
    10681 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
    10741 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
    10801 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
    10861 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
    10921 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
    10981 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
    11041 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
    11101 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
    11161 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
    11221 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
    11281 atattcacca ttatcgtttc agacccacct cccaaccccg aggggaccca gaccacaatt
    11341 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
    11401 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
    11461 tttattacag ggacagcaga gatccacttt ggcgccggct cgagtggctc cggtgcccgt
    11521 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
    11581 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
    11641 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
    11701 gttctttttc gcaacgggtt tgccgccaga acacaggtgt cgtgacgcgg gatccgccac
    11761 catgtccgaa gtcgagtttt cccatgagta ctggatgaga cacgcattga ctctcgcaaa
    11821 gagggcttgg gatgaacgcg aggtgcccgt gggggcagta ctcgtgcata acaatcgcgt
    11881 aatcggcgaa ggttggaata ggccgatcgg acgccacgac cccactgcac atgcggaaat
    11941 catggccctt cgacagggag ggcttgtgat gcagaattat cgacttatcg atgcgacgct
    12001 gtacgtcacg cttgaacctt gcgtaatgtg cgcgggagct atgattcact cccgcattgg
    12061 acgagttgta ttcggtgccc gcgacgccaa gacgggtgcc gcaggttcac tgatggacgt
    12121 gctgcatcac ccaggcatga accaccgggt agaaatcaca gaaggcatat tggcggacga
    12181 atgtgcggcg ctgttgtccg acttttttcg catgcggagg caggagatca aggcccagaa
    12241 aaaagcacaa tcctctactg actctggtgg ttcttctggt ggttctagcg gcagcgagac
    12301 t
//
