LOCUS       sequence-464531-       10301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ABEmax plasmid enabling A:T to G:C base editing using a single 
            plasmid. sgRNA uses 3TC scaffold for increased sgRNA expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10301)
  AUTHORS   Chey YCJ, Gierus L, Lushington C, Arudkumar JC, B Geiger A, Staker 
            LG, Robertson LJ, Pfitzner C, Kennedy JG, Lee RHB, Godahewa GI, 
            Adikusuma F, Thomas PQ
  TITLE     Optimal SpCas9- and SaCas9-mediated gene editing by enhancing gRNA 
            transcript levels through scaffold poly-T tract reduction.
  JOURNAL   BMC Genomics. 2025 Feb 12;26(1):138. doi: 
            10.1186/s12864-025-11317-2.
  PUBMED    39939860
REFERENCE   2  (bases 1 to 10301)
  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: "BMC 
            Genomics."; date: "2025-02-12"; volume: "26(1):138. doi"; pages: " 
            10.1186/s12864-025-11317-2"
COMMENT     Sequence Label: ABEmax-Puro V3
FEATURES             Location/Qualifiers
     source          1..10301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             34..54
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1243..1266,1267..1269,1270..5343)
                     /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             1243..5343
                     /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             5344..5391
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5407..5460
                     /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             5461..6060
                     /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(5461..5480)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5957..5977
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6088..6105)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6094..6301
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6310..6450
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6310..6439
                     /label=AAV2 ITR
     rep_origin      6525..6980
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6612..6631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6822..6843
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6997..7016)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7116..7138
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7176..7194)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7262..7366
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7367..7435,7436..8227)
                     /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(7585..7604)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8398..8986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8887..8906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        9048..9296
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9048..9068
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9219..9238
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9315..9390
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        9487..9772
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        9774..10050
                     /label=chicken beta-actin promoter
     intron          10051..10278
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      10290..10299
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtctctgaa
       61 gtcgagttta gccacgagta ttggatgagg cacgcactga ccctggcaaa gcgagcatgg
      121 gatgaaagag aagtccccgt gggcgccgtg ctggtgcaca acaatagagt gatcggagag
      181 ggatggaaca ggccaatcgg ccgccacgac cctaccgcac acgcagagat catggcactg
      241 aggcagggag gcctggtcat gcagaattac cgcctgatcg atgccaccct gtatgtgaca
      301 ctggagccat gcgtgatgtg cgcaggagca atgatccaca gcaggatcgg aagagtggtg
      361 ttcggagcac gggacgccaa gaccggcgca gcaggctccc tgatggatgt gctgcaccac
      421 cccggcatga accaccgggt ggagatcaca gagggaatcc tggcagacga gtgcgccgcc
      481 ctgctgagcg atttctttag aatgcggaga caggagatca aggcccagaa gaaggcacag
      541 agctccaccg actctggagg atctagcgga ggatcctctg gaagcgagac accaggcaca
      601 agcgagtccg ccacaccaga gagctccggc ggctcctccg gaggatcctc tgaggtggag
      661 ttttcccacg agtactggat gagacatgcc ctgaccctgg ccaagagggc acgcgatgag
      721 agggaggtgc ctgtgggagc cgtgctggtg ctgaacaata gagtgatcgg cgagggctgg
      781 aacagagcca tcggcctgca cgacccaaca gcccatgccg aaattatggc cctgagacag
      841 ggcggcctgg tcatgcagaa ctacagactg attgacgcca ccctgtacgt gacattcgag
      901 ccttgcgtga tgtgcgccgg cgccatgatc cactctagga tcggccgcgt ggtgtttggc
      961 gtgaggaacg caaaaaccgg cgccgcaggc tccctgatgg acgtgctgca ctaccccggc
     1021 atgaatcacc gcgtcgaaat taccgaggga atcctggcag atgaatgtgc cgccctgctg
     1081 tgctatttct ttcggatgcc tagacaggtg ttcaatgctc agaagaaggc ccagagctcc
     1141 accgactccg gaggatctag cggaggctcc tctggctctg agacacctgg cacaagcgag
     1201 agcgcaacac ctgaaagcag cgggggcagc agcggggggt cagacaagaa gtacagcatc
     1261 ggcctggcca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
     1321 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
     1381 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
     1441 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
     1501 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     1561 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
     1621 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     1681 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     1741 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     1801 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
     1861 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     1921 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
     1981 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     2041 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     2101 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     2161 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     2221 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     2281 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
     2341 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     2401 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     2461 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     2521 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     2581 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     2641 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
     2701 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
     2761 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
     2821 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
     2881 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     2941 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
     3001 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
     3061 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
     3121 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
     3181 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
     3241 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
     3301 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
     3361 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
     3421 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
     3481 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     3541 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     3601 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     3661 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
     3721 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg
     3781 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
     3841 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
     3901 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
     3961 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
     4021 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
     4081 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
     4141 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
     4201 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
     4261 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
     4321 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
     4381 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
     4441 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
     4501 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
     4561 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
     4621 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
     4681 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
     4741 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
     4801 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
     4861 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
     4921 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
     4981 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
     5041 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
     5101 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
     5161 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
     5221 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
     5281 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
     5341 gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa ggaattcggc
     5401 agtggagagg gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca
     5461 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     5521 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     5581 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     5641 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5701 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     5761 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     5821 cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc accagggcaa gggtctgggc
     5881 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     5941 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     6001 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     6061 gaattctaac tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     6121 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     6181 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     6241 gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg
     6301 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     6361 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     6421 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     6481 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     6541 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6601 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6661 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6721 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6781 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6841 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     6901 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6961 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     7021 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     7081 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     7141 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     7201 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     7261 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     7321 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7381 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     7441 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7501 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7561 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     7621 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     7681 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     7741 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7801 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7861 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7921 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7981 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     8041 tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc
     8101 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     8161 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     8221 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     8281 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8341 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     8401 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     8461 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     8521 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     8581 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     8641 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     8701 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     8761 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     8821 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     8881 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     8941 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     9001 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtgag ggcctatttc
     9061 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attggaatta
     9121 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
     9181 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
     9241 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgggt
     9301 cttcgagaag acctgtttca gagctagaaa tagcaagttg aaataaggct agtccgttat
     9361 caacttgaaa aagtggcacc gagtcggtgc ttttttgttt tagagctaga aatagcaagt
     9421 taaaataagg ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa tggctctaga
     9481 ggtacccgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     9541 cccattgacg tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     9601 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     9661 tattgacgtc aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg
     9721 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     9781 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     9841 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg ggggcgcgcg
     9901 ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca
     9961 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
    10021 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
    10081 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
    10141 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag caagaggtaa
    10201 gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga gcacctgcct
    10261 gaaatcactt tttttcaggt tggaccggtg ccaccatgga c
//
