LOCUS       sequence-170272-       10347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. pyogenes FokI-dCas9-NLS with a 25 amino acid linker 
            (GGGGS)5 fusion. Also expresses Csy4 for cleavage of multiplexed 
            gRNA transcripts. Derived from pSQT1601 (Addgene #53369)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10347)
  AUTHORS   Havlicek S, Shen Y, Alpagu Y, Bruntraeger MB, Zufir NB, Phuah ZY, Fu
            Z, Dunn NR, Stanton LW
  TITLE     Re-engineered RNA-Guided FokI-Nucleases for Improved Genome Editing 
            in Human Cells.
  JOURNAL   Mol Ther. 2017 Feb 1;25(2):342-355. doi: 
            10.1016/j.ymthe.2016.11.007.
  PUBMED    28153087
REFERENCE   2  (bases 1 to 10347)
  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: 
            "10.1016/j.ymthe.2016.11"; journalName: "Mol Ther"; date: 
            "2017-02-1- 1"; volume: "25"; issue: "2"; pages: "342-355"
COMMENT     Sequence Label: pSH-Csy4-T2A-SpRFN
FEATURES             Location/Qualifiers
     source          1..10347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          160..1168
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1092..1114
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1176..1195
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1261..1270
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1831..1884
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1888..1908
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1915..2502
                     /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(2578..5976,5977..5979,5980..6576,6577,6578..6580,
                     6581..6582,6583..6585,6586..6678)
                     /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(2578..2601,2602..2604,2605..5091,5092..5094,
                     5095..6678)
                     /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             6685..6705
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6712..6777
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6867..6886)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6932..6987
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6986..7005)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7348..7364)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7348..7364)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7361..7383)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7372..7388
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7396..7402,7403..7420,7421..7426))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7441..7462
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7521..7716
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7567..7702
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7722..7856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7759..7778)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7813..7832
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7924..7941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8095..8683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8175..8194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8854..9645,9646..9714))
                     /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     9477..9496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9715..9819)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        9849..10228
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(10231..10347,1..159)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
       61 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      121 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gttgccttcg
      181 ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      241 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      301 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc
      361 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
      421 cgcgtgcggc ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
      481 gcgctccgcg tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
      541 ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt
      601 gggcgcggcg gtcgggctgt aacccccccc tgcacccccc tccccgagtt gctgagcacg
      661 gcccggcttc gggtgcgggg ctccgtgcgg ggcgtggcgc ggggctcgcc gtgccgggcg
      721 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
      781 cgggggaggg gcgcggcggc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag
      841 ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg
      901 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg gcgaagcggt
      961 gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
     1021 cccttctcca tctccagcct cggggctgcc gcagggggac ggctgccttc gggggggacg
     1081 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
