LOCUS       sequence-319259-        9501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ABE7.10-P2A-GFP by CMV promoter in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9501)
  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 9501)
  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: pCMV-ABE7.10-P2A-GFP
FEATURES             Location/Qualifiers
     source          1..9501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(658..1158,1159..1254,1255..1752,1753..1848)
                     /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"
     CDS             join(1849..5247,5248..5250,5251..5847,5848,5849..5851,
                     5852..5853,5854..5856,5857..5949)
                     /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(1849..1872,1873..1875,1876..5949)
                     /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             5950..6006
                     /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(6007..6009,6010..6012,6013..6723)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6007..6009,6010..6012,6013..6723)
                     /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(6052..6073)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6313..6332)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6660..6681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6736..6756
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6765..6782
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6805..6822)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6811..7035
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7106..7122)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7106..7122)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7119..7141)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7130..7146
                     /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(7154..7160,7161..7178,7179..7184))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7199..7220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7337..7354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7508..8096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7588..7607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8267..9058,9059..9127))
                     /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     8890..8909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9128..9232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9311..9330)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 tccgaagtcg agttttccca tgagtactgg atgagacacg cattgactct cgcaaagagg
      721 gcttgggatg aacgcgaggt gcccgtgggg gcagtactcg tgcataacaa tcgcgtaatc
      781 ggcgaaggtt ggaataggcc gatcggacgc cacgacccca ctgcacatgc ggaaatcatg
      841 gcccttcgac agggagggct tgtgatgcag aattatcgac ttatcgatgc gacgctgtac
      901 gtcacgcttg aaccttgcgt aatgtgcgcg ggagctatga ttcactcccg cattggacga
      961 gttgtattcg gtgcccgcga cgccaagacg ggtgccgcag gttcactgat ggacgtgctg
     1021 catcacccag gcatgaacca ccgggtagaa atcacagaag gcatattggc ggacgaatgt
     1081 gcggcgctgt tgtccgactt ttttcgcatg cggaggcagg agatcaaggc ccagaaaaaa
     1141 gcacaatcct ctactgactc tggtggttct tctggtggtt ctagcggcag cgagactccc
     1201 gggacctcag agtccgccac acccgaaagt tctggtggtt cttctggtgg ttcttccgaa
     1261 gtcgagtttt cccatgagta ctggatgaga cacgcattga ctctcgcaaa gagggctcga
     1321 gatgaacgcg aggtgcccgt gggggcagta ctcgtgctca acaatcgcgt aatcggcgaa
     1381 ggttggaata gggcaatcgg actccacgac cccactgcac atgcggaaat catggccctt
     1441 cgacagggag ggcttgtgat gcagaattat cgacttatcg atgcgacgct gtacgtcacg
     1501 tttgaacctt gcgtaatgtg cgcgggagct atgattcact cccgcattgg acgagttgta
     1561 ttcggtgttc gcaacgccaa gacgggtgcc gcaggttcac tgatggacgt gctgcattac
     1621 ccaggcatga accaccgggt agaaatcaca gaaggcatat tggcggacga atgtgcggcg
     1681 ctgttgtgtt acttttttcg catgcccagg caggtcttta acgcccagaa aaaagcacaa
     1741 tcctctactg actctggtgg ttcttctggt ggttctagcg gcagcgagac tcccgggacc
     1801 tcagagtccg ccacacccga aagttctggt ggttcttctg gtggttctga taaaaagtat
     1861 tctattggtt tagccatcgg cactaattcc gttggatggg ctgtcataac cgatgaatac
     1921 aaagtacctt caaagaaatt taaggtgttg gggaacacag accgtcattc gattaaaaag
     1981 aatcttatcg gtgccctcct attcgatagt ggcgaaacgg cagaggcgac tcgcctgaaa
