LOCUS       sequence-98221-e        9070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual Expression Vector for FokI-dCas9 and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9070)
  AUTHORS   Miyaoka Y, Berman JR, Cooper SB, Mayerl SJ, Chan AH, Zhang B, 
            Karlin-Neumann GA, Conklin BR
  TITLE     Systematic quantification of HDR and NHEJ reveals effects of locus, 
            nuclease, and cell type on genome-editing.
  JOURNAL   Sci Rep. 2016 Mar 31;6:23549. doi: 10.1038/srep23549.
  PUBMED    27030102
REFERENCE   2  (bases 1 to 9070)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-03-31"; volume: "6:23549. doi"; pages: " 
            10.1038/srep23549"
COMMENT     Sequence Label: pXFokI-dCas9
FEATURES             Location/Qualifiers
     source          1..9070
                     /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        268..343
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        440..725
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        727..1004
                     /label=chicken beta-actin promoter
     intron          1005..1233
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1254..1274
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1281..1868
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     CDS             join(1884..5282,5283..5285,5286..5882,5883,5884..5886,
                     5887..5888,5889..5891,5892..5984)
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1884..1907,1908..1910,1911..4397,4398..4400,
                     4401..5984)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5991..6011
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6018..6083
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6111..6128)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6117..6324
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6333..6473
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6333..6462
                     /label=AAV2 ITR
     rep_origin      6548..7003
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6635..6654)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6845..6866
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7020..7039)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7139..7161
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7199..7217)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7285..7389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7390..7458,7459..8250)
                     /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(7608..7627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8421..9009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8910..8929
                     /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 ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
      301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
      361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
      421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
      541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
      661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
      781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
      841 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
      901 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc
     1021 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     1081 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     1141 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac
     1201 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atgcctaaga
     1261 agaagcggaa ggtgagcagc caacttgtga agtctgaact cgaggagaaa aaatcagagt
     1321 tgagacacaa gttgaagtac gtgccacacg aatacatcga gcttatcgag atcgccagaa
     1381 acagtaccca ggataggatc cttgagatga aagtcatgga gttctttatg aaggtctacg
     1441 gttatagagg aaagcacctt ggcggtagca gaaagcccga tggcgccatc tatactgtcg
     1501 gatctcctat cgattatggg gtgatcgtgg ataccaaagc ttactcaggc gggtacaact
     1561 tgcccatagg acaagccgac gagatgcagc ggtatgtcga agagaaccag acgcgcaaca
     1621 agcacatcaa ccccaatgaa tggtggaaag tgtacccaag tagtgtgact gagttcaagt
     1681 tcctgtttgt ctccggccac tttaagggca attataaagc tcagctcact agactcaatc
     1741 acatcacaaa ctgcaacgga gctgtgttgt cagtggagga gctcctgatt ggaggcgaga
