LOCUS       sequence-500925-       13898 bp    DNA     circular SYN 24-FEB-2026
DEFINITION  Expression of ABE-ARG8 fusion protein in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13898)
  AUTHORS   Yin S, Jarosz DF, Ting AY
  TITLE     Towards CRISPR-based editing of the mitochondrial genome in yeast.
  JOURNAL   Proc Natl Acad Sci U S A. 2026 Jan 20;123(3):e2505894123. doi: 
            10.1073/pnas.2505894123. Epub 2026 Jan 16.
  PUBMED    41543903
REFERENCE   2  (bases 1 to 13898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 24, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2026-01-20"; volume: "123(3):e2505894123. 
            doi"; pages: " 10.1073/pnas.2505894123. Epub 2026 Jan 16"
COMMENT     Sequence Label: MTS-ABE-ARG8
FEATURES             Location/Qualifiers
     source          1..13898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1055..1698
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     1592..1615
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     1704..1720
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1704..1720
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    1728..1752
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(2571..2594,2595..2597,2598..6671)
                     /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             2571..6671
                     /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             7953..7994
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7998..8054
                     /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             join(8055..8057,8058..8060,8061..8774)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKAEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8361..8380)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8708..8729
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      8781..9028
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(8797..8815)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(9046..9065)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9047..9065)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9075..9092)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9075..9091)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9084..9106)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9232..9687
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9319..9338)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9529..9550
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9988..10395
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             10396..11490
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(11984..12003)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12103..12125
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(12163..12181)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    12223..12726
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        12763..12867
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12868..12936,12937..13728)
                     /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(13086..13105)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctcag tttatcatta tcaatactcg ccatttcaaa
     1081 gaatacgtaa ataattaata gtagtgattt tcctaacttt atttagtcaa aaaattagcc
     1141 ttttaattct gctgtaaccc gtacatgccc aaaatagggg gcgggttaca cagaatatat
     1201 aacatcgtag gtgtctgggt gaacagttta ttcctggcat ccactaaata taatggagcc
     1261 cgctttttaa gctggcatcc agaaaaaaaa agaatcccag caccaaaata ttgttttctt
     1321 caccaaccat cagttcatag gtccattctc ttagcgcaac tacagagaac aggggcacaa
     1381 acaggcaaaa aacgggcaca acctcaatgg agtgatgcaa cctgcctgga gtaaatgatg
     1441 acacaaggca attgacccac gcatgtatct atctcatttt cttacacctt ctattacctt
     1501 ctgctctctc tgatttggaa aaagctgaaa aaaaaggttg aaaccagttc cctgaaatta
     1561 ttcccctact tgactaataa gtatataaag acggtaggta ttgattgtaa ttctgtaaat
     1621 ctatttctta aacttcttaa attctacttt tatagttagt ctttttttta gttttaaaac
     1681 accagaactt agtttcgacg gattctagaa ctagtggatc ccccatcaca agtttgtaca
     1741 aaaaagcagg cttcgaagga gataacaaaa tggcctccac tcgtgtcctc gcctctcgcc
     1801 tggcctccca gatggctgct tccgccaagg ttgcccgccc tgctgtccgc gttgctcagg
     1861 tcagcaagcg caccatccag actggctccc ccctccagac cctcaagcgc acccagatga
     1921 cctccatcgt caacgccacc acccgccagg ctttccagaa gcgcgcctac tcttcctctg
     1981 aggtggagtt ttcccacgag tactggatga gacatgccct gaccctggcc aagagggcac
     2041 gggatgagag ggaggtgcct gtgggagccg tgctggtgct gaacaataga gtgatcggcg
     2101 agggctggaa cagagccatc ggcctgcacg acccaacagc ccatgccgaa attatggccc
     2161 tgagacaggg cggcctggtc atgcagaact acagactgat tgacgccacc ctgtacgtga
     2221 cattcgagcc ttgcgtgatg tgcgccggcg ccatgatcca ctctaggatc ggccgcgtgg
     2281 tgtttggctg gaggaactca aaaagaggcg ccgcaggctc cctgatgaac gtgctgaact
     2341 accccggcat gaatcaccgc gtcgaaatta ccgagggaat cctggcagat gaatgtgccg
     2401 ccctgctgtg cgatttctat cggatgccta gacaggtgtt caatgctcag aagaaggccc
     2461 agagctccat caactccgga ggatctagcg gaggctcctc tggctctgag acacctggca
     2521 caagcgagag cgcaacacct gaaagcagcg ggggcagcag cggggggtca gacaagaagt
     2581 acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
     2641 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
     2701 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
     2761 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
     2821 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     2881 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     2941 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     3001 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     3061 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     3121 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     3181 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     3241 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga
