LOCUS       sequence-407293-       17680 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ubiquitous expression of dCBEevoAPOBEC1 in Drosophila.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17680)
  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 17680)
  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-dCBEevoAPOBEC1
FEATURES             Location/Qualifiers
     source          1..17680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             282..302
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1083..1109,1110..1112,1113..5186)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1083..5186
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5217..5465
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5508..5528
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5627..5692
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5822..5842
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6114..6248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6151..6170)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6205..6224
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    6277..6346
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(6565..6583)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6604..6620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6604..6620)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6617..6639)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6628..6644
                     /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(6652..6658,6659..6676,6677..6682))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6697..6718
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6835..6852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7006..7594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7086..7105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7765..8556,8557..8625))
                     /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     8388..8407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8626..8730)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8756..9211)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8893..8914)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9105..9124
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9338..9360
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9352..9369
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9353..9369
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9376..9395
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9376..9394
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9428..9446)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(9898..9969,10371..10644,10719..11373,11435..11750,
                     11954..12085,12156..12770)
                     /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    13545..13578
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        13649..16159
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 caagattaaa attttgtact aaaaaaacgt acattgcatt gagtgatttt taattgtaca
       61 cgaaaaacaa gttagtttgt tatgacaatt gtactttggt agaccagcgc agtccaagga
      121 aaccacgcaa attctcagtt ttttttttgc catttctaca ttaccaaata aggtaaccaa
      181 aaactaatgg gaaatccgca ttctttccat tgcagcttac aggatcgatc cccggaattc
      241 ccaccatgaa acggacagcc gacggaagcg agttcgagtc accaaagaag aagcggaaag
      301 tcagttcaaa gactgggcct gtcgccgtcg atccaaccct gcgccgccgg attgaacctc
      361 acgagtttga agtgttcttt gacccccggg agctgagaaa ggagacatgc ctgctgtacg
