LOCUS       sequence-407295-       17618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ubiquitous expression of dCBEevoCDA1 in Drosophila.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17618)
  AUTHORS   Doll RM, Boutros M, Port F
  TITLE     A temperature-tolerant CRISPR base editor mediates highly efficient 
            and precise gene editing in Drosophila.
  JOURNAL   Sci Adv. 2023 Sep;9(35):eadj1568. doi: 10.1126/sciadv.adj1568. Epub 
            2023 Aug 30.
  PUBMED    37647411
REFERENCE   2  (bases 1 to 17618)
  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: "Sci Adv."; 
            date: "2023-09"; volume: "9(35):eadj1568. doi"; pages: " 
            10.1126/sciadv.adj1568. Epub 2023 Aug 30"
COMMENT     Sequence Label: pAct-dCBEevoCDA1
FEATURES             Location/Qualifiers
     source          1..17618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4239..4259
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4980..5003,5004..5006,5007..9080)
                     /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             4980..9080
                     /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             9111..9359
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9402..9422
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          9521..9586
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9716..9736
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10008..10142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10045..10064)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10099..10118
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    10171..10240
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(10459..10477)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10498..10514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10498..10514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10511..10533)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10522..10538
                     /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(10546..10552,10553..10570,10571..10576))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10591..10612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10729..10746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10900..11488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10980..10999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11659..12450,12451..12519))
                     /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     12282..12301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12520..12624)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(12650..13105)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12787..12808)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12999..13018
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13232..13254
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     13246..13263
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     13247..13263
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13270..13289
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        13270..13288
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(13322..13340)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(13792..13863,14265..14538,14613..15267,15329..15644,
                     15848..15979,16050..16664)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    17439..17472
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(17543..17618,1..2435)
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 agatctcgct gcctgttatg tggcccacaa accaagacac gttttatggc cattaaagct
       61 ggctgatcgt cgccaaacac caaatacata atgaatatgt acacattcga gaaagaagcg
      121 atcaaagaag cgtcttcggg cggagtagga gaatgcggag gagaaggaga acgagctgat
      181 ctagtatctc tccacaatcc aatgccaact gaccaactgg ccatattcgg agcaatttga
      241 agccaatttc catcgcctgg cgatcgctcc attcttggct atatgttttt caccgttacc
      301 cggggccatt ttcaaagact cgtcggcaag ataagattgt gtcactcgct gtctctcttc
      361 atttgtcgaa gaatgctgag gaatttcgcg atgacgtcgg cgagtatttt gaagaatgag
      421 aataatttgt atttatacga aaatcagtta gtggaatttt ctacaaaaac atgttatcta
      481 tagataattt tgttgcaaaa tatgttgact atgacaaaga ttgtatgtat atacctttaa