     3301 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     3361 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     3421 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     3481 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     3541 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     3601 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     3661 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     3721 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
     3781 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
     3841 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
     3901 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
     3961 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
     4021 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
     4081 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
     4141 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
     4201 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
     4261 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
     4321 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
     4381 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
     4441 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
     4501 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
     4561 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
     4621 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
     4681 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
     4741 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
     4801 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
     4861 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
     4921 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc
     4981 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
     5041 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga
     5101 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg
     5161 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
     5221 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
     5281 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
     5341 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
     5401 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
     5461 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
     5521 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
     5581 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
     5641 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
     5701 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga
     5761 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
     5821 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
     5881 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
     5941 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg
     6001 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg
     6061 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
     6121 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
     6181 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg
     6241 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
     6301 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
     6361 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
     6421 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
     6481 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
     6541 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
     6601 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
     6661 tgggaggtga ctctggcggc tcatttaaaa gatatttagg gtccagcacg tcatcaagaa
     6721 gatttacaag cattttagag gaaaaggcct ttcaagtgac cacttactct agacctgaag
     6781 atctatgtat aactagaggt aaaaatgcaa agctgtatga tgacgtgaat ggtaaagaat
     6841 atatcgattt caccgcaggt attgcggtga ccgcattagg ccatgcaaat cctaaagtgg
     6901 cagaaattct gcaccatcag gctaacaaac tggttcattc ctccaacctt tacttcacta
     6961 aggaatgttt ggatttaagt gaaaagattg ttgaaaagac caagcaattc ggtggtcaac
     7021 acgacgcctc aagagtattt ttatgtaatt ctggtacgga agcaaatgaa gctgctttga
     7081 agtttgcaaa gaaacatggt ataatgaaaa atcctagcaa gcaaggcatt gttgcatttg
     7141 agaactcttt tcatggccgt actatgggcg ctttatctgt cacttggaat agtaaatata
     7201 gaactccttt tggggatttg gttccccatg tctcattctt aaatttgaat gacgaaatga
     7261 ccaaactaca aagttatatc gagaccaaaa aggacgagat tgctggttta attgtcgagc
     7321 ccatacaagg tgaaggtggg gtttttcccg tagaagttga aaagctaacc ggattgaaga
     7381 aaatatgtca agataatgat gtgattgtca ttcatgatga aattcaatgc ggtttgggcc
     7441 gttcaggtaa actatgggct catgcttatt taccaagtga ggctcatccg gatattttta
     7501 catctgccaa agcattggga aatggcttcc ccatcgctgc caccatcgtc aatgaaaaag
     7561 ttaataatgc tttgagagtt ggtgaccacg gcaccacgta tggtggtaat ccgctggcct
     7621 gttctgtaag caactatgtt ttggatacca tagcagacga agcttttttg aaacaagtct
     7681 ctaagaagag tgatatctta caaaagcgct tgcgcgaaat tcaagccaaa tatccaaatc
     7741 aaataaagac tatcagagga aaaggtttga tgcttggtgc tgagttcgtc gaaccaccca
     7801 ccgaggtcat caaaaaggcc agagaattgg gacttttgat cattaccgct ggtaagagta
     7861 ccgttagatt tgttcccgca ttaacgattg aagacgaact aatcgaagaa gggatggatg
     7921 cttttgaaaa ggctattgaa gcggtttacg ctggcaagcc catccccaac cccctgctgg
     7981 gcctggacag caccggtgca acaaacttct ccctcttgaa acaagcagga gatgttgagg
     8041 aaaatcccgg gcctatggtg agcaaggccg aggagctgtt caccggggtg gtgcccatcc
     8101 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     8161 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     8221 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     8281 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     8341 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     8401 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     8461 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     8521 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     8581 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     8641 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     8701 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     8761 agctgtacaa gtaactcgag tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     8821 ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     8881 ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt
