LOCUS       sequence-451281-       10302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ABEmax plasmid enabling A:T to G:C base editing using a single 
            plasmid. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10302)
  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 10302)
  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 V2
FEATURES             Location/Qualifiers
     source          1..10302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(114..137,138..140,141..4214)
                     /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             114..4214
                     /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             4215..4262
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4278..4331
                     /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             4332..4931
                     /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(4332..4351)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4828..4848
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4959..4976)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4965..5172
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5181..5321
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5181..5310
                     /label=AAV2 ITR
     rep_origin      5396..5851
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5483..5502)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5693..5714
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5868..5887)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5987..6009
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6047..6065)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6133..6237
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6238..6306,6307..7098)
                     /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(6456..6475)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7269..7857
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7758..7777
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7919..8167
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7919..7939
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8090..8109
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8186..8261
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8358..8643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        8645..8922
                     /label=chicken beta-actin promoter
     intron          8923..9150
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      9162..9171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9207..9227
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cccagagctc caccgactcc ggaggatcta gcggaggctc ctctggctct gagacacctg
       61 gcacaagcga gagcgcaaca cctgaaagca gcgggggcag cagcgggggg tcagacaaga
      121 agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg
      181 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
      241 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
      301 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
      361 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
      421 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
      481 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
      541 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
      601 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
      661 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
      721 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
      781 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc
      841 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
      901 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
      961 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     1021 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     1081 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     1141 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
     1201 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     1261 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     1321 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     1381 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     1441 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     1501 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     1561 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     1621 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
     1681 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     1741 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     1801 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     1861 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     1921 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     1981 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     2041 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     2101 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     2161 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     2221 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     2281 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     2341 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     2401 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     2461 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     2521 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     2581 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
     2641 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
     2701 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     2761 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
     2821 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     2881 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     2941 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     3001 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     3061 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     3121 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     3181 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     3241 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     3301 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
     3361 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     3421 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
     3481 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     3541 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     3601 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     3661 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     3721 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
     3781 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     3841 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
     3901 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     3961 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
     4021 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     4081 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
     4141 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     4201 agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa
     4261 aggaattcgg cagtggagag ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga
     4321 atcctggccc aatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4381 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4441 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4501 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     4561 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     4621 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     4681 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac caccagggca
     4741 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     4801 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     4861 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     4921 ccggtgcctg agaattctaa ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     4981 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     5041 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     5101 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag
     5161 gcatgctggg gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     5221 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     5281 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     5341 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     5401 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5461 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5521 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5581 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     5641 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5701 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     5761 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5821 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     5881 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5941 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     6001 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     6061 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6121 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6181 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6241 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6301 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6361 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6421 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6481 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6541 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6601 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6661 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6721 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6781 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6841 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6901 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc
     6961 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7021 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7081 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7141 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7201 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7261 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7321 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7381 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7441 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7501 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7561 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7621 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7681 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7741 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7801 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7861 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtga
     7921 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     7981 aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     8041 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     8101 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     8161 aaacaccggg tcttcgagaa gacctgtttt agagctagaa atagcaagtt aaaataaggc
     8221 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt ttagagctag
     8281 aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt ctgcagacaa
     8341 atggctctag aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     8401 aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa
     8461 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     8521 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac
     8581 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     8641 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
     8701 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
     8761 ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga
     8821 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
     8881 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
     8941 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
     9001 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg
     9061 agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg
     9121 gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg gacaaacgga
     9181 cagccgacgg aagcgagttc gagtcaccaa agaagaagcg gaaagtctct gaagtcgagt
     9241 ttagccacga gtattggatg aggcacgcac tgaccctggc aaagcgagca tgggatgaaa
     9301 gagaagtccc cgtgggcgcc gtgctggtgc acaacaatag agtgatcgga gagggatgga
     9361 acaggccaat cggccgccac gaccctaccg cacacgcaga gatcatggca ctgaggcagg
     9421 gaggcctggt catgcagaat taccgcctga tcgatgccac cctgtatgtg acactggagc
     9481 catgcgtgat gtgcgcagga gcaatgatcc acagcaggat cggaagagtg gtgttcggag
     9541 cacgggacgc caagaccggc gcagcaggct ccctgatgga tgtgctgcac caccccggca
     9601 tgaaccaccg ggtggagatc acagagggaa tcctggcaga cgagtgcgcc gccctgctga
     9661 gcgatttctt tagaatgcgg agacaggaga tcaaggccca gaagaaggca cagagctcca
     9721 ccgactctgg aggatctagc ggaggatcct ctggaagcga gacaccaggc acaagcgagt
     9781 ccgccacacc agagagctcc ggcggctcct ccggaggatc ctctgaggtg gagttttccc
     9841 acgagtactg gatgagacat gccctgaccc tggccaagag ggcacgcgat gagagggagg
     9901 tgcctgtggg agccgtgctg gtgctgaaca atagagtgat cggcgagggc tggaacagag
     9961 ccatcggcct gcacgaccca acagcccatg ccgaaattat ggccctgaga cagggcggcc
    10021 tggtcatgca gaactacaga ctgattgacg ccaccctgta cgtgacattc gagccttgcg
    10081 tgatgtgcgc cggcgccatg atccactcta ggatcggccg cgtggtgttt ggcgtgagga
    10141 acgcaaaaac cggcgccgca ggctccctga tggacgtgct gcactacccc ggcatgaatc
    10201 accgcgtcga aattaccgag ggaatcctgg cagatgaatg tgccgccctg ctgtgctatt
    10261 tctttcggat gcctagacag gtgttcaatg ctcagaagaa gg
//