      541 tgtattctca ttttcttatg tatttataat ggcaatgatg atactgatga tattttaaga
      601 tgatgccaga ccaaaaggct tgaatttctg cgtcttttgc cgaacgcagt gcatgtgcaa
      661 ttgttgtttt ttggaatatt caattttcgg actgtccgct ttgatttcag tttcttggct
      721 tattcaaaaa gcaaagtaaa gccaaaaaag cgagatggca ataccaaatg cggcaaaacg
      781 gtagtggaag gaaaggggtg cggggcagcg gaaggaaggg tggggcgggg cgtggcgggg
      841 tctgtggctg ggcgcgacgt caccgacgtt ggagccactc ctttgaccat gtgtgcgtgt
      901 gtgtattatt cgtgtctcgc cactcgccgg ttgttttttt ctttttatgc tgcgctctct
      961 ctagcgccat ctcgcttacg catgctcaac gcaccgcatg ttgccgtttc cttttatgcg
     1021 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
     1081 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
     1141 aagcaacaac acaggtggct ttccctcttt cactttttgt ttgcaagccg cgtgcgagca
     1201 agacggcacg accggcaaac gcaattacgc tgacaaagag cagacgaagt tttggcgaaa
     1261 aacatcaagg cgcctgatac gaatgcattt gcaataacaa ttgcgatatt taatattgtt
     1321 tatgaagctg tttgacttca aaacacacaa aaaaaaaaaa taaaacaaat tatttgaaag
     1381 agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta ctgagtcaca
     1441 gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag cagcttagac
     1501 cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa cgcatagact
     1561 gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat tactattcaa
     1621 aactattaag atagcaatca cattcaatag ccaaatacta taccacctga gcgatgcaac
     1681 gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta tagaaatgct
     1741 tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt aaaaccggct
     1801 taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg tgttgcacaa
     1861 aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc taatatgaat
     1921 attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca ctataaacac
     1981 atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt gtaaaagcac
     2041 cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta acccccaaat
     2101 tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg cgatatacaa
     2161 gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg aaaacccaat
     2221 cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga gagcggagag
     2281 cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct gcgggaccag
     2341 ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc gacacaaagc
     2401 cgctccatca gccagcagtc gtctaatcca gagacaccaa accgaaagac ttaatttata
     2461 tttatttaat taattttaat aaaacacacc aaatgtaagt agctttcccc ttcccaacaa
     2521 caaaacacca tcgaaccact cccaccaaga aaaagcaata atcgagaaaa gccgcggaaa
     2581 atgtgtgatt ttttttgtaa acaaaacatt tttttatgtg ccagtgctga aagtgatcaa
     2641 aaaatactag ccacgagcta aagagttatt gtattgacca aaactccaaa aatacccaag
     2701 tttggcccta aattgtcaat caaaatacca ataggtcgaa agacatcaaa attaacaaaa
     2761 ccagggtttc aaataccata actcaagaat caggattaca actgcagatt tcaggatata
     2821 tacatacaaa ttatacgaaa ttataaaaac caaagcaatt caatagcccc aactcaaatg
     2881 ttaggatcta atatagtgtt taaagccaag ctcgctgatg tgggcgtgtc acgatttcac
     2941 ccaaagatat gccaaattac gaattgcaaa tcaattcgcc aacactttct ttttttccca
     3001 cgccctaaaa caccagatca tccataaatg tacatacata cagtatatgc atattataat
     3061 ctgtaaaact agatcaggtt cttgaaaata gtgacgtagg cagccgtttt ggctgaagca
     3121 gaaatttttg ccggtttttc aaagttgtag ttgcaaaaat ggagaaaacc ttcgagcatt
     3181 cgttcatata cacacactca cgcgcaaaat aacgagagag agtgtatgtg tgtgtgagag
     3241 agcgaaagcc agacgacggt ttgcttttcg cctcgaaaca tgaccatata tggtcacaaa
     3301 acttggccgc cgcaattcaa cacaccagcg ctctccttcg cacccatagc gaccatggcg
     3361 cggagcgagc gagatggcga gagcgagcga cgcctatggc gacgtcgacg caggcagcga
     3421 ttgaaaaacg cagttaactg gcattcaaca ttcaccagcc actttcagtc ggtttattcc
     3481 agtcattcct ttcaaaccgt gcggtcgctt agctcagcct cgccacttgc gtttacagta
     3541 gttttcacgc cttgaatttg ttaaatcgaa caaaaaggta aagtttaact agctttgaaa
     3601 agtttcgtgg ctcttaattg ttaaattttc tagagtgcgt ttagtgtttt tttttttttt
     3661 tattttgtaa tgttaatttc gggttccaat tcgagtttta ggcagccgcc attttaaggg
     3721 cgcatacaca caggcaactg tgctctcttt gcggctttct tttgcaccgg cattcgttaa
     3781 gtgtcgtcta gaagcttctc ccctcccttt tcggcatatt cgtattgtgg ttttaatttt