      421 agatcaactg gggaggcagg cactccatct ggaggcacac ctctcagaac acaaataagc
      481 acgtggaggt gaacttcatc gagaagttta ccacagagcg gtacttctgc cccaatacca
      541 gatgtagcat cacatggttt ctgagctggt ccccttgcgg agagtgtagc agggccatca
      601 ccgagttcct gtccagatat ccaaatgtga cactgtttat ctacatcgcc aggctgtatc
      661 acctggcaaa cccaaggaat aggcagggcc tgcgcgatct gatcagctcc ggcgtgacca
      721 tccagatcat gacagagcag gagtccggct actgctggca caacttcgtg aattattctc
      781 ctagcaacga gtcccactgg cctaggtacc cacacctgtg ggtgcgcctg tacgtgctgg
      841 agctgtattg catcatcctg ggcctgcccc cttgtctgaa tatcctgcgg agaaagcaga
      901 gccagctgac ctcctttaca atcgccctgc agtcttgtca ctatcagagg ctgccacccc
      961 acatcctgtg ggccacaggc ctgaagtctg gcggatctag cggaggatcc tctggcagcg
     1021 agacaccagg aacaagcgag tcagcaacac cagagagcag tggcggcagc agcggcggca
     1081 gcatggacaa gaagtactcc attgggctcg ccatcggcac aaacagcgtc ggctgggccg
     1141 tcattacgga cgagtacaag gtgccgagca aaaaattcaa agttctgggc aataccgatc
     1201 gccacagcat aaagaagaac ctcattggcg ccctcctgtt cgactccggg gagacggccg
     1261 aagccacgcg gctcaaaaga acagcacggc gcagatatac ccgcagaaag aatcggatct
     1321 gctacctgca ggagatcttt agtaatgaga tggctaaggt ggatgactct ttcttccata
     1381 ggctggagga gtcctttttg gtggaggagg ataaaaagca cgagcgccac ccaatctttg
     1441 gcaatatcgt ggacgaggtg gcgtaccatg aaaagtaccc aaccatatat catctgagga
     1501 agaagcttgt agacagtact gataaggctg acttgcggtt gatctatctc gcgctggcgc
     1561 atatgatcaa atttcgggga cacttcctca tcgaggggga cctgaaccca gacaacagcg
     1621 atgtcgacaa actctttatc caactggttc agacttacaa tcagcttttc gaagagaacc
     1681 cgatcaacgc atccggagtt gacgccaaag caatcctgag cgctaggctg tccaaatccc
     1741 ggcggctcga aaacctcatc gcacagctcc ctggggagaa gaagaacggc ctgtttggta
     1801 atcttatcgc cctgtcactc gggctgaccc ccaactttaa atctaacttc gacctggccg
     1861 aagatgccaa gcttcaactg agcaaagaca cctacgatga tgatctcgac aatctgctgg
     1921 cccagatcgg cgaccagtac gcagaccttt ttttggcggc aaagaacctg tcagacgcca
     1981 ttctgctgag tgatattctg cgagtgaaca cggagatcac caaagctccg ctgagcgcta
     2041 gtatgatcaa gcgctatgat gagcaccacc aagacttgac tttgctgaag gcccttgtca
     2101 gacagcaact gcctgagaag tacaaggaaa ttttcttcga tcagtctaaa aatggctacg
     2161 ccggatacat tgacggcgga gcaagccagg aggaatttta caaatttatt aagcccatct
     2221 tggaaaaaat ggacggcacc gaggagctgc tggtaaagct taacagagaa gatctgttgc
     2281 gcaaacagcg cactttcgac aatggaagca tcccccacca gattcacctg ggcgaactgc
     2341 acgctatcct caggcggcaa gaggatttct accccttttt gaaagataac agggaaaaga
     2401 ttgagaaaat cctcacattt cggataccct actatgtagg ccccctcgcc cggggaaatt
     2461 ccagattcgc gtggatgact cgcaaatcag aagagaccat cactccctgg aacttcgagg
     2521 aagtcgtgga taagggggcc tctgcccagt ccttcatcga aaggatgact aactttgata
     2581 aaaatctgcc taacgaaaag gtgcttccta aacactctct gctgtacgag tacttcacag
     2641 tttataacga gctcaccaag gtcaaatacg tcacagaagg gatgagaaag ccagcattcc
     2701 tgtctggaga gcagaagaaa gctatcgtgg acctcctctt caagacgaac cggaaagtta
     2761 ccgtgaaaca gctcaaagaa gactatttca aaaagattga atgtttcgac tctgttgaaa
     2821 tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa
     2881 tcattaaaga caaggacttc ctggacaatg aggagaacga ggacattctt gaggacattg
     2941 tcctcaccct tacgttgttt gaagataggg agatgattga agaacgcttg aaaacttacg
     3001 ctcatctctt cgacgacaaa gtcatgaaac agctcaagag gcgccgatat acaggatggg
     3061 ggcggctgtc aagaaaactg atcaatggga tccgagacaa gcagagtgga aagacaatcc
     3121 tggattttct taagtccgat ggatttgcca accggaactt catgcagttg atccatgatg