     1141 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
     1201 tcatcatttt ggcaaagaat tctgcagtcg acggtaccgc gggcccggga tccaccggtc
     1261 gccaccatgg gtgatcatta tctggatatt cggctgaggc ctgatccaga gttcccacct
     1321 gcgcagctga tgtctgtcct ttttggcaaa cttcatcagg ccctggttgc ccagggcgga
     1381 gatcggatag gggtaagctt tccagacctc gacgaaagcc ggagccgcct gggagaacgc
     1441 ctgcggatcc acgcttctgc cgacgatctg agagccttgc tggcaaggcc atggcttgag
     1501 gggctccggg atcacctgca gtttggcgaa cccgccgttg ttccccaccc aaccccttat
     1561 cggcaggtgt ctagagtgca ggccaaatct aatccagaac ggctgcgacg gcgactcatg
     1621 cggcgacatg atcttagcga ggaagaggcc cgaaaaagaa tccctgatac cgtggcccgc
     1681 gcccttgact tgccttttgt cacactgcgg tcccagagta cggggcagca tttcagactt
     1741 ttcattcgac acgggccact gcaagttacc gccgaagaag gaggctttac ttgttatgga
     1801 ctctccaagg gaggtttcgt gccctggttt gagggcagag gaagtctgtt aacatgcggt
     1861 gacgtcgagg agaatcctgg cccaatgcct aagaagaagc ggaaggtgag cagccaactt
     1921 gtgaagtctg aactcgagga gaaaaaatca gagttgagac acaagttgaa gtacgtgcca
     1981 cacgaataca tcgagcttat cgagatcgcc agaaacagta cccaggatag gatccttgag
     2041 atgaaagtca tggagttctt tatgaaggtc tacggttata gaggaaagca ccttggcggt
     2101 agcagaaagc ccgatggcgc catctatact gtcggatctc ctatcgatta tggggtgatc
     2161 gtggatacca aagcttactc aggcgggtac aacttgccca taggacaagc cgacgagatg
     2221 cagcggtatg tcgaagagaa ccagacgcgc aacaagcaca tcaaccccaa tgaatggtgg
     2281 aaagtgtacc caagtagtgt gactgagttc aagttcctgt ttgtctccgg ccactttaag
     2341 ggcaattata aagctcagct cactagactc aatcacatca caaactgcaa cggagctgtg
     2401 ttgtcagtgg aggagctcct gattggaggc gagatgatca aagccggcac ccttacactg
     2461 gaggaggtgc ggcggaagtt caacaatgga gagatcaact tcggtggcgg tggatccggt
     2521 ggcggtggat ccggtggcgg tggatccggt ggcggtggat ccggtggtgg tggatccgat
     2581 aaaaagtatt ctattggttt agccatcggc actaattccg ttggatgggc tgtcataacc
     2641 gatgaataca aagtaccttc aaagaaattt aaggtgttgg ggaacacaga ccgtcattcg
     2701 attaaaaaga atcttatcgg tgccctccta ttcgatagtg gcgaaacggc agaggcgact
     2761 cgcctgaaac gaaccgctcg gagaaggtat acacgtcgca agaaccgaat atgttactta
     2821 caagaaattt ttagcaatga gatggccaaa gttgacgatt ctttctttca ccgtttggaa
     2881 gagtccttcc ttgtcgaaga ggacaagaaa catgaacggc accccatctt tggaaacata
     2941 gtagatgagg tggcatatca tgaaaagtac ccaacgattt atcacctcag aaaaaagcta
     3001 gttgactcaa ctgataaagc ggacctgagg ttaatctact tggctcttgc ccatatgata
     3061 aagttccgtg ggcactttct cattgagggt gatctaaatc cggacaactc ggatgtcgac
     3121 aaactgttca tccagttagt acaaacctat aatcagttgt ttgaagagaa ccctataaat
     3181 gcaagtggcg tggatgcgaa ggctattctt agcgcccgcc tctctaaatc ccgacggcta
     3241 gaaaacctga tcgcacaatt acccggagag aagaaaaatg ggttgttcgg taaccttata
     3301 gcgctctcac taggcctgac accaaatttt aagtcgaact tcgacttagc tgaagatgcc
     3361 aaattgcagc ttagtaagga cacgtacgat gacgatctcg acaatctact ggcacaaatt
     3421 ggagatcagt atgcggactt atttttggct gccaaaaacc ttagcgatgc aatcctccta
     3481 tctgacatac tgagagttaa tactgagatt accaaggcgc cgttatccgc ttcaatgatc
     3541 aaaaggtacg atgaacatca ccaagacttg acacttctca aggccctagt ccgtcagcaa
     3601 ctgcctgaga aatataagga aatattcttt gatcagtcga aaaacgggta cgcaggttat
     3661 attgacggcg gagcgagtca agaggaattc tacaagttta tcaaacccat attagagaag
     3721 atggatggga cggaagagtt gcttgtaaaa ctcaatcgcg aagatctact gcgaaagcag
     3781 cggactttcg acaacggtag cattccacat caaatccact taggcgaatt gcatgctata
     3841 cttagaaggc aggaggattt ttatccgttc ctcaaagaca atcgtgaaaa gattgagaaa
     3901 atcctaacct ttcgcatacc ttactatgtg ggacccctgg cccgagggaa ctctcggttc
     3961 gcatggatga caagaaagtc cgaagaaacg attactccat ggaattttga ggaagttgtc
     4021 gataaaggtg cgtcagctca atcgttcatc gagaggatga ccaactttga caagaattta
     4081 ccgaacgaaa aagtattgcc taagcacagt ttactttacg agtatttcac agtgtacaat
     4141 gaactcacga aagttaagta tgtcactgag ggcatgcgta aacccgcctt tctaagcgga
     4201 gaacagaaga aagcaatagt agatctgtta ttcaagacca accgcaaagt gacagttaag
     4261 caattgaaag aggactactt taagaaaatt gaatgcttcg attctgtcga gatctccggg
     4321 gtagaagatc gatttaatgc gtcacttggt acgtatcatg acctcctaaa gataattaaa
     4381 gataaggact tcctggataa cgaagagaat gaagatatct tagaagatat agtgttgact
     4441 cttaccctct ttgaagatcg ggaaatgatt gaggaaagac taaaaacata cgctcacctg
     4501 ttcgacgata aggttatgaa acagttaaag aggcgtcgct atacgggctg gggacgattg
     4561 tcgcggaaac ttatcaacgg gataagagac aagcaaagtg gtaaaactat tctcgatttt
     4621 ctaaagagcg acggcttcgc caataggaac tttatgcagc tgatccatga tgactcttta
     4681 accttcaaag aggatataca aaaggcacag gtttccggac aaggggactc attgcacgaa
     4741 catattgcga atcttgctgg ttcgccagcc atcaaaaagg gcatactcca gacagtcaaa
     4801 gtagtggatg agctagttaa ggtcatggga cgtcacaaac cggaaaacat tgtaatcgag
     4861 atggcacgcg aaaatcaaac gactcagaag gggcaaaaaa acagtcgaga gcggatgaag
     4921 agaatagaag agggtattaa agaactgggc agccagatct taaaggagca tcctgtggaa