     3841 tcggggcggg gccttctatt ttgtaactgt tcttttaatt tcttattaca attcgatcgc
     3901 aagtgaaaat cagttttcaa tcggaaaagt atttttttat gaaatttttt tttgtccaag
     3961 attaaaattt tgtactaaaa aaacgtacat tgcattgagt gatttttaat tgtacacgaa
     4021 aaacaagtta gtttgttatg acaattgtac tttggtagac cagcgcagtc caaggaaacc
     4081 acgcaaattc tcagtttttt ttttgccatt tctacattac caaataaggt aaccaaaaac
     4141 taatgggaaa tccgcattct ttccattgca gcttacagga tcgatccccg ggaattccca
     4201 ccatgaaacg gacagccgac ggaagcgagt tcgagtcacc aaagaagaag cggaaagtca
     4261 gtaccgacgc cgagtacgtg cggatccacg agaagctgga tatctataca ttcaagaagc
     4321 agtttagcaa caataagaag tccgtgtctc acagatgcta cgtgctgttc gagctgaagc
     4381 ggagaggaga gaggcgcgcc tgtttttggg gctatgccgt gaacaagcca cagtctggaa
     4441 ccgagagggg aatccacgca gagatcttca gcatcaggaa ggtggaggag tacctgcgcg
     4501 acaaccccgg ccagtttaca atcaattggt atagctcctg gagcccttgc gccgattgtg
     4561 ccgagaagat cctggagtgg tacaaccagg agctgagggg caatggccac accctgaaga
     4621 tctgggtgtg caagctgtac tatgagaaga acgccaggaa tcagatcggc ctgtggaacc
     4681 tgcgcgacaa tggcgtgggc ctgaacgtga tggtgtccga gcactatcag tgctgtcgca
     4741 agatctttat ccagtctagc cacaatcagc tgaacgagaa tcggtggctg gagaagacac
     4801 tgaagagagc cgagaagcgg agaagcgagc tgtccatcat gtttcaggtg aagatcctgc
     4861 acaccacaaa gtctcccgcc gtgtctggcg gatctagcgg aggatcctct ggcagcgaga
     4921 caccaggaac aagcgagtca gcaacaccag agagcagtgg cggcagcagc ggcggcagcg
     4981 acaagaagta ctccattggg ctcgccatcg gcacaaacag cgtcggctgg gccgtcatta
     5041 cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc gatcgccaca
     5101 gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg gccgaagcca
     5161 cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg atctgctacc
     5221 tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc cataggctgg
     5281 aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc tttggcaata
     5341 tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg aggaagaagc
     5401 ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg gcgcatatga
     5461 tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac agcgatgtcg
     5521 acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag aacccgatca
     5581 acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc
     5641 tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt ggtaatctta
     5701 tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg gccgaagatg
     5761 ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg ctggcccaga
     5821 tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac gccattctgc
     5881 tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc gctagtatga
     5941 tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt gtcagacagc
     6001 aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc tacgccggat
     6061 acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc atcttggaaa
     6121 aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg ttgcgcaaac
     6181 agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa ctgcacgcta
     6241 tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa aagattgaga
     6301 aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga aattccagat
     6361 tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc gaggaagtcg
     6421 tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt gataaaaatc
     6481 tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc acagtttata
     6541 acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca ttcctgtctg
     6601 gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa gttaccgtga
     6661 aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg
     6721 gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg aaaatcatta
     6781 aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac attgtcctca
     6841 cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact tacgctcatc
     6901 tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga tgggggcggc
     6961 tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca atcctggatt
     7021 ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat gatgactctc
     7081 tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac agtcttcacg