     3181 actctctcac ctttaaggag gacatccaga aagcacaagt ttctggccag ggggacagtc
     3241 ttcacgagca catcgctaat cttgcaggta gcccagctat caaaaaggga atactgcaga
     3301 ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag gcataagccc gagaatatcg
     3361 ttatcgagat ggcccgagag aaccaaacta cccagaaggg acagaagaac agtagggaaa
     3421 ggatgaagag gattgaagag ggtataaaag aactggggtc ccaaatcctt aaggaacacc
     3481 cagttgaaaa cacccagctt cagaatgaga agctctacct gtactacctg cagaacggca
     3541 gggacatgta cgtggatcag gaactggaca tcaatcggct ctccgactac gacgtggatc
     3601 atatcgtgcc ccagtctttt ctcaaagatg attctattga taataaagtg ttgacaagat
     3661 ccgataaaaa tagagggaag agtgataacg tcccctcaga agaagttgtc aagaaaatga
     3721 aaaattattg gcggcagctg ctgaacgcca aactgatcac acaacggaag ttcgataatc
     3781 tgactaaggc tgaacgaggt ggcctgtctg agttggataa agccggcttc atcaaaaggc
     3841 agcttgttga gacacgccag atcaccaagc acgtggccca aattctcgat tcacgcatga
     3901 acaccaagta cgatgaaaat gacaaactga ttcgagaggt gaaagttatt actctgaagt
     3961 ctaagctggt ctcagatttc agaaaggact ttcagtttta taaggtgaga gagatcaaca
     4021 attaccacca tgcgcatgat gcctacctga atgcagtggt aggcactgca cttatcaaaa
     4081 aatatcccaa gcttgaatct gaatttgttt acggagacta taaagtgtac gatgttagga
     4141 aaatgatcgc aaagtctgag caggaaatag gcaaggccac cgctaagtac ttcttttaca
     4201 gcaatattat gaattttttc aagaccgaga ttacactggc caatggagag attcggaagc
     4261 gaccacttat cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag ggtagggatt
     4321 tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag
     4381 tacagaccgg aggcttctcc aaggaaagta tcctcccgaa aaggaacagc gacaagctga
     4441 tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg attcgattct cctacagtcg
     4501 cttacagtgt actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg
     4561 tcaaggaact gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg
     4621 actttctcga ggcgaaagga tataaagagg tcaaaaaaga cctcatcatt aagcttccca
     4681 agtactctct ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc
     4741 tgcagaaagg taacgagctg gcactgccct ctaaatacgt taatttcttg tatctggcca
     4801 gccactatga aaagctcaaa gggtctcccg aagataatga gcagaagcag ctgttcgtgg
     4861 aacaacacaa acactacctt gatgagatca tcgagcaaat aagcgaattc tccaaaagag
     4921 tgatcctcgc cgacgctaac ctcgataagg tgctttctgc ttacaataag cacagggata
     4981 agcccatcag ggagcaggca gaaaacatta tccacttgtt tactctgacc aacttgggcg
     5041 cgcctgcagc cttcaagtac ttcgacacca ccatagacag aaagcggtac acctctacaa
     5101 aggaggtcct ggacgccaca ctgattcatc agtcaattac ggggctctat gaaacaagaa
     5161 tcgacctctc tcagctcggt ggagacagcg gcggttccgg tggatccggc ggaagcacga
     5221 acctgagtga tataattgaa aaggaaaccg gtaaacagct cgttatccag gagtcgatcc
     5281 tcatgctgcc cgaggaagtc gaggaagtca tcggtaacaa accggaatcg gatattctgg
     5341 tacatacggc ctatgatgaa tccaccgatg agaatgtaat gttgttgaca tcggatgcac
     5401 cggagtataa gccttgggcg ctcgtcatcc aggactccaa tggcgagaac aagattaaga
     5461 tgctctccgg aggatccaaa cgaacagctg acggtagtga attcgaacca aaaaaaaagc
     5521 gcaaggtata actcgagggt acctctagag gatctttgtg aaggaacctt acttctgtgg
     5581 tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt
     5641 tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt agattccaac
     5701 ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa acctgttttg
     5761 ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac attctactcc
     5821 tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat tgctaagttt
     5881 tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt acaccacaaa
