LOCUS       sequence-314824-       10957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gene editing. See Gene/Insert section of the plasmid page for 
            targeting sequence cloned into this plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10957)
  AUTHORS   Zhou C, Sun Y, Yan R, Liu Y, Zuo E, Gu C, Han L, Wei Y, Hu X, Zeng 
            R, Li Y, Zhou H, Guo F, Yang H
  TITLE     Off-target RNA mutation induced by DNA base editing and its 
            elimination by mutagenesis.
  JOURNAL   Nature. 2019 Jul;571(7764):275-278. doi: 10.1038/s41586-019-1314-0. 
            Epub 2019 Jun 10.
  PUBMED    31181567
REFERENCE   2  (bases 1 to 10957)
  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: 
            "10.1038/s41586-019-1314-0"; journalName: "Nature"; date: "2019-07";
            volume: "571"; issue: "7764"; pages: "275-278"
COMMENT     Sequence Label: BE3-hA3A-R128A
FEATURES             Location/Qualifiers
     source          1..10957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        270..345
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        442..727
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          952..1179
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     primer_bind     1194..1213
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1194..1212
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1220..1229
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1226..1822
                     /codon_start=1
                     /gene="human APOBEC3A"
                     /gene_synonym="A3A"
                     /product="human cytidine deaminase (C to U editing enzyme) 
                     that efficiently edits methylated cytosines (Wang et al., 
                     2018)"
                     /label=APOBEC3A
                     /note="also known as hA3A"
                     /translation="MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTS
                     VKMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSW
                     GCAGEVRAFLQENTHVRLRIFAAAIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     CDS             join(1871..5269,5270..5272,5273..5869,5870,5871..5873,
                     5874..5875,5876..5878,5879..5971)
                     /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(1871..1894,1895..1897,1898..5971)
                     /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             5984..6232
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6245..6265
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6274..6291
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6343..6360)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6349..6556
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6625..6928
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6929..7132
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7083..7103
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(7161..7163,7164..7166,7167..7880)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7161..7163,7164..7166,7167..7877)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7206..7227)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7467..7486)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7814..7835
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      8435..8890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8522..8541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8732..8753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8907..8926)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9026..9048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9086..9104)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9172..9276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9277..9345,9346..10137)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9495..9514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10308..10896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10797..10816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg gcccagactg agcacgtgag ttttagagct agaaatagca agttaaaata
      301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag
      361 ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag
      421 acaaatggct ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc
      481 gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg
      541 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      601 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca
      661 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      721 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
      781 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcggggcg
      841 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
      901 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
      961 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
     1021 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     1081 aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt
     1141 aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt ggctaatacg
     1201 actcactata gggagagccg ccaccatgga agccagccca gcatccgggc ccagacactt
     1261 gatggatcca cacatattca cttccaactt taacaatggc attggaaggc ataagaccta
     1321 cctgtgctac gaagtggagc gcctggacaa tggcacctcg gtcaagatgg accagcacag
     1381 gggctttcta cacaaccagg ctaagaatct tctctgtggc ttttacggcc gccatgcgga
     1441 gctgcgcttc ttggacctgg ttccttcttt gcagttggac ccggcccaga tctacagggt
     1501 cacttggttc atctcctgga gcccctgctt ctcctggggc tgtgccgggg aagtgcgtgc
