LOCUS       sequence-314848-        9088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ABE7.10-P2A-GFP by EFS promoter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9088)
  AUTHORS   Shin HR, See JE, Kweon J, Kim HS, Sung GJ, Park S, Jang AH, Jang G, 
            Choi KC, Kim I, Kim JS, Kim Y
  TITLE     Small-molecule inhibitors of histone deacetylase improve 
            CRISPR-based adenine base editing.
  JOURNAL   Nucleic Acids Res. 2021 Feb 26;49(4):2390-2399. doi: 
            10.1093/nar/gkab052.
  PUBMED    33544854
REFERENCE   2  (bases 1 to 9088)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2021-02-26- 26"; volume: "49"; issue: 
            "4"; pages: "2390-2399"
COMMENT     Sequence Label: EFS-ABE7.10-P2A-GFP
FEATURES             Location/Qualifiers
     source          1..9088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1098..4496,4497..4499,4500..5096,5097,5098..5100,
                     5101..5102,5103..5105,5106..5198)
                     /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(1098..1121,1122..1124,1125..5198)
                     /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             5199..5255
                     /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(5256..5258,5259..5261,5262..5972)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5256..5258,5259..5261,5262..5972)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5301..5322)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5562..5581)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5909..5930
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5985..6005
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6014..6031
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6054..6071)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6060..6284
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6355..6371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6355..6371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6368..6390)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6379..6395
                     /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)."
     promoter        complement(join(6403..6409,6410..6427,6428..6433))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6448..6469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6586..6603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6757..7345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6837..6856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7516..8307,8308..8376))
                     /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     8139..8158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8377..8481)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8560..8579)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        8745..8956
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     8963..8982
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8963..8981
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(8995..9088,1..407,408..503,504..1001,1002..1097)
                     /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 cagtactcgt gcataacaat cgcgtaatcg gcgaaggttg gaataggccg atcggacgcc
       61 acgaccccac tgcacatgcg gaaatcatgg cccttcgaca gggagggctt gtgatgcaga
      121 attatcgact tatcgatgcg acgctgtacg tcacgcttga accttgcgta atgtgcgcgg
      181 gagctatgat tcactcccgc attggacgag ttgtattcgg tgcccgcgac gccaagacgg
      241 gtgccgcagg ttcactgatg gacgtgctgc atcacccagg catgaaccac cgggtagaaa
      301 tcacagaagg catattggcg gacgaatgtg cggcgctgtt gtccgacttt tttcgcatgc
      361 ggaggcagga gatcaaggcc cagaaaaaag cacaatcctc tactgactct ggtggttctt
      421 ctggtggttc tagcggcagc gagactcccg ggacctcaga gtccgccaca cccgaaagtt
      481 ctggtggttc ttctggtggt tcttccgaag tcgagttttc ccatgagtac tggatgagac
      541 acgcattgac tctcgcaaag agggctcgag atgaacgcga ggtgcccgtg ggggcagtac
      601 tcgtgctcaa caatcgcgta atcggcgaag gttggaatag ggcaatcgga ctccacgacc
      661 ccactgcaca tgcggaaatc atggcccttc gacagggagg gcttgtgatg cagaattatc
      721 gacttatcga tgcgacgctg tacgtcacgt ttgaaccttg cgtaatgtgc gcgggagcta
      781 tgattcactc ccgcattgga cgagttgtat tcggtgttcg caacgccaag acgggtgccg
      841 caggttcact gatggacgtg ctgcattacc caggcatgaa ccaccgggta gaaatcacag
      901 aaggcatatt ggcggacgaa tgtgcggcgc tgttgtgtta cttttttcgc atgcccaggc