     5941 ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tatttgatgt atagtgcctt
     6001 gactagagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     6061 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     6121 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6181 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6241 tctggatcca ctagtgtcga cgatgtaggt cacggtctcg aagccgcggt gcgggtgcca
     6301 gggcgtgccc ttgggctccc cgggcgcgta ctccacctca cccatctggt ccatcatgat
     6361 gaacgggtcg aggtggcggt agttgatccc ggcgaacgcg cggcgcaccg ggaagccctc
     6421 gccctcgaaa ccgctgggcg cggtggtcac ggtgagcacg ggacgtgcga cggcgtcggc
     6481 gggtgcggat acgcggggca gcgtcagcgg gttctcgacg gtcacggcgg gcatgtcgac
     6541 actagttcta gccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc
     6601 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6661 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6721 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6781 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6841 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6901 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6961 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7021 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7081 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7141 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7201 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7261 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7321 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7381 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7441 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7501 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7561 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7621 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7681 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7741 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7801 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7861 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7921 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7981 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8041 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8101 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8161 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8221 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8281 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8341 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8401 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8461 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8521 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8581 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8641 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8701 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     8761 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     8821 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     8881 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     8941 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     9001 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     9061 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     9121 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     9181 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     9241 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     9301 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     9361 acggccagtg aattgtaata cgactcacta tagggcgaat tgggtacgta ccgggcccct