     7141 agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg cagaccgtta
     7201 aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat atcgttatcg
     7261 agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg gaaaggatga
     7321 agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa cacccagttg
     7381 aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac ggcagggaca
     7441 tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg gatcatatcg
     7501 tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca agatccgata
     7561 aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt
     7621 attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat aatctgacta
     7681 aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa aggcagcttg
     7741 ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc atgaacacca
     7801 agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg aagtctaagc
     7861 tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc aacaattacc
     7921 accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc
     7981 ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt aggaaaatga
     8041 tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt tacagcaata
     8101 ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg aagcgaccac
     8161 ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg gatttcgcga
     8221 cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc gaagtacaga
     8281 ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac
     8341 gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca gtcgcttaca
     8401 gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg
     8461 aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc atcgactttc
     8521 tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt cccaagtact
     8581 ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga
     8641 aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg gccagccact
     8701 atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc gtggaacaac
     8761 acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa agagtgatcc
     8821 tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg gataagccca
     8881 tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg ggcgcgcctg
     8941 cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct acaaaggagg
     9001 tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca agaatcgacc
     9061 tctctcagct cggtggagac agcggcggtt ccggtggatc cggcggaagc acgaacctga
     9121 gtgatataat tgaaaaggaa accggtaaac agctcgttat ccaggagtcg atcctcatgc
     9181 tgcccgagga agtcgaggaa gtcatcggta acaaaccgga atcggatatt ctggtacata
     9241 cggcctatga tgaatccacc gatgagaatg taatgttgtt gacatcggat gcaccggagt
     9301 ataagccttg ggcgctcgtc atccaggact ccaatggcga gaacaagatt aagatgctct
     9361 ccggaggatc caaacgaaca gctgacggta gtgaattcga accaaaaaaa aagcgcaagg
     9421 tataactcga gggtacctct agaggatctt tgtgaaggaa ccttacttct gtggtgtgac
     9481 ataattggac aaactaccta cagagattta aagctctaag gtaaatataa aatttttaag
     9541 tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc caacctatgg
     9601 aactgatgaa tgggagcagt ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga
     9661 agaaatgcca tctagtgatg atgaggctac tgctgactct caacattcta ctcctccaaa
     9721 aaagaagaga aaggtagaag accccaagga ctttccttca gaattgctaa gttttttgag
     9781 tcatgctgtg tttagtaata gaactcttgc ttgctttgct atttacacca caaaggaaaa
     9841 agctgcactg ctatacaaga aaattatgga aaaatatttg atgtatagtg ccttgactag
     9901 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     9961 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
    10021 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
    10081 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
    10141 tccactagtg tcgacgatgt aggtcacggt ctcgaagccg cggtgcgggt gccagggcgt
    10201 gcccttgggc tccccgggcg cgtactccac ctcacccatc tggtccatca tgatgaacgg
    10261 gtcgaggtgg cggtagttga tcccggcgaa cgcgcggcgc accgggaagc cctcgccctc
    10321 gaaaccgctg ggcgcggtgg tcacggtgag cacgggacgt gcgacggcgt cggcgggtgc
    10381 ggatacgcgg ggcagcgtca gcgggttctc gacggtcacg gcgggcatgt cgacactagt
    10441 tctagccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc
