LOCUS       sequence-314827-       11061 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 11061)
  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 11061)
  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: ABE7.10-F148A
FEATURES             Location/Qualifiers
     source          1..11061
                     /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        447..826
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        827..1030
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1075..1094
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1075..1093
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2316..5714,5715..5717,5718..6314,6315,6316..6318,
                     6319..6320,6321..6323,6324..6416)
                     /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(2316..2339,2340..2342,2343..6416)
                     /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             6429..6449
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6453..6660
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6729..7032
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7033..7236
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(7265..7267,7268..7270,7271..7984)
                     /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(7265..7267,7268..7270,7271..7981)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7310..7331)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7571..7590)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7918..7939
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      8539..8994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8626..8645)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8836..8857
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9011..9030)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9130..9152
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9190..9208)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9276..9380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9381..9449,9450..10241)
                     /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(9599..9618)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10412..11000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10901..10920
                     /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 aacacaaagc atagactgcg ttttagagct agaaatagca agttaaaata
      301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag
      361 ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag
      421 acaaatggct ctagctcgag cgcgttgaca ttgattattg actagttatt aatagtaatc
      481 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      541 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      601 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      661 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      721 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      781 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      841 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      901 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      961 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     1021 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     1081 gactcactat agggagaccc aagctggcta gcgccaccat gagcgaggtg gagttcagcc
     1141 acgagtactg gatgagacac gcgctgaccc tggcgaagag agcttgggat gagagagagg
     1201 tgccggtggg agctgtgttg gtgcataaca acagagtgat aggagaggga tggaacagac
     1261 ccattgggag gcacgaccca accgctcatg ctgaaataat ggctttgaga cagggagggc
     1321 tggtgatgca gaattacaga ctgatagacg ccactctgta tgtgacactg gagccctgcg
     1381 tgatgtgcgc aggagctatg attcactcca ggatcggcag agtggtgttc ggagctagag
     1441 atgccaaaac cggcgcagcc ggaagcctta tggacgttct tcaccacccc ggcatgaatc
     1501 accgtgtgga aattaccgag ggcatactgg ccgacgagtg cgctgctctt cttagcgact
     1561 tcgctagaat gagacgacag gagatcaagg ctcagaaaaa ggcccaaagc agcaccgaca
     1621 gcggcggatc tagcggagga tctagcggta gcgagacccc tggaacctcc gaaagcgcta
     1681 ccccagagag cagcggagga tcatcaggag gaagcagcga agtagaattt agccacgaat
     1741 attggatgag acatgcactg acactggcca agagagctag agacgagaga gaggttcccg
     1801 taggagccgt gctggtgctt aacaacagag tcataggaga ggggtggaac agagccatcg
     1861 ggcttcacga ccctacagcc catgcagaga taatggcact gagacaggga ggtttggtga
     1921 tgcagaacta taggctgata gacgcaacac tgtacgtgac attcgagcca tgtgtgatgt
     1981 gcgccggagc catgatacac agtagaatcg ggagagtggt gtttggagtg aggaatgcca
     2041 agacaggagc agccggcagc ctgatggacg ttttgcacta ccctggcatg aaccacaggg
     2101 tggagatcac cgagggaatc ttggccgatg agtgcgccgc tctgctgtgt tactttgcta
     2161 ggatgcccag acaagtgttc aacgcccaaa agaaagccca atctagcaca gacagcggcg
     2221 ggtctagcgg aggctcttct ggaagtgaga ctcccggaac ttccgagagc gccacaccag
     2281 aatcaagcgg cggatcgagc ggaggaagcg gatccgataa aaagtattct attggtttag
     2341 ccatcggcac taattccgtt ggatgggctg tcataaccga tgaatacaaa gtaccttcaa
     2401 agaaatttaa ggtgttgggg aacacagacc gtcattcgat taaaaagaat cttatcggtg