     1561 gttccttcag gagaacacac acgtgagact gcgtatcttc gctgccgcca tctatgatta
     1621 cgacccccta tataaggagg cactgcaaat gctgcgggat gctggggccc aagtctccat
     1681 catgacctac gatgaattta agcactgctg ggacaccttt gtggaccacc agggatgtcc
     1741 cttccagccc tgggatggac tagatgagca cagccaagcc ctgagtggga ggctgcgggc
     1801 cattctccag aatcagggaa acagcggcag cgagactccc gggacctcag agtccgccac
     1861 acccgaaagt gataaaaagt attctattgg tttagccatc ggcactaatt ccgttggatg
     1921 ggctgtcata accgatgaat acaaagtacc ttcaaagaaa tttaaggtgt tggggaacac
     1981 agaccgtcat tcgattaaaa agaatcttat cggtgccctc ctattcgata gtggcgaaac
     2041 ggcagaggcg actcgcctga aacgaaccgc tcggagaagg tatacacgtc gcaagaaccg
     2101 aatatgttac ttacaagaaa tttttagcaa tgagatggcc aaagttgacg attctttctt
     2161 tcaccgtttg gaagagtcct tccttgtcga agaggacaag aaacatgaac ggcaccccat
     2221 ctttggaaac atagtagatg aggtggcata tcatgaaaag tacccaacga tttatcacct
     2281 cagaaaaaag ctagttgact caactgataa agcggacctg aggttaatct acttggctct
     2341 tgcccatatg ataaagttcc gtgggcactt tctcattgag ggtgatctaa atccggacaa
     2401 ctcggatgtc gacaaactgt tcatccagtt agtacaaacc tataatcagt tgtttgaaga
     2461 gaaccctata aatgcaagtg gcgtggatgc gaaggctatt cttagcgccc gcctctctaa
     2521 atcccgacgg ctagaaaacc tgatcgcaca attacccgga gagaagaaaa atgggttgtt
     2581 cggtaacctt atagcgctct cactaggcct gacaccaaat tttaagtcga acttcgactt
     2641 agctgaagat gccaaattgc agcttagtaa ggacacgtac gatgacgatc tcgacaatct
     2701 actggcacaa attggagatc agtatgcgga cttatttttg gctgccaaaa accttagcga
     2761 tgcaatcctc ctatctgaca tactgagagt taatactgag attaccaagg cgccgttatc
     2821 cgcttcaatg atcaaaaggt acgatgaaca tcaccaagac ttgacacttc tcaaggccct
     2881 agtccgtcag caactgcctg agaaatataa ggaaatattc tttgatcagt cgaaaaacgg
     2941 gtacgcaggt tatattgacg gcggagcgag tcaagaggaa ttctacaagt ttatcaaacc
     3001 catattagag aagatggatg ggacggaaga gttgcttgta aaactcaatc gcgaagatct
     3061 actgcgaaag cagcggactt tcgacaacgg tagcattcca catcaaatcc acttaggcga
     3121 attgcatgct atacttagaa ggcaggagga tttttatccg ttcctcaaag acaatcgtga
     3181 aaagattgag aaaatcctaa cctttcgcat accttactat gtgggacccc tggcccgagg
     3241 gaactctcgg ttcgcatgga tgacaagaaa gtccgaagaa acgattactc catggaattt
     3301 tgaggaagtt gtcgataaag gtgcgtcagc tcaatcgttc atcgagagga tgaccaactt
     3361 tgacaagaat ttaccgaacg aaaaagtatt gcctaagcac agtttacttt acgagtattt
     3421 cacagtgtac aatgaactca cgaaagttaa gtatgtcact gagggcatgc gtaaacccgc
     3481 ctttctaagc ggagaacaga agaaagcaat agtagatctg ttattcaaga ccaaccgcaa
     3541 agtgacagtt aagcaattga aagaggacta ctttaagaaa attgaatgct tcgattctgt
     3601 cgagatctcc ggggtagaag atcgatttaa tgcgtcactt ggtacgtatc atgacctcct
     3661 aaagataatt aaagataagg acttcctgga taacgaagag aatgaagata tcttagaaga
     3721 tatagtgttg actcttaccc tctttgaaga tcgggaaatg attgaggaaa gactaaaaac
     3781 atacgctcac ctgttcgacg ataaggttat gaaacagtta aagaggcgtc gctatacggg
     3841 ctggggacga ttgtcgcgga aacttatcaa cgggataaga gacaagcaaa gtggtaaaac
     3901 tattctcgat tttctaaaga gcgacggctt cgccaatagg aactttatgc agctgatcca
     3961 tgatgactct ttaaccttca aagaggatat acaaaaggca caggtttccg gacaagggga
     4021 ctcattgcac gaacatattg cgaatcttgc tggttcgcca gccatcaaaa agggcatact
     4081 ccagacagtc aaagtagtgg atgagctagt taaggtcatg ggacgtcaca aaccggaaaa
     4141 cattgtaatc gagatggcac gcgaaaatca aacgactcag aaggggcaaa aaaacagtcg
     4201 agagcggatg aagagaatag aagagggtat taaagaactg ggcagccaga tcttaaagga
     4261 gcatcctgtg gaaaataccc aattgcagaa cgagaaactt tacctctatt acctacaaaa
     4321 tggaagggac atgtatgttg atcaggaact ggacataaac cgtttatctg attacgacgt
     4381 cgatcacatt gtaccccaat cctttttgaa ggacgattca atcgacaata aagtgcttac
     4441 acgctcggat aagaaccgag ggaaaagtga caatgttcca agcgaggaag tcgtaaagaa
     4501 aatgaagaac tattggcggc agctcctaaa tgcgaaactg ataacgcaaa gaaagttcga
     4561 taacttaact aaagctgaga ggggtggctt gtctgaactt gacaaggccg gatttattaa
     4621 acgtcagctc gtggaaaccc gccaaatcac aaagcatgtt gcacagatac tagattcccg
     4681 aatgaatacg aaatacgacg agaacgataa gctgattcgg gaagtcaaag taatcacttt
     4741 aaagtcaaaa ttggtgtcgg acttcagaaa ggattttcaa ttctataaag ttagggagat
     4801 aaataactac caccatgcgc acgacgctta tcttaatgcc gtcgtaggga ccgcactcat
     4861 taagaaatac ccgaagctag aaagtgagtt tgtgtatggt gattacaaag tttatgacgt
     4921 ccgtaagatg atcgcgaaaa gcgaacagga gataggcaag gctacagcca aatacttctt
     4981 ttattctaac attatgaatt tctttaagac ggaaatcact ctggcaaacg gagagatacg
     5041 caaacgacct ttaattgaaa ccaatgggga gacaggtgaa atcgtatggg ataagggccg
     5101 ggacttcgcg acggtgagaa aagttttgtc catgccccaa gtcaacatag taaagaaaac
     5161 tgaggtgcag accggagggt tttcaaagga atcgattctt ccaaaaagga atagtgataa
     5221 gctcatcgct cgtaaaaagg actgggaccc gaaaaagtac ggtggcttcg atagccctac
     5281 agttgcctat tctgtcctag tagtggcaaa agttgagaag ggaaaatcca agaaactgaa
     5341 gtcagtcaaa gaattattgg ggataacgat tatggagcgc tcgtcttttg aaaagaaccc
     5401 catcgacttc cttgaggcga aaggttacaa ggaagtaaaa aaggatctca taattaaact
     5461 accaaagtat agtctgtttg agttagaaaa tggccgaaaa cggatgttgg ctagcgccgg
     5521 agagcttcaa aaggggaacg aactcgcact accgtctaaa tacgtgaatt tcctgtattt
     5581 agcgtcccat tacgagaagt tgaaaggttc acctgaagat aacgaacaga agcaactttt
     5641 tgttgagcag cacaaacatt atctcgacga aatcatagag caaatttcgg aattcagtaa
     5701 gagagtcatc ctagctgatg ccaatctgga caaagtatta agcgcataca acaagcacag
     5761 ggataaaccc atacgtgagc aggcggaaaa tattatccat ttgtttactc ttaccaacct
     5821 cggcgctcca gccgcattca agtattttga cacaacgata gatcgcaaac gatacacttc
     5881 taccaaggag gtgctagacg cgacactgat tcaccaatcc atcacgggat tatatgaaac