      961 aggtctttaa cgcccagaaa aaagcacaat cctctactga ctctggtggt tcttctggtg
     1021 gttctagcgg cagcgagact cccgggacct cagagtccgc cacacccgaa agttctggtg
     1081 gttcttctgg tggttctgat aaaaagtatt ctattggttt agccatcggc actaattccg
     1141 ttggatgggc tgtcataacc gatgaataca aagtaccttc aaagaaattt aaggtgttgg
     1201 ggaacacaga ccgtcattcg attaaaaaga atcttatcgg tgccctccta ttcgatagtg
     1261 gcgaaacggc agaggcgact cgcctgaaac gaaccgctcg gagaaggtat acacgtcgca
     1321 agaaccgaat atgttactta caagaaattt ttagcaatga gatggccaaa gttgacgatt
     1381 ctttctttca ccgtttggaa gagtccttcc ttgtcgaaga ggacaagaaa catgaacggc
     1441 accccatctt tggaaacata gtagatgagg tggcatatca tgaaaagtac ccaacgattt
     1501 atcacctcag aaaaaagcta gttgactcaa ctgataaagc ggacctgagg ttaatctact
     1561 tggctcttgc ccatatgata aagttccgtg ggcactttct cattgagggt gatctaaatc
     1621 cggacaactc ggatgtcgac aaactgttca tccagttagt acaaacctat aatcagttgt
     1681 ttgaagagaa ccctataaat gcaagtggcg tggatgcgaa ggctattctt agcgcccgcc
     1741 tctctaaatc ccgacggcta gaaaacctga tcgcacaatt acccggagag aagaaaaatg
     1801 ggttgttcgg taaccttata gcgctctcac taggcctgac accaaatttt aagtcgaact
     1861 tcgacttagc tgaagatgcc aaattgcagc ttagtaagga cacgtacgat gacgatctcg
     1921 acaatctact ggcacaaatt ggagatcagt atgcggactt atttttggct gccaaaaacc
     1981 ttagcgatgc aatcctccta tctgacatac tgagagttaa tactgagatt accaaggcgc
     2041 cgttatccgc ttcaatgatc aaaaggtacg atgaacatca ccaagacttg acacttctca
     2101 aggccctagt ccgtcagcaa ctgcctgaga aatataagga aatattcttt gatcagtcga
     2161 aaaacgggta cgcaggttat attgacggcg gagcgagtca agaggaattc tacaagttta
     2221 tcaaacccat attagagaag atggatggga cggaagagtt gcttgtaaaa ctcaatcgcg
     2281 aagatctact gcgaaagcag cggactttcg acaacggtag cattccacat caaatccact
     2341 taggcgaatt gcatgctata cttagaaggc aggaggattt ttatccgttc ctcaaagaca
     2401 atcgtgaaaa gattgagaaa atcctaacct ttcgcatacc ttactatgtg ggacccctgg
     2461 cccgagggaa ctctcggttc gcatggatga caagaaagtc cgaagaaacg attactccat
     2521 ggaattttga ggaagttgtc gataaaggtg cgtcagctca atcgttcatc gagaggatga
     2581 ccaactttga caagaattta ccgaacgaaa aagtattgcc taagcacagt ttactttacg
     2641 agtatttcac agtgtacaat gaactcacga aagttaagta tgtcactgag ggcatgcgta
     2701 aacccgcctt tctaagcgga gaacagaaga aagcaatagt agatctgtta ttcaagacca
     2761 accgcaaagt gacagttaag caattgaaag aggactactt taagaaaatt gaatgcttcg
     2821 attctgtcga gatctccggg gtagaagatc gatttaatgc gtcacttggt acgtatcatg
     2881 acctcctaaa gataattaaa gataaggact tcctggataa cgaagagaat gaagatatct
     2941 tagaagatat agtgttgact cttaccctct ttgaagatcg ggaaatgatt gaggaaagac
     3001 taaaaacata cgctcacctg ttcgacgata aggttatgaa acagttaaag aggcgtcgct
     3061 atacgggctg gggacgattg tcgcggaaac ttatcaacgg gataagagac aagcaaagtg
     3121 gtaaaactat tctcgatttt ctaaagagcg acggcttcgc caataggaac tttatgcagc
     3181 tgatccatga tgactcttta accttcaaag aggatataca aaaggcacag gtttccggac
     3241 aaggggactc attgcacgaa catattgcga atcttgctgg ttcgccagcc atcaaaaagg
     3301 gcatactcca gacagtcaaa gtagtggatg agctagttaa ggtcatggga cgtcacaaac
     3361 cggaaaacat tgtaatcgag atggcacgcg aaaatcaaac gactcagaag gggcaaaaaa
     3421 acagtcgaga gcggatgaag agaatagaag agggtattaa agaactgggc agccagatct
     3481 taaaggagca tcctgtggaa aatacccaat tgcagaacga gaaactttac ctctattacc
     3541 tacaaaatgg aagggacatg tatgttgatc aggaactgga cataaaccgt ttatctgatt
     3601 acgacgtcga tcacattgta ccccaatcct ttttgaagga cgattcaatc gacaataaag
     3661 tgcttacacg ctcggataag aaccgaggga aaagtgacaa tgttccaagc gaggaagtcg
     3721 taaagaaaat gaagaactat tggcggcagc tcctaaatgc gaaactgata acgcaaagaa
     3781 agttcgataa cttaactaaa gctgagaggg gtggcttgtc tgaacttgac aaggccggat
     3841 ttattaaacg tcagctcgtg gaaacccgcc aaatcacaaa gcatgttgca cagatactag
     3901 attcccgaat gaatacgaaa tacgacgaga acgataagct gattcgggaa gtcaaagtaa
     3961 tcactttaaa gtcaaaattg gtgtcggact tcagaaagga ttttcaattc tataaagtta
     4021 gggagataaa taactaccac catgcgcacg acgcttatct taatgccgtc gtagggaccg
     4081 cactcattaa gaaatacccg aagctagaaa gtgagtttgt gtatggtgat tacaaagttt
     4141 atgacgtccg taagatgatc gcgaaaagcg aacaggagat aggcaaggct acagccaaat
     4201 acttctttta ttctaacatt atgaatttct ttaagacgga aatcactctg gcaaacggag
     4261 agatacgcaa acgaccttta attgaaacca atggggagac aggtgaaatc gtatgggata
     4321 agggccggga cttcgcgacg gtgagaaaag ttttgtccat gccccaagtc aacatagtaa
     4381 agaaaactga ggtgcagacc ggagggtttt caaaggaatc gattcttcca aaaaggaata