     9421 agtatgtatg taagttaata aaacccattt ttgcggaaag tagataaaaa aaacattttt
     9481 tttttttact gcactggata tcattgaact tatctgatca gttttaaatt tacttcgatc
     9541 caagggtatt tgatgtacca ggttctttcg attacctctc actcaaaatg acattccact
     9601 caaagtcagc gctgtttgcc tccttctctg tccacagaaa tatcgccgtc tctttcgccg
     9661 ctgcgtccgc tatctctttc gccaccgttt gtagcgttac gtagcgtcaa tgtccgcctt
     9721 cagttgcatt ttgtcagcgg tttcgtgacg aagctccaag cggtttacgc catcaattaa
     9781 acacaaagtg ctgtgccaaa actcctctcg cttcttattt ttgtttgttt tttgagtgat
     9841 tggggtggtg attggttttg ggtgggtaag caggggaaag tgtgaaaaat cccggcaatg
     9901 ggccaagagg atcaggagct attaattcgc ggaggcagca aacacccatc tgccgagcat
     9961 ctgaacaatg tgagtagtac atgtgcatac atcttaagtt cacttgatct ataggaactg
    10021 cgattgcaac atcaaattgt ctgcggcgtg agaactgcga cccacaaaaa tcccaaaccg
    10081 caattgcaca aacaaatagt gacacgaaac agattattct ggtagctgtt ctcgctatat
    10141 aagacaattt ttgagatcat atcatgatca agacatctaa aggcattcat tttcgactat
    10201 attctttttt acaaaaaata taacaaccag atattttaag ctgatcctag atgcacaaaa
    10261 aataaataaa agtataaacc tacttcgtag gatacttcgg ggtacttttt gttcggggtt
    10321 agatgagcat aacgcttgta gttgatattt gagatcccct atcattgcag ggtgacagcg
    10381 gagcggcttc gcagagctgc attaaccagg gcttcgggca ggccaaaaac tacggcacgc
    10441 tccggccacc cagtccgccg gaggactccg gttcagggag cggccaacta gccgagaacc
    10501 tcacctatgc ctggcacaat atggacatct ttggggcggt caatcagccg ggctccggat
    10561 ggcggcagct ggtcaaccgg acacgcggac tattctgcaa cgagcgacac ataccggcgc
    10621 ccaggaaaca tttgctcaag aacggtgagt ttctattcgc agtcggctga tctgtgtgaa
    10681 atcttaataa agggtccaat taccaatttg aaactcagtt tgcggcgtgg cctatccggg
    10741 cgaacttttg gccgtgatgg gcagttccgg tgccggaaag acgaccctgc tgaatgccct
    10801 tgcctttcga tcgccgcagg gcatccaagt atcgccatcc gggatgcgac tgctcaatgg
    10861 ccaacctgtg gacgccaagg agatgcaggc caggtgcgcc tatgtccagc aggatgacct
    10921 ctttatcggc tccctaacgg ccagggaaca cctgattttc caagccatgg tgcggatgcc
    10981 acgacatctg acctatcggc agcgagtggc ccgcgtggat caggtgatcc aggagctttc
    11041 gctcagcaaa tgtcagcaca cgatcatcgg tgtgcccggc agggtgaaag gtctgtccgg
    11101 cggagaaagg aagcgtctgg cattcgcctc cgaggctcta accgatccgc cgcttctgat
    11161 ctgcgatgag cccacctccg gactggactc ctttaccgcc cacagcgtcg tccaggtgct
    11221 gaagaagctg tcgcagaagg gcaagaccgt catcctgacc attcatcagc cgtcttccga
    11281 gctgtttgag ctctttgaca agatccttct gatggccgag ggcagggtag ctttcttggg
    11341 cactcccagc gaagccgtcg acttcttttc ctagtgagtt cgatgtgttt attaagggta
    11401 tctagtatta cataacatct caactcctat ccagcgtggg tgcccagtgt cctaccaact
    11461 acaatccggc ggacttttac gtacaggtgt tggccgttgt gcccggacgg gagatcgagt
    11521 cccgtgatcg gatcgccaag atatgcgaca attttgccat tagcaaagta gcccgggata
    11581 tggagcagtt gttggccacc aaaaatctgg agaagccact ggagcagccg gagaatgggt
    11641 acacctacaa ggccacctgg ttcatgcagt tccgggcggt cctgtggcga tcctggctgt
    11701 cggtgctcaa ggaaccactc ctcgtaaaag tgcgacttat tcagacaacg gtgagtggtt
    11761 ccagtggaaa caaatgatat aacgcttaca attcttggaa acaaattcgc tagattttag
    11821 atagaattgc ctgattccac acccttctta gtttttttca atgagatgta tagtttatag
    11881 ttttgcagaa gataaataaa tttcatttaa ctcgcgaata ttaatgagat gcgagtaaca
    11941 ttttaatttg cagatggttg ccatcttgat tggcctcatc tttttgggcc aacaactcac
    12001 gcaagtgggt gtgatgaata tcaacggagc catcttcctc ttcctgacca acatgacctt
    12061 tcaaaacgtc tttgccacga taaatgtaag tcatgtttag aatacatttg catttcaata