    10501 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    10561 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    10621 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    10681 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    10741 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    10801 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    10861 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    10921 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    10981 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    11041 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    11101 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    11161 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    11221 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    11281 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    11341 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    11401 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    11461 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    11521 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
    11581 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    11641 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    11701 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    11761 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    11821 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    11881 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    11941 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    12001 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    12061 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    12121 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    12181 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    12241 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    12301 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    12361 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    12421 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    12481 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    12541 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    12601 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
    12661 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
    12721 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
    12781 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
    12841 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
    12901 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
    12961 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
    13021 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
    13081 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
    13141 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
    13201 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
    13261 agtgaattgt aatacgactc actatagggc gaattgggta cgtaccgggc ccctagtatg
    13321 tatgtaagtt aataaaaccc atttttgcgg aaagtagata aaaaaaacat tttttttttt
    13381 tactgcactg gatatcattg aacttatctg atcagtttta aatttacttc gatccaaggg
    13441 tatttgatgt accaggttct ttcgattacc tctcactcaa aatgacattc cactcaaagt
    13501 cagcgctgtt tgcctccttc tctgtccaca gaaatatcgc cgtctctttc gccgctgcgt
    13561 ccgctatctc tttcgccacc gtttgtagcg ttacgtagcg tcaatgtccg ccttcagttg
    13621 cattttgtca gcggtttcgt gacgaagctc caagcggttt acgccatcaa ttaaacacaa
    13681 agtgctgtgc caaaactcct ctcgcttctt atttttgttt gttttttgag tgattggggt
    13741 ggtgattggt tttgggtggg taagcagggg aaagtgtgaa aaatcccggc aatgggccaa
    13801 gaggatcagg agctattaat tcgcggaggc agcaaacacc catctgccga gcatctgaac
    13861 aatgtgagta gtacatgtgc atacatctta agttcacttg atctatagga actgcgattg
    13921 caacatcaaa ttgtctgcgg cgtgagaact gcgacccaca aaaatcccaa accgcaattg
    13981 cacaaacaaa tagtgacacg aaacagatta ttctggtagc tgttctcgct atataagaca
    14041 atttttgaga tcatatcatg atcaagacat ctaaaggcat tcattttcga ctatattctt
    14101 ttttacaaaa aatataacaa ccagatattt taagctgatc ctagatgcac aaaaaataaa
    14161 taaaagtata aacctacttc gtaggatact tcggggtact ttttgttcgg ggttagatga
    14221 gcataacgct tgtagttgat atttgagatc ccctatcatt gcagggtgac agcggagcgg