     4441 gtgataagct catcgctcgt aaaaaggact gggacccgaa aaagtacggt ggcttcgata
     4501 gccctacagt tgcctattct gtcctagtag tggcaaaagt tgagaaggga aaatccaaga
     4561 aactgaagtc agtcaaagaa ttattgggga taacgattat ggagcgctcg tcttttgaaa
     4621 agaaccccat cgacttcctt gaggcgaaag gttacaagga agtaaaaaag gatctcataa
     4681 ttaaactacc aaagtatagt ctgtttgagt tagaaaatgg ccgaaaacgg atgttggcta
     4741 gcgccggaga gcttcaaaag gggaacgaac tcgcactacc gtctaaatac gtgaatttcc
     4801 tgtatttagc gtcccattac gagaagttga aaggttcacc tgaagataac gaacagaagc
     4861 aactttttgt tgagcagcac aaacattatc tcgacgaaat catagagcaa atttcggaat
     4921 tcagtaagag agtcatccta gctgatgcca atctggacaa agtattaagc gcatacaaca
     4981 agcacaggga taaacccata cgtgagcagg cggaaaatat tatccatttg tttactctta
     5041 ccaacctcgg cgctccagcc gcattcaagt attttgacac aacgatagat cgcaaacgat
     5101 acacttctac caaggaggtg ctagacgcga cactgattca ccaatccatc acgggattat
     5161 atgaaactcg gatagatttg tcacagcttg ggggtgacgc tactaacttc agcctgctga
     5221 agcaggctgg agacgtggag gagaaccctg gacctatggt gagcaagggc gaggagctgt
     5281 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     5341 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     5401 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctatggag
     5461 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     5521 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     5581 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     5641 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     5701 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     5761 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     5821 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     5881 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     5941 ggatcactct cggcatggac gagctgtaca agtctggtgg ttctcccaag aagaagagga
     6001 aagtctaacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     6061 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     6121 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     6181 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     6241 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     6301 aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6361 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6421 cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6481 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6541 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6601 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6661 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7381 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7441 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7501 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7561 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7621 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7681 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7741 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7801 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7861 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7921 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7981 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8041 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8101 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8161 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8221 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8281 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8341 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8401 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8461 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     8521 gggagatcga tctcccgatc ccctagggtc gactctcagt acaatctgct ctgatgccgc
     8581 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     8641 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     8701 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatagggcag agcgcacatc
     8761 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgatccggtg cctagagaag
     8821 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     8881 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     8941 tgccgccaga acacagggcc gctaatacga ctcactatag ggagagccgc caccatgtcc
     9001 gaagtcgagt tttcccatga gtactggatg agacacgcat tgactctcgc aaagagggct
     9061 tgggatgaac gcgaggtgcc cgtggggg
//