    12121 atttactaac tttctaatga atcgattcga tttaggtgtt cacctcagag ctgccagttt
    12181 ttatgaggga ggcccgaagt cgactttatc gctgtgacac atactttctg ggcaaaacga
    12241 ttgccgaatt gccgcttttt ctcacagtgc cactggtctt cacggcgatt gcctatccga
    12301 tgatcggact gcgggccgga gtgctgcact tcttcaactg cctggcgctg gtcactctgg
    12361 tggccaatgt gtcaacgtcc ttcggatatc taatatcctg cgccagctcc tcgacctcga
    12421 tggcgctgtc tgtgggtccg ccggttatca taccattcct gctctttggc ggcttcttct
    12481 tgaactcggg ctcggtgcca gtatacctca aatggttgtc gtacctctca tggttccgtt
    12541 acgccaacga gggtctgctg attaaccaat gggcggacgt ggagccgggc gaaattagct
    12601 gcacatcgtc gaacaccacg tgccccagtt cgggcaaggt catcctggag acgcttaact
    12661 tctccgccgc cgatctgccg ctggactacg tgggtctggc cattctcatc gtgagcttcc
    12721 gggtgctcgc atatctggct ctaagacttc gggcccgacg caaggagtag ccgacatata
    12781 tccgaaataa ctgcttgttt ttttttttta ccattattac catcgtgttt actgtttatt
    12841 gccccctcaa aaagctaatg taattatatt tgtgccaata aaaacaagat atgacctata
    12901 gaatacaagt atttcccctt cgaacatccc cacaagtaga ctttggattt gtcttctaac
    12961 caaaagactt acacacctgc ataccttaca tcaaaaactc gtttatcgct acataaaaca
    13021 ccgggatata ttttttatat acatactttt caaatcgcgc gccctcttca taattcacct
    13081 ccaccacacc acgtttcgta gttgctcttt cgctgtctcc cacccgctct ccgcaacaca
    13141 ttcacctttt gttcgacgac cttggagcga ctgtcgttag ttccgcgcga ttcggttcgc
    13201 tcaaatggtt ccgagtggtt catttcgtct caatagaaat tagtaataaa tatttgtatg
    13261 tacaatttat ttgctccaat atatttgtat atatttccct cacagctata tttattctaa
    13321 tttaatatta tgacttttta aggtaatttt ttgtgacctg ttcggagtga ttagcgttac
    13381 aatttgaact gaaagtgaca tccagtgttt gttccttgtg tagatgcatc tcaaaaaaat
    13441 ggtgggcata atagtgttgt ttatatatat caaaaataac aactataata ataagaatac
    13501 atttaattta gaaaatgctt ggatttcact ggaactaggc tagcataact tcgtataatg
    13561 tatgctatac gaagttatgc tagcggatcc aagcttgcgg ccgcaattct atattctaaa
    13621 aacacaaatg atacttctaa aaaaaaatca tgaatggcat caactctgaa tcaaatcttt
    13681 gcagatgcac ctacttctca tttccactgt cacatcattt ttccagatct cgctgcctgt
    13741 tatgtggccc acaaaccaag acacgtttta tggccattaa agctggctga tcgtcgccaa
    13801 acaccaaata cataatgaat atgtacacat tcgagaaaga agcgatcaaa gaagcgtctt
    13861 cgggcggagt aggagaatgc ggaggagaag gagaacgagc tgatctagta tctctccaca
    13921 atccaatgcc aactgaccaa ctggccatat tcggagcaat ttgaagccaa tttccatcgc
    13981 ctggcgatcg ctccattctt ggctatatgt ttttcaccgt tacccggggc cattttcaaa
    14041 gactcgtcgg caagataaga ttgtgtcact cgctgtctct cttcatttgt cgaagaatgc
    14101 tgaggaattt cgcgatgacg tcggcgagta ttttgaagaa tgagaataat ttgtatttat
    14161 acgaaaatca gttagtggaa ttttctacaa aaacatgtta tctatagata attttgttgc
    14221 aaaatatgtt gactatgaca aagattgtat gtatatacct ttaatgtatt ctcattttct
    14281 tatgtattta taatggcaat gatgatactg atgatatttt aagatgatgc cagaccaaaa
    14341 ggcttgaatt tctgcgtctt ttgccgaacg cagtgcatgt gcaattgttg ttttttggaa
    14401 tattcaattt tcggactgtc cgctttgatt tcagtttctt ggcttattca aaaagcaaag
    14461 taaagccaaa aaagcgagat ggcaatacca aatgcggcaa aacggtagtg gaaggaaagg
    14521 ggtgcggggc agcggaagga agggtggggc ggggcgtggc ggggtctgtg gctgggcgcg
    14581 acgtcaccga cgttggagcc actcctttga ccatgtgtgc gtgtgtgtat tattcgtgtc
    14641 tcgccactcg ccggttgttt ttttcttttt atgctgcgct ctctctagcg ccatctcgct
    14701 tacgcatgct caacgcaccg catgttgccg tttcctttta tgcgtcattt tggctcgaaa
    14761 taggcaatta tttaaacaaa gattagtcaa cgaaaacgct aaaataaata agtctacaat
    14821 atggttactt attgccatgt gtgtgcagcc aacgatagca acaaaagcaa caacacaggt
