LOCUS       sequence-314816-       11047 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 11047)
  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 11047)
  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-YE1
FEATURES             Location/Qualifiers
     source          1..11047
                     /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"
     CDS             join(1961..5359,5360..5362,5363..5959,5960,5961..5963,
                     5964..5965,5966..5968,5969..6061)
                     /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(1961..1984,1985..1987,1988..6061)
                     /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             6074..6322
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6335..6355
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6364..6381
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6433..6450)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6439..6646
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6715..7018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7019..7222
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7173..7193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(7251..7253,7254..7256,7257..7970)
                     /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(7251..7253,7254..7256,7257..7967)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7296..7317)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7557..7576)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7904..7925
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      8525..8980
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8612..8631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8822..8843
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8997..9016)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9116..9138
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9176..9194)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9262..9366
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9367..9435,9436..10227)
                     /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(9585..9604)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10398..10986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10887..10906
                     /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 ccaccatgag ctcagagact ggcccagtgg ctgtggaccc
     1261 cacattgaga cggcggatcg agccccatga gtttgaggta ttcttcgatc cgagagagct
     1321 ccgcaaggag acctgcctgc tttacgaaat taattggggg ggccggcact ccatttggcg
     1381 acatacatca cagaacacta acaagcacgt cgaagtcaac ttcatcgaga agttcacgac
     1441 agaaagatat ttctgtccga acacaaggtg cagcattacc tggtttctca gctacagccc
     1501 atgcggcgaa tgtagtaggg ccatcactga attcctgtca aggtatcccc acgtcactct
     1561 gtttatttac atcgcaaggc tgtaccacca cgctgacccc gagaatcgac aaggcctgcg
     1621 ggatttgatc tcttcaggtg tgactatcca aattatgact gagcaggagt caggatactg
     1681 ctggagaaac tttgtgaatt atagcccgag taatgaagcc cactggccta ggtatcccca
     1741 tctgtgggta cgactgtacg ttcttgaact gtactgcatc atactgggcc tgcctccttg
     1801 tctcaacatt ctgagaagga agcagccaca gctgacattc tttaccatcg ctcttcagtc
     1861 ttgtcattac cagcgactgc ccccacacat tctctgggcc accgggttga aaagcggcag
     1921 cgagactccc gggacctcag agtccgccac acccgaaagt gataaaaagt attctattgg
     1981 tttagccatc ggcactaatt ccgttggatg ggctgtcata accgatgaat acaaagtacc
     2041 ttcaaagaaa tttaaggtgt tggggaacac agaccgtcat tcgattaaaa agaatcttat
     2101 cggtgccctc ctattcgata gtggcgaaac ggcagaggcg actcgcctga aacgaaccgc
     2161 tcggagaagg tatacacgtc gcaagaaccg aatatgttac ttacaagaaa tttttagcaa
     2221 tgagatggcc aaagttgacg attctttctt tcaccgtttg gaagagtcct tccttgtcga
     2281 agaggacaag aaacatgaac ggcaccccat ctttggaaac atagtagatg aggtggcata
     2341 tcatgaaaag tacccaacga tttatcacct cagaaaaaag ctagttgact caactgataa
     2401 agcggacctg aggttaatct acttggctct tgcccatatg ataaagttcc gtgggcactt
     2461 tctcattgag ggtgatctaa atccggacaa ctcggatgtc gacaaactgt tcatccagtt
     2521 agtacaaacc tataatcagt tgtttgaaga gaaccctata aatgcaagtg gcgtggatgc
     2581 gaaggctatt cttagcgccc gcctctctaa atcccgacgg ctagaaaacc tgatcgcaca
     2641 attacccgga gagaagaaaa atgggttgtt cggtaacctt atagcgctct cactaggcct
     2701 gacaccaaat tttaagtcga acttcgactt agctgaagat gccaaattgc agcttagtaa
     2761 ggacacgtac gatgacgatc tcgacaatct actggcacaa attggagatc agtatgcgga
     2821 cttatttttg gctgccaaaa accttagcga tgcaatcctc ctatctgaca tactgagagt
     2881 taatactgag attaccaagg cgccgttatc cgcttcaatg atcaaaaggt acgatgaaca
     2941 tcaccaagac ttgacacttc tcaaggccct agtccgtcag caactgcctg agaaatataa
     3001 ggaaatattc tttgatcagt cgaaaaacgg gtacgcaggt tatattgacg gcggagcgag