     8941 ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     9001 tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta tagtgagtcg
     9061 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     9121 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     9181 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     9241 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     9301 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     9361 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     9421 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     9481 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     9541 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     9601 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     9661 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     9721 cggtatttca caccgcatat cgaccggtcg aggagaactt ctagtatatc tacataccta
     9781 atattattgc cttattaaaa atggaatccc aacaattaca tcaaaatcca cattctcttc
     9841 aaaatcaatt gtcctgtact tccttgttca tgtgtgttca aaaacgttat atttatagga
     9901 taattatact ctatttctca acaagtaatt ggttgtttgg ccgagcggtc taaggcgcct
     9961 gattcaagaa atatcttgac cgcagttaac tgtgggaata ctcaggtatc gtaagatgca
    10021 agagttcgaa tctcttagca accattattt ttttcctcaa cataacgaga acacacaggg
    10081 gcgctatcgc acagaatcaa attcgatgac tggaaatttt ttgttaattt cagaggtcgc
    10141 ctgacgcata tacctttttc aactgaaaaa ttgggagaaa aaggaaaggt gagagcgccg
    10201 gaaccggctt ttcatataga atagagaagc gttcatgact aaatgcttgc atcacaatac
    10261 ttgaagttga caatattatt taaggaccta ttgttttttc caataggtgg ttagcaatcg
    10321 tcttactttc taacttttct taccttttac atttcagcaa tatatatata tatatttcaa
    10381 ggatatacca ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg tgaccacgtt
    10441 ggtcaagaaa tcacagccga agccattaag gttcttaaag ctatttctga tgttcgttcc
    10501 aatgtcaagt tcgatttcga aaatcattta attggtggtg ctgctatcga tgctacaggt
    10561 gttccacttc cagatgaggc gctggaagcc tccaagaagg ctgatgccgt tttgttaggt
    10621 gctgtgggtg gtcctaaatg gggtaccggt agtgttagac ctgaacaagg tttactaaaa
    10681 atccgtaaag aacttcaatt gtacgccaac ttaagaccat gtaactttgc atccgactct
    10741 cttttagact tatctccaat caagccacaa tttgctaaag gtactgactt cgttgttgtc
    10801 agagaattag tgggaggtat ttactttggt aagagaaagg aagacgatgg tgatggtgtc
    10861 gcttgggata gtgaacaata caccgttcca gaagtgcaaa gaatcacaag aatggccgct
    10921 ttcatggccc tacaacatga gccaccattg cctatttggt ccttggataa agctaatgtt
    10981 ttggcctctt caagattatg gagaaaaact gtggaggaaa ccatcaagaa cgaattccct
    11041 acattgaagg ttcaacatca attgattgat tctgccgcca tgatcctagt taagaaccca
    11101 acccacctaa atggtattat aatcaccagc aacatgtttg gtgatatcat ctccgatgaa
    11161 gcctccgtta tcccaggttc cttgggtttg ttgccatctg cgtccttggc ctctttgcca
    11221 gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg gttctgctcc agatttgcca
    11281 aagaataagg tcaaccctat cgccactatc ttgtctgctg caatgatgtt gaaattgtca
    11341 ttgaacttgc ctgaagaagg taaggccatt gaagatgcag ttaaaaaggt tttggatgca
    11401 ggtatcagaa ctggtgattt aggtggttcc aacagtacca ccgaagtcgg tgatgctgtc
    11461 gccgaagaag ttaagaaaat ccttgcttaa aaagattctc tttttttatg atatttgtac
    11521 ataaacttta taaatgaaat tcataataga aacgacacga aattacaaaa tggaatatgt
    11581 tcatagggta gacgaaacta tatacgcaat ctacatacat ttatcaagaa ggagaaaaag
    11641 gaggatgtaa aggaatacag gtaagcaaat tgatactaat ggctcaacgt gataaggaaa
    11701 aagaattgca ctttaacatt aatattgaca aggaggaggg caccacacaa aaagttaggt
    11761 gtaacagaaa atcatgaaac tatgattcct aatttatata ttggaggatt ttctctaaaa
    11821 aaaaaaaaat acaacaaata aaaaacactc aatgacctga ccatttgatg gagtttaagt
    11881 caataccttc ttgaaccatt tcccataatg gtgaaagttc cctcaagaat tttactctgt
    11941 cagaaacggc cttaacgacg tagtcgatat ggtgcactct cagtacaatc tgctctgatg
    12001 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
    12061 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
    12121 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
    12181 ttttataggt taatgtcatg ataataatgg tttcttagga cggatcgctt gcctgtaact
    12241 tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt
    12301 atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga
    12361 atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata
    12421 tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa
    12481 aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg
    12541 gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc
    12601 tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact
    12661 ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca
    12721 cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
    12781 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
    12841 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
    12901 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
    12961 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
    13021 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
    13081 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
    13141 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
    13201 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
    13261 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
    13321 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
    13381 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
    13441 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
    13501 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
    13561 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
    13621 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
    13681 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
    13741 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
    13801 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
    13861 tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
