LOCUS       sequence-314826-       10979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gene editing. See Gene/Insert section of 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 10979)
  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 10979)
  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-Y130F
FEATURES             Location/Qualifiers
     source          1..10979
                     /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          974..1201
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     primer_bind     1216..1235
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1216..1234
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1242..1251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1248..1844
                     /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
                     GCAGEVRAFLQENTHVRLRIFAARIFDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     CDS             join(1893..5291,5292..5294,5295..5891,5892,5893..5895,
                     5896..5897,5898..5900,5901..5993)
                     /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(1893..1916,1917..1919,1920..5993)
                     /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             6006..6254
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6267..6287
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6296..6313
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6365..6382)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6371..6578
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6647..6950
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6951..7154
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7105..7125
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(7183..7185,7186..7188,7189..7902)
                     /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(7183..7185,7186..7188,7189..7899)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7228..7249)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7489..7508)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7836..7857
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      8457..8912
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8544..8563)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8754..8775
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8929..8948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9048..9070
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9108..9126)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9194..9298
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9299..9367,9368..10159)
                     /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(9517..9536)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10330..10918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10819..10838
                     /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 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      901 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      961 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
     1021 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
     1081 cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt
     1141 ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca
     1201 ggttggaccg gtggctaata cgactcacta tagggagagc cgccaccatg gaagccagcc
     1261 cagcatccgg gcccagacac ttgatggatc cacacatatt cacttccaac tttaacaatg
     1321 gcattggaag gcataagacc tacctgtgct acgaagtgga gcgcctggac aatggcacct
     1381 cggtcaagat ggaccagcac aggggctttc tacacaacca ggctaagaat cttctctgtg
     1441 gcttttacgg ccgccatgcg gagctgcgct tcttggacct ggttccttct ttgcagttgg
     1501 acccggccca gatctacagg gtcacttggt tcatctcctg gagcccctgc ttctcctggg
     1561 gctgtgccgg ggaagtgcgt gcgttccttc aggagaacac acacgtgaga ctgcgtatct
     1621 tcgctgcccg catctttgat tacgaccccc tatataagga ggcactgcaa atgctgcggg
     1681 atgctggggc ccaagtctcc atcatgacct acgatgaatt taagcactgc tgggacacct
     1741 ttgtggacca ccagggatgt cccttccagc cctgggatgg actagatgag cacagccaag
     1801 ccctgagtgg gaggctgcgg gccattctcc agaatcaggg aaacagcggc agcgagactc
     1861 ccgggacctc agagtccgcc acacccgaaa gtgataaaaa gtattctatt ggtttagcca
     1921 tcggcactaa ttccgttgga tgggctgtca taaccgatga atacaaagta ccttcaaaga
     1981 aatttaaggt gttggggaac acagaccgtc attcgattaa aaagaatctt atcggtgccc
     2041 tcctattcga tagtggcgaa acggcagagg cgactcgcct gaaacgaacc gctcggagaa
     2101 ggtatacacg tcgcaagaac cgaatatgtt acttacaaga aatttttagc aatgagatgg
     2161 ccaaagttga cgattctttc tttcaccgtt tggaagagtc cttccttgtc gaagaggaca
     2221 agaaacatga acggcacccc atctttggaa acatagtaga tgaggtggca tatcatgaaa
     2281 agtacccaac gatttatcac ctcagaaaaa agctagttga ctcaactgat aaagcggacc
     2341 tgaggttaat ctacttggct cttgcccata tgataaagtt ccgtgggcac tttctcattg
     2401 agggtgatct aaatccggac aactcggatg tcgacaaact gttcatccag ttagtacaaa
     2461 cctataatca gttgtttgaa gagaacccta taaatgcaag tggcgtggat gcgaaggcta