     3061 tcaagaggaa ttctacaagt ttatcaaacc catattagag aagatggatg ggacggaaga
     3121 gttgcttgta aaactcaatc gcgaagatct actgcgaaag cagcggactt tcgacaacgg
     3181 tagcattcca catcaaatcc acttaggcga attgcatgct atacttagaa ggcaggagga
     3241 tttttatccg ttcctcaaag acaatcgtga aaagattgag aaaatcctaa cctttcgcat
     3301 accttactat gtgggacccc tggcccgagg gaactctcgg ttcgcatgga tgacaagaaa
     3361 gtccgaagaa acgattactc catggaattt tgaggaagtt gtcgataaag gtgcgtcagc
     3421 tcaatcgttc atcgagagga tgaccaactt tgacaagaat ttaccgaacg aaaaagtatt
     3481 gcctaagcac agtttacttt acgagtattt cacagtgtac aatgaactca cgaaagttaa
     3541 gtatgtcact gagggcatgc gtaaacccgc ctttctaagc ggagaacaga agaaagcaat
     3601 agtagatctg ttattcaaga ccaaccgcaa agtgacagtt aagcaattga aagaggacta
     3661 ctttaagaaa attgaatgct tcgattctgt cgagatctcc ggggtagaag atcgatttaa
     3721 tgcgtcactt ggtacgtatc atgacctcct aaagataatt aaagataagg acttcctgga
     3781 taacgaagag aatgaagata tcttagaaga tatagtgttg actcttaccc tctttgaaga
     3841 tcgggaaatg attgaggaaa gactaaaaac atacgctcac ctgttcgacg ataaggttat
     3901 gaaacagtta aagaggcgtc gctatacggg ctggggacga ttgtcgcgga aacttatcaa
     3961 cgggataaga gacaagcaaa gtggtaaaac tattctcgat tttctaaaga gcgacggctt
     4021 cgccaatagg aactttatgc agctgatcca tgatgactct ttaaccttca aagaggatat
     4081 acaaaaggca caggtttccg gacaagggga ctcattgcac gaacatattg cgaatcttgc
     4141 tggttcgcca gccatcaaaa agggcatact ccagacagtc aaagtagtgg atgagctagt
     4201 taaggtcatg ggacgtcaca aaccggaaaa cattgtaatc gagatggcac gcgaaaatca
     4261 aacgactcag aaggggcaaa aaaacagtcg agagcggatg aagagaatag aagagggtat
     4321 taaagaactg ggcagccaga tcttaaagga gcatcctgtg gaaaataccc aattgcagaa
     4381 cgagaaactt tacctctatt acctacaaaa tggaagggac atgtatgttg atcaggaact
     4441 ggacataaac cgtttatctg attacgacgt cgatcacatt gtaccccaat cctttttgaa
     4501 ggacgattca atcgacaata aagtgcttac acgctcggat aagaaccgag ggaaaagtga
     4561 caatgttcca agcgaggaag tcgtaaagaa aatgaagaac tattggcggc agctcctaaa
     4621 tgcgaaactg ataacgcaaa gaaagttcga taacttaact aaagctgaga ggggtggctt
     4681 gtctgaactt gacaaggccg gatttattaa acgtcagctc gtggaaaccc gccaaatcac
     4741 aaagcatgtt gcacagatac tagattcccg aatgaatacg aaatacgacg agaacgataa
     4801 gctgattcgg gaagtcaaag taatcacttt aaagtcaaaa ttggtgtcgg acttcagaaa
     4861 ggattttcaa ttctataaag ttagggagat aaataactac caccatgcgc acgacgctta
     4921 tcttaatgcc gtcgtaggga ccgcactcat taagaaatac ccgaagctag aaagtgagtt
     4981 tgtgtatggt gattacaaag tttatgacgt ccgtaagatg atcgcgaaaa gcgaacagga
     5041 gataggcaag gctacagcca aatacttctt ttattctaac attatgaatt tctttaagac
     5101 ggaaatcact ctggcaaacg gagagatacg caaacgacct ttaattgaaa ccaatgggga
     5161 gacaggtgaa atcgtatggg ataagggccg ggacttcgcg acggtgagaa aagttttgtc
     5221 catgccccaa gtcaacatag taaagaaaac tgaggtgcag accggagggt tttcaaagga
     5281 atcgattctt ccaaaaagga atagtgataa gctcatcgct cgtaaaaagg actgggaccc
     5341 gaaaaagtac ggtggcttcg atagccctac agttgcctat tctgtcctag tagtggcaaa
     5401 agttgagaag ggaaaatcca agaaactgaa gtcagtcaaa gaattattgg ggataacgat
     5461 tatggagcgc tcgtcttttg aaaagaaccc catcgacttc cttgaggcga aaggttacaa
     5521 ggaagtaaaa aaggatctca taattaaact accaaagtat agtctgtttg agttagaaaa
     5581 tggccgaaaa cggatgttgg ctagcgccgg agagcttcaa aaggggaacg aactcgcact
     5641 accgtctaaa tacgtgaatt tcctgtattt agcgtcccat tacgagaagt tgaaaggttc
     5701 acctgaagat aacgaacaga agcaactttt tgttgagcag cacaaacatt atctcgacga
     5761 aatcatagag caaatttcgg aattcagtaa gagagtcatc ctagctgatg ccaatctgga
     5821 caaagtatta agcgcataca acaagcacag ggataaaccc atacgtgagc aggcggaaaa
     5881 tattatccat ttgtttactc ttaccaacct cggcgctcca gccgcattca agtattttga
     5941 cacaacgata gatcgcaaac gatacacttc taccaaggag gtgctagacg cgacactgat
     6001 tcaccaatcc atcacgggat tatatgaaac tcggatagat ttgtcacagc ttgggggtga
     6061 ctctggtggt tctactaatc tgtcagatat tattgaaaag gagaccggta agcaactggt
     6121 tatccaggaa tccatcctca tgctcccaga ggaggtggaa gaagtcattg ggaacaagcc
     6181 ggaaagcgat atactcgtgc acaccgccta cgacgagagc accgacgaga atgtcatgct
     6241 tctgactagc gacgcccctg aatacaagcc ttgggctctg gtcatacagg atagcaacgg
     6301 tgagaacaag attaagatgc tctctggtgg ttctcccaag aagaagagga aagtctaacc
     6361 ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctaaga attcctagag
     6421 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6481 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6541 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     6601 acagcaaggg ggaggattgg gaagagaata gcaggcatgc tggggagcgg ccgctagtta