     2041 cgaaccgctc ggagaaggta tacacgtcgc aagaaccgaa tatgttactt acaagaaatt
     2101 tttagcaatg agatggccaa agttgacgat tctttctttc accgtttgga agagtccttc
     2161 cttgtcgaag aggacaagaa acatgaacgg caccccatct ttggaaacat agtagatgag
     2221 gtggcatatc atgaaaagta cccaacgatt tatcacctca gaaaaaagct agttgactca
     2281 actgataaag cggacctgag gttaatctac ttggctcttg cccatatgat aaagttccgt
     2341 gggcactttc tcattgaggg tgatctaaat ccggacaact cggatgtcga caaactgttc
     2401 atccagttag tacaaaccta taatcagttg tttgaagaga accctataaa tgcaagtggc
     2461 gtggatgcga aggctattct tagcgcccgc ctctctaaat cccgacggct agaaaacctg
     2521 atcgcacaat tacccggaga gaagaaaaat gggttgttcg gtaaccttat agcgctctca
     2581 ctaggcctga caccaaattt taagtcgaac ttcgacttag ctgaagatgc caaattgcag
     2641 cttagtaagg acacgtacga tgacgatctc gacaatctac tggcacaaat tggagatcag
     2701 tatgcggact tatttttggc tgccaaaaac cttagcgatg caatcctcct atctgacata
     2761 ctgagagtta atactgagat taccaaggcg ccgttatccg cttcaatgat caaaaggtac
     2821 gatgaacatc accaagactt gacacttctc aaggccctag tccgtcagca actgcctgag
     2881 aaatataagg aaatattctt tgatcagtcg aaaaacgggt acgcaggtta tattgacggc
     2941 ggagcgagtc aagaggaatt ctacaagttt atcaaaccca tattagagaa gatggatggg
     3001 acggaagagt tgcttgtaaa actcaatcgc gaagatctac tgcgaaagca gcggactttc
     3061 gacaacggta gcattccaca tcaaatccac ttaggcgaat tgcatgctat acttagaagg
     3121 caggaggatt tttatccgtt cctcaaagac aatcgtgaaa agattgagaa aatcctaacc
     3181 tttcgcatac cttactatgt gggacccctg gcccgaggga actctcggtt cgcatggatg
     3241 acaagaaagt ccgaagaaac gattactcca tggaattttg aggaagttgt cgataaaggt
     3301 gcgtcagctc aatcgttcat cgagaggatg accaactttg acaagaattt accgaacgaa
     3361 aaagtattgc ctaagcacag tttactttac gagtatttca cagtgtacaa tgaactcacg
     3421 aaagttaagt atgtcactga gggcatgcgt aaacccgcct ttctaagcgg agaacagaag
     3481 aaagcaatag tagatctgtt attcaagacc aaccgcaaag tgacagttaa gcaattgaaa
     3541 gaggactact ttaagaaaat tgaatgcttc gattctgtcg agatctccgg ggtagaagat
     3601 cgatttaatg cgtcacttgg tacgtatcat gacctcctaa agataattaa agataaggac
     3661 ttcctggata acgaagagaa tgaagatatc ttagaagata tagtgttgac tcttaccctc
     3721 tttgaagatc gggaaatgat tgaggaaaga ctaaaaacat acgctcacct gttcgacgat
     3781 aaggttatga aacagttaaa gaggcgtcgc tatacgggct ggggacgatt gtcgcggaaa
     3841 cttatcaacg ggataagaga caagcaaagt ggtaaaacta ttctcgattt tctaaagagc
     3901 gacggcttcg ccaataggaa ctttatgcag ctgatccatg atgactcttt aaccttcaaa
     3961 gaggatatac aaaaggcaca ggtttccgga caaggggact cattgcacga acatattgcg
     4021 aatcttgctg gttcgccagc catcaaaaag ggcatactcc agacagtcaa agtagtggat
     4081 gagctagtta aggtcatggg acgtcacaaa ccggaaaaca ttgtaatcga gatggcacgc
     4141 gaaaatcaaa cgactcagaa ggggcaaaaa aacagtcgag agcggatgaa gagaatagaa
     4201 gagggtatta aagaactggg cagccagatc ttaaaggagc atcctgtgga aaatacccaa
     4261 ttgcagaacg agaaacttta cctctattac ctacaaaatg gaagggacat gtatgttgat
     4321 caggaactgg acataaaccg tttatctgat tacgacgtcg atcacattgt accccaatcc
     4381 tttttgaagg acgattcaat cgacaataaa gtgcttacac gctcggataa gaaccgaggg
     4441 aaaagtgaca atgttccaag cgaggaagtc gtaaagaaaa tgaagaacta ttggcggcag
     4501 ctcctaaatg cgaaactgat aacgcaaaga aagttcgata acttaactaa agctgagagg
     4561 ggtggcttgt ctgaacttga caaggccgga tttattaaac gtcagctcgt ggaaacccgc
     4621 caaatcacaa agcatgttgc acagatacta gattcccgaa tgaatacgaa atacgacgag
     4681 aacgataagc tgattcggga agtcaaagta atcactttaa agtcaaaatt ggtgtcggac
     4741 ttcagaaagg attttcaatt ctataaagtt agggagataa ataactacca ccatgcgcac
     4801 gacgcttatc ttaatgccgt cgtagggacc gcactcatta agaaataccc gaagctagaa
     4861 agtgagtttg tgtatggtga ttacaaagtt tatgacgtcc gtaagatgat cgcgaaaagc
     4921 gaacaggaga taggcaaggc tacagccaaa tacttctttt attctaacat tatgaatttc
     4981 tttaagacgg aaatcactct ggcaaacgga gagatacgca aacgaccttt aattgaaacc
     5041 aatggggaga caggtgaaat cgtatgggat aagggccggg acttcgcgac ggtgagaaaa
     5101 gttttgtcca tgccccaagt caacatagta aagaaaactg aggtgcagac cggagggttt
     5161 tcaaaggaat cgattcttcc aaaaaggaat agtgataagc tcatcgctcg taaaaaggac
     5221 tgggacccga aaaagtacgg tggcttcgat agccctacag ttgcctattc tgtcctagta
     5281 gtggcaaaag ttgagaaggg aaaatccaag aaactgaagt cagtcaaaga attattgggg
     5341 ataacgatta tggagcgctc gtcttttgaa aagaacccca tcgacttcct tgaggcgaaa
     5401 ggttacaagg aagtaaaaaa ggatctcata attaaactac caaagtatag tctgtttgag