     1801 tgatcaaagc cggcaccctt acactggagg aggtgcggcg gaagttcaac aatggagaga
     1861 tcaacttcgg tggcggtgga tccgataaaa agtattctat tggtttagcc atcggcacta
     1921 attccgttgg atgggctgtc ataaccgatg aatacaaagt accttcaaag aaatttaagg
     1981 tgttggggaa cacagaccgt cattcgatta aaaagaatct tatcggtgcc ctcctattcg
     2041 atagtggcga aacggcagag gcgactcgcc tgaaacgaac cgctcggaga aggtatacac
     2101 gtcgcaagaa ccgaatatgt tacttacaag aaatttttag caatgagatg gccaaagttg
     2161 acgattcttt ctttcaccgt ttggaagagt ccttccttgt cgaagaggac aagaaacatg
     2221 aacggcaccc catctttgga aacatagtag atgaggtggc atatcatgaa aagtacccaa
     2281 cgatttatca cctcagaaaa aagctagttg actcaactga taaagcggac ctgaggttaa
     2341 tctacttggc tcttgcccat atgataaagt tccgtgggca ctttctcatt gagggtgatc
     2401 taaatccgga caactcggat gtcgacaaac tgttcatcca gttagtacaa acctataatc
     2461 agttgtttga agagaaccct ataaatgcaa gtggcgtgga tgcgaaggct attcttagcg
     2521 cccgcctctc taaatcccga cggctagaaa acctgatcgc acaattaccc ggagagaaga
     2581 aaaatgggtt gttcggtaac cttatagcgc tctcactagg cctgacacca aattttaagt
     2641 cgaacttcga cttagctgaa gatgccaaat tgcagcttag taaggacacg tacgatgacg
     2701 atctcgacaa tctactggca caaattggag atcagtatgc ggacttattt ttggctgcca
     2761 aaaaccttag cgatgcaatc ctcctatctg acatactgag agttaatact gagattacca
     2821 aggcgccgtt atccgcttca atgatcaaaa ggtacgatga acatcaccaa gacttgacac
     2881 ttctcaaggc cctagtccgt cagcaactgc ctgagaaata taaggaaata ttctttgatc
     2941 agtcgaaaaa cgggtacgca ggttatattg acggcggagc gagtcaagag gaattctaca
     3001 agtttatcaa acccatatta gagaagatgg atgggacgga agagttgctt gtaaaactca
     3061 atcgcgaaga tctactgcga aagcagcgga ctttcgacaa cggtagcatt ccacatcaaa
     3121 tccacttagg cgaattgcat gctatactta gaaggcagga ggatttttat ccgttcctca
     3181 aagacaatcg tgaaaagatt gagaaaatcc taacctttcg cataccttac tatgtgggac
     3241 ccctggcccg agggaactct cggttcgcat ggatgacaag aaagtccgaa gaaacgatta
     3301 ctccatggaa ttttgaggaa gttgtcgata aaggtgcgtc agctcaatcg ttcatcgaga
     3361 ggatgaccaa ctttgacaag aatttaccga acgaaaaagt attgcctaag cacagtttac
     3421 tttacgagta tttcacagtg tacaatgaac tcacgaaagt taagtatgtc actgagggca
     3481 tgcgtaaacc cgcctttcta agcggagaac agaagaaagc aatagtagat ctgttattca
     3541 agaccaaccg caaagtgaca gttaagcaat tgaaagagga ctactttaag aaaattgaat
     3601 gcttcgattc tgtcgagatc tccggggtag aagatcgatt taatgcgtca cttggtacgt
     3661 atcatgacct cctaaagata attaaagata aggacttcct ggataacgaa gagaatgaag
     3721 atatcttaga agatatagtg ttgactctta ccctctttga agatcgggaa atgattgagg
     3781 aaagactaaa aacatacgct cacctgttcg acgataaggt tatgaaacag ttaaagaggc
     3841 gtcgctatac gggctgggga cgattgtcgc ggaaacttat caacgggata agagacaagc
     3901 aaagtggtaa aactattctc gattttctaa agagcgacgg cttcgccaat aggaacttta
     3961 tgcagctgat ccatgatgac tctttaacct tcaaagagga tatacaaaag gcacaggttt
     4021 ccggacaagg ggactcattg cacgaacata ttgcgaatct tgctggttcg ccagccatca
     4081 aaaagggcat actccagaca gtcaaagtag tggatgagct agttaaggtc atgggacgtc
     4141 acaaaccgga aaacattgta atcgagatgg cacgcgaaaa tcaaacgact cagaaggggc
     4201 aaaaaaacag tcgagagcgg atgaagagaa tagaagaggg tattaaagaa ctgggcagcc
     4261 agatcttaaa ggagcatcct gtggaaaata cccaattgca gaacgagaaa ctttacctct
     4321 attacctaca aaatggaagg gacatgtatg ttgatcagga actggacata aaccgtttat
     4381 ctgattacga cgtcgatgcc attgtacccc aatccttttt gaaggacgat tcaatcgaca
     4441 ataaagtgct tacacgctcg gataagaacc gagggaaaag tgacaatgtt ccaagcgagg
     4501 aagtcgtaaa gaaaatgaag aactattggc ggcagctcct aaatgcgaaa ctgataacgc
     4561 aaagaaagtt cgataactta actaaagctg agaggggtgg cttgtctgaa cttgacaagg
     4621 ccggatttat taaacgtcag ctcgtggaaa cccgccaaat cacaaagcat gttgcacaga
     4681 tactagattc ccgaatgaat acgaaatacg acgagaacga taagctgatt cgggaagtca
     4741 aagtaatcac tttaaagtca aaattggtgt cggacttcag aaaggatttt caattctata
     4801 aagttaggga gataaataac taccaccatg cgcacgacgc ttatcttaat gccgtcgtag
     4861 ggaccgcact cattaagaaa tacccgaagc tagaaagtga gtttgtgtat ggtgattaca
     4921 aagtttatga cgtccgtaag atgatcgcga aaagcgaaca ggagataggc aaggctacag
     4981 ccaaatactt cttttattct aacattatga atttctttaa gacggaaatc actctggcaa
     5041 acggagagat acgcaaacga cctttaattg aaaccaatgg ggagacaggt gaaatcgtat
     5101 gggataaggg ccgggacttc gcgacggtga gaaaagtttt gtccatgccc caagtcaaca