     6661 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     6721 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     6781 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     6841 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     6901 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     6961 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     7021 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     7081 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     7141 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     7201 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc atggtgagca
     7261 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     7321 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     7381 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     7441 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     7501 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     7561 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     7621 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     7681 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     7741 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     7801 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     7861 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     7921 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa ggatccggga
     7981 tgcagaaatt gatgatctat taaacaataa agatgtccac taaaatggaa gtttttcctg
     8041 tcatactttg ttaagaaggg tgagaacaga gtacctacat tttgaatgga aggattggag
     8101 ctacgggggt gggggtgggg tgggattaga taaatgcctg ctctttactg aaggctcttt
     8161 actattgctt tatgataatg tttcatagtt ggatatcata atttaaacaa gcaaaaccaa
     8221 attaagggcc agctcattcc tcccactcat gatctataga tctatagatc tctcgtggga
     8281 tcattgtttt tctcttgatt cccactttgt ggttctaagt actgtggttt ccaaatgtgt
     8341 cagtttcata gcctgaagaa cgagatcagc agcctctgtt ccacatacac ttcattctca
     8401 gtattgtttt gccaagttct aattccatca gaagctggtc gacctgcagg ggcgcctgat
     8461 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     8521 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     8581 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     8641 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     8701 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     8761 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     8821 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     8881 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     8941 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     9001 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     9061 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     9121 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     9181 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     9241 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     9301 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     9361 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     9421 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     9481 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     9541 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     9601 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     9661 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     9721 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     9781 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     9841 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     9901 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     9961 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
    10021 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
    10081 ggaagccgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
    10141 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
    10201 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
    10261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
    10321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
    10381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
    10441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
    10501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
    10561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
    10621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
    10681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
    10741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
    10801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
    10861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
    10921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
    10981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
    11041 cacatgt
//