     2521 ttcttagcgc ccgcctctct aaatcccgac ggctagaaaa cctgatcgca caattacccg
     2581 gagagaagaa aaatgggttg ttcggtaacc ttatagcgct ctcactaggc ctgacaccaa
     2641 attttaagtc gaacttcgac ttagctgaag atgccaaatt gcagcttagt aaggacacgt
     2701 acgatgacga tctcgacaat ctactggcac aaattggaga tcagtatgcg gacttatttt
     2761 tggctgccaa aaaccttagc gatgcaatcc tcctatctga catactgaga gttaatactg
     2821 agattaccaa ggcgccgtta tccgcttcaa tgatcaaaag gtacgatgaa catcaccaag
     2881 acttgacact tctcaaggcc ctagtccgtc agcaactgcc tgagaaatat aaggaaatat
     2941 tctttgatca gtcgaaaaac gggtacgcag gttatattga cggcggagcg agtcaagagg
     3001 aattctacaa gtttatcaaa cccatattag agaagatgga tgggacggaa gagttgcttg
     3061 taaaactcaa tcgcgaagat ctactgcgaa agcagcggac tttcgacaac ggtagcattc
     3121 cacatcaaat ccacttaggc gaattgcatg ctatacttag aaggcaggag gatttttatc
     3181 cgttcctcaa agacaatcgt gaaaagattg agaaaatcct aacctttcgc ataccttact
     3241 atgtgggacc cctggcccga gggaactctc ggttcgcatg gatgacaaga aagtccgaag
     3301 aaacgattac tccatggaat tttgaggaag ttgtcgataa aggtgcgtca gctcaatcgt
     3361 tcatcgagag gatgaccaac tttgacaaga atttaccgaa cgaaaaagta ttgcctaagc
     3421 acagtttact ttacgagtat ttcacagtgt acaatgaact cacgaaagtt aagtatgtca
     3481 ctgagggcat gcgtaaaccc gcctttctaa gcggagaaca gaagaaagca atagtagatc
     3541 tgttattcaa gaccaaccgc aaagtgacag ttaagcaatt gaaagaggac tactttaaga
     3601 aaattgaatg cttcgattct gtcgagatct ccggggtaga agatcgattt aatgcgtcac
     3661 ttggtacgta tcatgacctc ctaaagataa ttaaagataa ggacttcctg gataacgaag
     3721 agaatgaaga tatcttagaa gatatagtgt tgactcttac cctctttgaa gatcgggaaa
     3781 tgattgagga aagactaaaa acatacgctc acctgttcga cgataaggtt atgaaacagt
     3841 taaagaggcg tcgctatacg ggctggggac gattgtcgcg gaaacttatc aacgggataa
     3901 gagacaagca aagtggtaaa actattctcg attttctaaa gagcgacggc ttcgccaata
     3961 ggaactttat gcagctgatc catgatgact ctttaacctt caaagaggat atacaaaagg
     4021 cacaggtttc cggacaaggg gactcattgc acgaacatat tgcgaatctt gctggttcgc
     4081 cagccatcaa aaagggcata ctccagacag tcaaagtagt ggatgagcta gttaaggtca
     4141 tgggacgtca caaaccggaa aacattgtaa tcgagatggc acgcgaaaat caaacgactc
     4201 agaaggggca aaaaaacagt cgagagcgga tgaagagaat agaagagggt attaaagaac
     4261 tgggcagcca gatcttaaag gagcatcctg tggaaaatac ccaattgcag aacgagaaac
     4321 tttacctcta ttacctacaa aatggaaggg acatgtatgt tgatcaggaa ctggacataa
     4381 accgtttatc tgattacgac gtcgatcaca ttgtacccca atcctttttg aaggacgatt
     4441 caatcgacaa taaagtgctt acacgctcgg ataagaaccg agggaaaagt gacaatgttc
     4501 caagcgagga agtcgtaaag aaaatgaaga actattggcg gcagctccta aatgcgaaac
     4561 tgataacgca aagaaagttc gataacttaa ctaaagctga gaggggtggc ttgtctgaac
     4621 ttgacaaggc cggatttatt aaacgtcagc tcgtggaaac ccgccaaatc acaaagcatg
     4681 ttgcacagat actagattcc cgaatgaata cgaaatacga cgagaacgat aagctgattc
     4741 gggaagtcaa agtaatcact ttaaagtcaa aattggtgtc ggacttcaga aaggattttc
     4801 aattctataa agttagggag ataaataact accaccatgc gcacgacgct tatcttaatg
     4861 ccgtcgtagg gaccgcactc attaagaaat acccgaagct agaaagtgag tttgtgtatg
     4921 gtgattacaa agtttatgac gtccgtaaga tgatcgcgaa aagcgaacag gagataggca
     4981 aggctacagc caaatacttc ttttattcta acattatgaa tttctttaag acggaaatca
     5041 ctctggcaaa cggagagata cgcaaacgac ctttaattga aaccaatggg gagacaggtg
     5101 aaatcgtatg ggataagggc cgggacttcg cgacggtgag aaaagttttg tccatgcccc
     5161 aagtcaacat agtaaagaaa actgaggtgc agaccggagg gttttcaaag gaatcgattc
     5221 ttccaaaaag gaatagtgat aagctcatcg ctcgtaaaaa ggactgggac ccgaaaaagt
     5281 acggtggctt cgatagccct acagttgcct attctgtcct agtagtggca aaagttgaga
     5341 agggaaaatc caagaaactg aagtcagtca aagaattatt ggggataacg attatggagc
     5401 gctcgtcttt tgaaaagaac cccatcgact tccttgaggc gaaaggttac aaggaagtaa
     5461 aaaaggatct cataattaaa ctaccaaagt atagtctgtt tgagttagaa aatggccgaa
     5521 aacggatgtt ggctagcgcc ggagagcttc aaaaggggaa cgaactcgca ctaccgtcta
     5581 aatacgtgaa tttcctgtat ttagcgtccc attacgagaa gttgaaaggt tcacctgaag
     5641 ataacgaaca gaagcaactt tttgttgagc agcacaaaca ttatctcgac gaaatcatag
     5701 agcaaatttc ggaattcagt aagagagtca tcctagctga tgccaatctg gacaaagtat
     5761 taagcgcata caacaagcac agggataaac ccatacgtga gcaggcggaa aatattatcc
     5821 atttgtttac tcttaccaac ctcggcgctc cagccgcatt caagtatttt gacacaacga
     5881 tagatcgcaa acgatacact tctaccaagg aggtgctaga cgcgacactg attcaccaat
     5941 ccatcacggg attatatgaa actcggatag atttgtcaca gcttgggggt gactctggtg
     6001 gttctactaa tctgtcagat attattgaaa aggagaccgg taagcaactg gttatccagg
     6061 aatccatcct catgctccca gaggaggtgg aagaagtcat tgggaacaag ccggaaagcg
     6121 atatactcgt gcacaccgcc tacgacgaga gcaccgacga gaatgtcatg cttctgacta
     6181 gcgacgcccc tgaatacaag ccttgggctc tggtcataca ggatagcaac ggtgagaaca
     6241 agattaagat gctctctggt ggttctccca agaagaagag gaaagtctaa ccggtcatca
     6301 tcaccatcac cattgagttt aaacccgctg atcagcctaa gaattcctag agctcgctga