     5941 tcggatagat ttgtcacagc ttgggggtga ctctggtggt tctactaatc tgtcagatat
     6001 tattgaaaag gagaccggta agcaactggt tatccaggaa tccatcctca tgctcccaga
     6061 ggaggtggaa gaagtcattg ggaacaagcc ggaaagcgat atactcgtgc acaccgccta
     6121 cgacgagagc accgacgaga atgtcatgct tctgactagc gacgcccctg aatacaagcc
     6181 ttgggctctg gtcatacagg atagcaacgg tgagaacaag attaagatgc tctctggtgg
     6241 ttctcccaag aagaagagga aagtctaacc ggtcatcatc accatcacca ttgagtttaa
     6301 acccgctgat cagcctaaga attcctagag ctcgctgatc agcctcgact gtgccttcta
     6361 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     6421 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     6481 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata
     6541 gcaggcatgc tggggagcgg ccgctagtta ttaatagtaa tcaattacgg ggtcattagt
     6601 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     6661 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     6721 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     6781 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     6841 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     6901 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     6961 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     7021 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     7081 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     7141 gaaccgtcag atccgctagc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     7201 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     7261 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     7321 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     7381 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     7441 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     7501 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     7561 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     7621 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     7681 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     7741 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     7801 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     7861 tggacgagct gtacaagtaa ggatccggga tgcagaaatt gatgatctat taaacaataa
     7921 agatgtccac taaaatggaa gtttttcctg tcatactttg ttaagaaggg tgagaacaga
     7981 gtacctacat tttgaatgga aggattggag ctacgggggt gggggtgggg tgggattaga
     8041 taaatgcctg ctctttactg aaggctcttt actattgctt tatgataatg tttcatagtt
     8101 ggatatcata atttaaacaa gcaaaaccaa attaagggcc agctcattcc tcccactcat
     8161 gatctataga tctatagatc tctcgtggga tcattgtttt tctcttgatt cccactttgt
     8221 ggttctaagt actgtggttt ccaaatgtgt cagtttcata gcctgaagaa cgagatcagc
     8281 agcctctgtt ccacatacac ttcattctca gtattgtttt gccaagttct aattccatca
     8341 gaagctggtc gacctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     8401 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     8461 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     8521 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     8581 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     8641 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     8701 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     8761 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     8821 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     8881 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     8941 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     9001 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     9061 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     9121 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     9181 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     9241 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     9301 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     9361 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     9421 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     9481 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     9541 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     9601 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     9661 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     9721 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     9781 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     9841 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     9901 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     9961 attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg
    10021 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
    10081 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
    10141 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
    10201 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
    10261 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
    10321 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
    10381 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
    10441 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
    10501 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
    10561 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
    10621 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
    10681 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
    10741 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
    10801 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
    10861 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
    10921 cggttcctgg ccttttgctg gccttttgct cacatgt
//