     2461 ccctcctatt cgatagtggc gaaacggcag aggcgactcg cctgaaacga accgctcgga
     2521 gaaggtatac acgtcgcaag aaccgaatat gttacttaca agaaattttt agcaatgaga
     2581 tggccaaagt tgacgattct ttctttcacc gtttggaaga gtccttcctt gtcgaagagg
     2641 acaagaaaca tgaacggcac cccatctttg gaaacatagt agatgaggtg gcatatcatg
     2701 aaaagtaccc aacgatttat cacctcagaa aaaagctagt tgactcaact gataaagcgg
     2761 acctgaggtt aatctacttg gctcttgccc atatgataaa gttccgtggg cactttctca
     2821 ttgagggtga tctaaatccg gacaactcgg atgtcgacaa actgttcatc cagttagtac
     2881 aaacctataa tcagttgttt gaagagaacc ctataaatgc aagtggcgtg gatgcgaagg
     2941 ctattcttag cgcccgcctc tctaaatccc gacggctaga aaacctgatc gcacaattac
     3001 ccggagagaa gaaaaatggg ttgttcggta accttatagc gctctcacta ggcctgacac
     3061 caaattttaa gtcgaacttc gacttagctg aagatgccaa attgcagctt agtaaggaca
     3121 cgtacgatga cgatctcgac aatctactgg cacaaattgg agatcagtat gcggacttat
     3181 ttttggctgc caaaaacctt agcgatgcaa tcctcctatc tgacatactg agagttaata
     3241 ctgagattac caaggcgccg ttatccgctt caatgatcaa aaggtacgat gaacatcacc
     3301 aagacttgac acttctcaag gccctagtcc gtcagcaact gcctgagaaa tataaggaaa
     3361 tattctttga tcagtcgaaa aacgggtacg caggttatat tgacggcgga gcgagtcaag
     3421 aggaattcta caagtttatc aaacccatat tagagaagat ggatgggacg gaagagttgc
     3481 ttgtaaaact caatcgcgaa gatctactgc gaaagcagcg gactttcgac aacggtagca
     3541 ttccacatca aatccactta ggcgaattgc atgctatact tagaaggcag gaggattttt
     3601 atccgttcct caaagacaat cgtgaaaaga ttgagaaaat cctaaccttt cgcatacctt
     3661 actatgtggg acccctggcc cgagggaact ctcggttcgc atggatgaca agaaagtccg
     3721 aagaaacgat tactccatgg aattttgagg aagttgtcga taaaggtgcg tcagctcaat
     3781 cgttcatcga gaggatgacc aactttgaca agaatttacc gaacgaaaaa gtattgccta
     3841 agcacagttt actttacgag tatttcacag tgtacaatga actcacgaaa gttaagtatg
     3901 tcactgaggg catgcgtaaa cccgcctttc taagcggaga acagaagaaa gcaatagtag
     3961 atctgttatt caagaccaac cgcaaagtga cagttaagca attgaaagag gactacttta
     4021 agaaaattga atgcttcgat tctgtcgaga tctccggggt agaagatcga tttaatgcgt
     4081 cacttggtac gtatcatgac ctcctaaaga taattaaaga taaggacttc ctggataacg
     4141 aagagaatga agatatctta gaagatatag tgttgactct taccctcttt gaagatcggg
     4201 aaatgattga ggaaagacta aaaacatacg ctcacctgtt cgacgataag gttatgaaac
     4261 agttaaagag gcgtcgctat acgggctggg gacgattgtc gcggaaactt atcaacggga
     4321 taagagacaa gcaaagtggt aaaactattc tcgattttct aaagagcgac ggcttcgcca
     4381 ataggaactt tatgcagctg atccatgatg actctttaac cttcaaagag gatatacaaa
     4441 aggcacaggt ttccggacaa ggggactcat tgcacgaaca tattgcgaat cttgctggtt
     4501 cgccagccat caaaaagggc atactccaga cagtcaaagt agtggatgag ctagttaagg
     4561 tcatgggacg tcacaaaccg gaaaacattg taatcgagat ggcacgcgaa aatcaaacga
     4621 ctcagaaggg gcaaaaaaac agtcgagagc ggatgaagag aatagaagag ggtattaaag
     4681 aactgggcag ccagatctta aaggagcatc ctgtggaaaa tacccaattg cagaacgaga
     4741 aactttacct ctattaccta caaaatggaa gggacatgta tgttgatcag gaactggaca
     4801 taaaccgttt atctgattac gacgtcgatc acattgtacc ccaatccttt ttgaaggacg
     4861 attcaatcga caataaagtg cttacacgct cggataagaa ccgagggaaa agtgacaatg
     4921 ttccaagcga ggaagtcgta aagaaaatga agaactattg gcggcagctc ctaaatgcga
     4981 aactgataac gcaaagaaag ttcgataact taactaaagc tgagaggggt ggcttgtctg
     5041 aacttgacaa ggccggattt attaaacgtc agctcgtgga aacccgccaa atcacaaagc
     5101 atgttgcaca gatactagat tcccgaatga atacgaaata cgacgagaac gataagctga
     5161 ttcgggaagt caaagtaatc actttaaagt caaaattggt gtcggacttc agaaaggatt
     5221 ttcaattcta taaagttagg gagataaata actaccacca tgcgcacgac gcttatctta
     5281 atgccgtcgt agggaccgca ctcattaaga aatacccgaa gctagaaagt gagtttgtgt
     5341 atggtgatta caaagtttat gacgtccgta agatgatcgc gaaaagcgaa caggagatag
     5401 gcaaggctac agccaaatac ttcttttatt ctaacattat gaatttcttt aagacggaaa
     5461 tcactctggc aaacggagag atacgcaaac gacctttaat tgaaaccaat ggggagacag
     5521 gtgaaatcgt atgggataag ggccgggact tcgcgacggt gagaaaagtt ttgtccatgc
     5581 cccaagtcaa catagtaaag aaaactgagg tgcagaccgg agggttttca aaggaatcga
     5641 ttcttccaaa aaggaatagt gataagctca tcgctcgtaa aaaggactgg gacccgaaaa
     5701 agtacggtgg cttcgatagc cctacagttg cctattctgt cctagtagtg gcaaaagttg
     5761 agaagggaaa atccaagaaa ctgaagtcag tcaaagaatt attggggata acgattatgg
     5821 agcgctcgtc ttttgaaaag aaccccatcg acttccttga ggcgaaaggt tacaaggaag
     5881 taaaaaagga tctcataatt aaactaccaa agtatagtct gtttgagtta gaaaatggcc
     5941 gaaaacggat gttggctagc gccggagagc ttcaaaaggg gaacgaactc gcactaccgt
     6001 ctaaatacgt gaatttcctg tatttagcgt cccattacga gaagttgaaa ggttcacctg
     6061 aagataacga acagaagcaa ctttttgttg agcagcacaa acattatctc gacgaaatca
     6121 tagagcaaat ttcggaattc agtaagagag tcatcctagc tgatgccaat ctggacaaag
     6181 tattaagcgc atacaacaag cacagggata aacccatacg tgagcaggcg gaaaatatta
     6241 tccatttgtt tactcttacc aacctcggcg ctccagccgc attcaagtat tttgacacaa
     6301 cgatagatcg caaacgatac acttctacca aggaggtgct agacgcgaca ctgattcacc
     6361 aatccatcac gggattatat gaaactcgga tagatttgtc acagcttggg ggtgactctg