     6361 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     6421 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     6481 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     6541 ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgctagt tattaatagt
     6601 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     6661 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     6721 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     6781 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     6841 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     6901 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     6961 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     7021 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     7081 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtcc
     7141 atataagcag agctggttta gtgaaccgtc agatccgcta gcatggtgag caagggcgag
     7201 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     7261 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     7321 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     7381 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     7441 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     7501 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     7561 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     7621 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     7681 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     7741 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     7801 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     7861 gccgccggga tcactctcgg catggacgag ctgtacaagt aaggatccgg gatgcagaaa
     7921 ttgatgatct attaaacaat aaagatgtcc actaaaatgg aagtttttcc tgtcatactt
     7981 tgttaagaag ggtgagaaca gagtacctac attttgaatg gaaggattgg agctacgggg
     8041 gtgggggtgg ggtgggatta gataaatgcc tgctctttac tgaaggctct ttactattgc
     8101 tttatgataa tgtttcatag ttggatatca taatttaaac aagcaaaacc aaattaaggg
     8161 ccagctcatt cctcccactc atgatctata gatctataga tctctcgtgg gatcattgtt
     8221 tttctcttga ttcccacttt gtggttctaa gtactgtggt ttccaaatgt gtcagtttca
     8281 tagcctgaag aacgagatca gcagcctctg ttccacatac acttcattct cagtattgtt
     8341 ttgccaagtt ctaattccat cagaagctgg tcgacctgca ggggcgcctg atgcggtatt
     8401 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     8461 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     8521 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     8581 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     8641 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     8701 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     8761 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     8821 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     8881 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     8941 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     9001 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     9061 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     9121 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     9181 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     9241 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     9301 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     9361 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     9421 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     9481 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     9541 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     9601 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     9661 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     9721 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     9781 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     9841 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     9901 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     9961 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg
    10021 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
    10081 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
    10141 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
    10201 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
    10261 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
    10321 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
    10381 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
    10441 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
    10501 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
    10561 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
    10621 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
    10681 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
    10741 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
    10801 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
    10861 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
    10921 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