     5461 ttagaaaatg gccgaaaacg gatgttggct agcgccggag agcttcaaaa ggggaacgaa
     5521 ctcgcactac cgtctaaata cgtgaatttc ctgtatttag cgtcccatta cgagaagttg
     5581 aaaggttcac ctgaagataa cgaacagaag caactttttg ttgagcagca caaacattat
     5641 ctcgacgaaa tcatagagca aatttcggaa ttcagtaaga gagtcatcct agctgatgcc
     5701 aatctggaca aagtattaag cgcatacaac aagcacaggg ataaacccat acgtgagcag
     5761 gcggaaaata ttatccattt gtttactctt accaacctcg gcgctccagc cgcattcaag
     5821 tattttgaca caacgataga tcgcaaacga tacacttcta ccaaggaggt gctagacgcg
     5881 acactgattc accaatccat cacgggatta tatgaaactc ggatagattt gtcacagctt
     5941 gggggtgacg ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct
     6001 ggacctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     6061 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     6121 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     6181 cccaccctcg tgaccaccct gacctatgga gtgcagtgct tcagccgcta ccccgaccac
     6241 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     6301 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     6361 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     6421 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     6481 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     6541 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     6601 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     6661 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     6721 aagtctggtg gttctcccaa gaagaagagg aaagtctaac cggtcatcat caccatcacc
     6781 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     6841 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     6901 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     6961 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     7021 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgata ccgtcgacct
     7081 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     7141 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctag ggtgcctaat
     7201 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7261 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7321 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7381 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7441 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7501 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7561 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7621 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7681 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7741 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7801 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7861 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7921 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7981 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8041 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     8101 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8161 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8221 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8281 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8341 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8401 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8461 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8521 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8581 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8641 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8701 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8761 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8821 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8881 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8941 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9001 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9061 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9121 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9181 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9241 cccgaaaagt gccacctgac gtcgacggat cgggagatcg atctcccgat cccctagggt
     9301 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     9361 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     9421 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     9481 tacgggccag atatacgcgt t
//