     5161 tagtaaagaa aactgaggtg cagaccggag ggttttcaaa ggaatcgatt cttccaaaaa
     5221 ggaatagtga taagctcatc gctcgtaaaa aggactggga cccgaaaaag tacggtggct
     5281 tcgatagccc tacagttgcc tattctgtcc tagtagtggc aaaagttgag aagggaaaat
     5341 ccaagaaact gaagtcagtc aaagaattat tggggataac gattatggag cgctcgtctt
     5401 ttgaaaagaa ccccatcgac ttccttgagg cgaaaggtta caaggaagta aaaaaggatc
     5461 tcataattaa actaccaaag tatagtctgt ttgagttaga aaatggccga aaacggatgt
     5521 tggctagcgc cggagagctt caaaagggga acgaactcgc actaccgtct aaatacgtga
     5581 atttcctgta tttagcgtcc cattacgaga agttgaaagg ttcacctgaa gataacgaac
     5641 agaagcaact ttttgttgag cagcacaaac attatctcga cgaaatcata gagcaaattt
     5701 cggaattcag taagagagtc atcctagctg atgccaatct ggacaaagta ttaagcgcat
     5761 acaacaagca cagggataaa cccatacgtg agcaggcgga aaatattatc catttgttta
     5821 ctcttaccaa cctcggcgct ccagccgcat tcaagtattt tgacacaacg atagatcgca
     5881 aacgatacac ttctaccaag gaggtgctag acgcgacact gattcaccaa tccatcacgg
     5941 gattatatga aactcggata gatttgtcac agcttggggg tgacggatcc cccaagaaga
     6001 agaggaaagt ctcgagcgac tacaaagacc atgacggtga ttataaagat catgacatcg
     6061 attacaagga tgacgatgac aagtgagaat tcctagagct cgctgatcag cctcgactgt
     6121 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     6181 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     6241 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     6301 agagaatagc aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg agttggccac
     6361 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     6421 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     6481 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     6541 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6601 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6661 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6721 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     6781 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6841 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     6901 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6961 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     7021 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     7081 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     7141 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     7201 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     7261 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     7321 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     7381 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7441 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     7501 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7561 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7621 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     7681 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     7741 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7801 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7861 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7921 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7981 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     8041 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     8101 cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     8161 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     8221 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     8281 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     8341 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     8401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     8461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     8521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     8581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     8641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     8701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     8761 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     8821 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8881 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8941 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     9001 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     9061 gctcacatgt
//