    14881 ggctttccct ctttcacttt ttgtttgcaa gccgcgtgcg agcaagacgg cacgaccggc
    14941 aaacgcaatt acgctgacaa agagcagacg aagttttggc gaaaaacatc aaggcgcctg
    15001 atacgaatgc atttgcaata acaattgcga tatttaatat tgtttatgaa gctgtttgac
    15061 ttcaaaacac acaaaaaaaa aaaataaaac aaattatttg aaagagaatt aggaatcgga
    15121 cgcttatcgt tagggtaaca acaagaaatg cttactgagt cacagcctct ggaaaactgc
    15181 cgcaagccag agagagagag aaaaagaggg agagcagctt agaccgcatg tgcttgtgtg
    15241 tgaggcgtct ctctcttcgt ctctgttgcg caaacgcata gactgcactg aaaaaatcga
    15301 ttacctattt tttatgaatg aatatttgca ctattactat tcaaaactat taagatagca
    15361 atcacattca atagccaaat actataccac ctgagcgatg caacgaaatg atcaatttga
    15421 gcaaaaatgc tgcatattta ggacggcatc attatagaaa tgcttcttgc tgtgtacttt
    15481 tctctcgtct ggcagctgtt tcgccgttat tgttaaaacc ggcttaagtt aggtgtgttt
    15541 tctacgacta gtgaatgccc tactagaaga tgtgtgttgc acaaaatgtc cctggaataa
    15601 ccaatttgaa gtgcagatag cagtaaacgt aagctaatat gaatattatt taactgtaat
    15661 gttttaatat cgctggacat tactaataaa cccactataa acacatgtac atatgtatgt
    15721 tttggcatac aatgagtagt tggggaaaaa atgtgtaaaa gcaccgtgac catcacagca
    15781 taaagataac cagctgaagt atcgaatatg agtaaccccc aaattgaatc acatgccgca
    15841 actgatagga cccatggaag tacactcttc atggcgatat acaagacaca cacaagcacg
    15901 aacacccagt tgcggaggaa attctccgta aatgaaaacc caatcggcga acaattcata
    15961 cccatatatg gtaaaagttt tgaacgcgac ttgagagcgg agagcattgc ggctgataag
    16021 gttttagcgc taagcgggct ttataaaacg ggctgcggga ccagttttca tatcactacc
    16081 gtttgagttc ttgtgctgtg tggatactcc tcccgacaca aagccgctcc atcagccagc
    16141 agtcgtctaa tccagagaca ccaaaccgaa agacttaatt tatatttatt taattaattt
    16201 taataaaaca caccaaatgt aagtagcttt ccccttccca acaacaaaac accatcgaac
    16261 cactcccacc aagaaaaagc aataatcgag aaaagccgcg gaaaatgtgt gatttttttt
    16321 gtaaacaaaa cattttttta tgtgccagtg ctgaaagtga tcaaaaaata ctagccacga
    16381 gctaaagagt tattgtattg accaaaactc caaaaatacc caagtttggc cctaaattgt
    16441 caatcaaaat accaataggt cgaaagacat caaaattaac aaaaccaggg tttcaaatac
    16501 cataactcaa gaatcaggat tacaactgca gatttcagga tatatacata caaattatac
    16561 gaaattataa aaaccaaagc aattcaatag ccccaactca aatgttagga tctaatatag
    16621 tgtttaaagc caagctcgct gatgtgggcg tgtcacgatt tcacccaaag atatgccaaa
    16681 ttacgaattg caaatcaatt cgccaacact ttcttttttt cccacgccct aaaacaccag
    16741 atcatccata aatgtacata catacagtat atgcatatta taatctgtaa aactagatca
    16801 ggttcttgaa aatagtgacg taggcagccg ttttggctga agcagaaatt tttgccggtt
    16861 tttcaaagtt gtagttgcaa aaatggagaa aaccttcgag cattcgttca tatacacaca
    16921 ctcacgcgca aaataacgag agagagtgta tgtgtgtgtg agagagcgaa agccagacga
    16981 cggtttgctt ttcgcctcga aacatgacca tatatggtca caaaacttgg ccgccgcaat
    17041 tcaacacacc agcgctctcc ttcgcaccca tagcgaccat ggcgcggagc gagcgagatg
    17101 gcgagagcga gcgacgccta tggcgacgtc gacgcaggca gcgattgaaa aacgcagtta
    17161 actggcattc aacattcacc agccactttc agtcggttta ttccagtcat tcctttcaaa
    17221 ccgtgcggtc gcttagctca gcctcgccac ttgcgtttac agtagttttc acgccttgaa
    17281 tttgttaaat cgaacaaaaa ggtaaagttt aactagcttt gaaaagtttc gtggctctta
    17341 attgttaaat tttctagagt gcgtttagtg tttttttttt tttttatttt gtaatgttaa
    17401 tttcgggttc caattcgagt tttaggcagc cgccatttta agggcgcata cacacaggca
    17461 actgtgctct ctttgcggct ttcttttgca ccggcattcg ttaagtgtcg tctagaagct
    17521 tctcccctcc cttttcggca tattcgtatt gtggttttaa tttttcgggg cggggccttc
    17581 tattttgtaa ctgttctttt aatttcttat tacaattcga tcgcaagtga aaatcagttt
    17641 tcaatcggaa aagtattttt ttatgaaatt tttttttgtc
//