    14281 cttcgcagag ctgcattaac cagggcttcg ggcaggccaa aaactacggc acgctccggc
    14341 cacccagtcc gccggaggac tccggttcag ggagcggcca actagccgag aacctcacct
    14401 atgcctggca caatatggac atctttgggg cggtcaatca gccgggctcc ggatggcggc
    14461 agctggtcaa ccggacacgc ggactattct gcaacgagcg acacataccg gcgcccagga
    14521 aacatttgct caagaacggt gagtttctat tcgcagtcgg ctgatctgtg tgaaatctta
    14581 ataaagggtc caattaccaa tttgaaactc agtttgcggc gtggcctatc cgggcgaact
    14641 tttggccgtg atgggcagtt ccggtgccgg aaagacgacc ctgctgaatg cccttgcctt
    14701 tcgatcgccg cagggcatcc aagtatcgcc atccgggatg cgactgctca atggccaacc
    14761 tgtggacgcc aaggagatgc aggccaggtg cgcctatgtc cagcaggatg acctctttat
    14821 cggctcccta acggccaggg aacacctgat tttccaagcc atggtgcgga tgccacgaca
    14881 tctgacctat cggcagcgag tggcccgcgt ggatcaggtg atccaggagc tttcgctcag
    14941 caaatgtcag cacacgatca tcggtgtgcc cggcagggtg aaaggtctgt ccggcggaga
    15001 aaggaagcgt ctggcattcg cctccgaggc tctaaccgat ccgccgcttc tgatctgcga
    15061 tgagcccacc tccggactgg actcctttac cgcccacagc gtcgtccagg tgctgaagaa
    15121 gctgtcgcag aagggcaaga ccgtcatcct gaccattcat cagccgtctt ccgagctgtt
    15181 tgagctcttt gacaagatcc ttctgatggc cgagggcagg gtagctttct tgggcactcc
    15241 cagcgaagcc gtcgacttct tttcctagtg agttcgatgt gtttattaag ggtatctagt
    15301 attacataac atctcaactc ctatccagcg tgggtgccca gtgtcctacc aactacaatc
    15361 cggcggactt ttacgtacag gtgttggccg ttgtgcccgg acgggagatc gagtcccgtg
    15421 atcggatcgc caagatatgc gacaattttg ccattagcaa agtagcccgg gatatggagc
    15481 agttgttggc caccaaaaat ctggagaagc cactggagca gccggagaat gggtacacct
    15541 acaaggccac ctggttcatg cagttccggg cggtcctgtg gcgatcctgg ctgtcggtgc
    15601 tcaaggaacc actcctcgta aaagtgcgac ttattcagac aacggtgagt ggttccagtg
    15661 gaaacaaatg atataacgct tacaattctt ggaaacaaat tcgctagatt ttagatagaa
    15721 ttgcctgatt ccacaccctt cttagttttt ttcaatgaga tgtatagttt atagttttgc
    15781 agaagataaa taaatttcat ttaactcgcg aatattaatg agatgcgagt aacattttaa
    15841 tttgcagatg gttgccatct tgattggcct catctttttg ggccaacaac tcacgcaagt
    15901 gggtgtgatg aatatcaacg gagccatctt cctcttcctg accaacatga cctttcaaaa
    15961 cgtctttgcc acgataaatg taagtcatgt ttagaataca tttgcatttc aataatttac
    16021 taactttcta atgaatcgat tcgatttagg tgttcacctc agagctgcca gtttttatga
    16081 gggaggcccg aagtcgactt tatcgctgtg acacatactt tctgggcaaa acgattgccg
    16141 aattgccgct ttttctcaca gtgccactgg tcttcacggc gattgcctat ccgatgatcg
    16201 gactgcgggc cggagtgctg cacttcttca actgcctggc gctggtcact ctggtggcca
    16261 atgtgtcaac gtccttcgga tatctaatat cctgcgccag ctcctcgacc tcgatggcgc
    16321 tgtctgtggg tccgccggtt atcataccat tcctgctctt tggcggcttc ttcttgaact
    16381 cgggctcggt gccagtatac ctcaaatggt tgtcgtacct ctcatggttc cgttacgcca
    16441 acgagggtct gctgattaac caatgggcgg acgtggagcc gggcgaaatt agctgcacat
    16501 cgtcgaacac cacgtgcccc agttcgggca aggtcatcct ggagacgctt aacttctccg
    16561 ccgccgatct gccgctggac tacgtgggtc tggccattct catcgtgagc ttccgggtgc
    16621 tcgcatatct ggctctaaga cttcgggccc gacgcaagga gtagccgaca tatatccgaa
    16681 ataactgctt gttttttttt tttaccatta ttaccatcgt gtttactgtt tattgccccc
    16741 tcaaaaagct aatgtaatta tatttgtgcc aataaaaaca agatatgacc tatagaatac
    16801 aagtatttcc ccttcgaaca tccccacaag tagactttgg atttgtcttc taaccaaaag
    16861 acttacacac ctgcatacct tacatcaaaa actcgtttat cgctacataa aacaccggga
    16921 tatatttttt atatacatac ttttcaaatc gcgcgccctc ttcataattc acctccacca
    16981 caccacgttt cgtagttgct ctttcgctgt ctcccacccg ctctccgcaa cacattcacc
    17041 ttttgttcga cgaccttgga gcgactgtcg ttagttccgc gcgattcggt tcgctcaaat
    17101 ggttccgagt ggttcatttc gtctcaatag aaattagtaa taaatatttg tatgtacaat
    17161 ttatttgctc caatatattt gtatatattt ccctcacagc tatatttatt ctaatttaat
    17221 attatgactt tttaaggtaa ttttttgtga cctgttcgga gtgattagcg ttacaatttg
    17281 aactgaaagt gacatccagt gtttgttcct tgtgtagatg catctcaaaa aaatggtggg
    17341 cataatagtg ttgtttatat atatcaaaaa taacaactat aataataaga atacatttaa
    17401 tttagaaaat gcttggattt cactggaact aggctagcat aacttcgtat aatgtatgct
    17461 atacgaagtt atgctagcgg atccaagctt gcggccgcaa ttctatattc taaaaacaca
    17521 aatgatactt ctaaaaaaaa atcatgaatg gcatcaactc tgaatcaaat ctttgcagat
    17581 gcacctactt ctcatttcca ctgtcacatc atttttcc
//