     6421 gtggttctcc caagaagaag aggaaagtct aactgtgcct tctagttgcc agccatctgt
     6481 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     6541 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     6601 tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga
     6661 gcggccgcta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     6721 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     6781 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6841 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6901 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6961 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     7021 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     7081 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     7141 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     7201 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
     7261 tagcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     7321 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     7381 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     7441 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     7501 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     7561 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     7621 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     7681 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     7741 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     7801 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     7861 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     7921 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     7981 gtaaggatcc gggatgcaga aattgatgat ctattaaaca ataaagatgt ccactaaaat
     8041 ggaagttttt cctgtcatac tttgttaaga agggtgagaa cagagtacct acattttgaa
     8101 tggaaggatt ggagctacgg gggtgggggt ggggtgggat tagataaatg cctgctcttt
     8161 actgaaggct ctttactatt gctttatgat aatgtttcat agttggatat cataatttaa
     8221 acaagcaaaa ccaaattaag ggccagctca ttcctcccac tcatgatcta tagatctata
     8281 gatctctcgt gggatcattg tttttctctt gattcccact ttgtggttct aagtactgtg
     8341 gtttccaaat gtgtcagttt catagcctga agaacgagat cagcagcctc tgttccacat
     8401 acacttcatt ctcagtattg ttttgccaag ttctaattcc atcagaagct ggtcgacctg
     8461 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     8521 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     8581 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     8641 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     8701 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     8761 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     8821 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     8881 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     8941 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     9001 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     9061 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     9121 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     9181 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     9241 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     9301 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     9361 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     9421 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     9481 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     9541 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     9601 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     9661 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     9721 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     9781 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     9841 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     9901 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     9961 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
    10021 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
    10081 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct
    10141 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
    10201 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
    10261 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
    10321 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
    10381 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
    10441 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
    10501 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
    10561 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
    10621 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
    10681 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
    10741 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
    10801 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
    10861 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
    10921 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
    10981 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
    11041 gctggccttt tgctcacatg t
//