     4981 aatacccaat tgcagaacga gaaactttac ctctattacc tacaaaatgg aagggacatg
     5041 tatgttgatc aggaactgga cataaaccgt ttatctgatt acgacgtcga tgccattgta
     5101 ccccaatcct ttttgaagga cgattcaatc gacaataaag tgcttacacg ctcggataag
     5161 aaccgaggga aaagtgacaa tgttccaagc gaggaagtcg taaagaaaat gaagaactat
     5221 tggcggcagc tcctaaatgc gaaactgata acgcaaagaa agttcgataa cttaactaaa
     5281 gctgagaggg gtggcttgtc tgaacttgac aaggccggat ttattaaacg tcagctcgtg
     5341 gaaacccgcc aaatcacaaa gcatgttgca cagatactag attcccgaat gaatacgaaa
     5401 tacgacgaga acgataagct gattcgggaa gtcaaagtaa tcactttaaa gtcaaaattg
     5461 gtgtcggact tcagaaagga ttttcaattc tataaagtta gggagataaa taactaccac
     5521 catgcgcacg acgcttatct taatgccgtc gtagggaccg cactcattaa gaaatacccg
     5581 aagctagaaa gtgagtttgt gtatggtgat tacaaagttt atgacgtccg taagatgatc
     5641 gcgaaaagcg aacaggagat aggcaaggct acagccaaat acttctttta ttctaacatt
     5701 atgaatttct ttaagacgga aatcactctg gcaaacggag agatacgcaa acgaccttta
     5761 attgaaacca atggggagac aggtgaaatc gtatgggata agggccggga cttcgcgacg
     5821 gtgagaaaag ttttgtccat gccccaagtc aacatagtaa agaaaactga ggtgcagacc
     5881 ggagggtttt caaaggaatc gattcttcca aaaaggaata gtgataagct catcgctcgt
     5941 aaaaaggact gggacccgaa aaagtacggt ggcttcgata gccctacagt tgcctattct
     6001 gtcctagtag tggcaaaagt tgagaaggga aaatccaaga aactgaagtc agtcaaagaa
     6061 ttattgggga taacgattat ggagcgctcg tcttttgaaa agaaccccat cgacttcctt
     6121 gaggcgaaag gttacaagga agtaaaaaag gatctcataa ttaaactacc aaagtatagt
     6181 ctgtttgagt tagaaaatgg ccgaaaacgg atgttggcta gcgccggaga gcttcaaaag
     6241 gggaacgaac tcgcactacc gtctaaatac gtgaatttcc tgtatttagc gtcccattac
     6301 gagaagttga aaggttcacc tgaagataac gaacagaagc aactttttgt tgagcagcac
     6361 aaacattatc tcgacgaaat catagagcaa atttcggaat tcagtaagag agtcatccta
     6421 gctgatgcca atctggacaa agtattaagc gcatacaaca agcacaggga taaacccata
     6481 cgtgagcagg cggaaaatat tatccatttg tttactctta ccaacctcgg cgctccagcc
     6541 gcattcaagt attttgacac aacgatagat cgcaaacgat acacttctac caaggaggtg
     6601 ctagacgcga cactgattca ccaatccatc acgggattat atgaaactcg gatagatttg
     6661 tcacagcttg ggggtgacgg atcccccaag aagaagagga aagtctcgag cgactacaaa
     6721 gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga tgacaagtga
     6781 agcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     6841 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     6901 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     6961 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     7021 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     7081 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     7141 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     7201 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     7261 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     7321 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7381 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7441 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7501 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     7561 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     7621 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     7681 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     7741 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     7801 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     7861 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7921 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7981 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     8041 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     8101 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8161 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8221 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8281 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8341 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8401 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8461 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8521 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8581 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8641 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8701 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8761 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8821 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8881 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8941 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     9001 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     9061 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     9121 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9181 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     9241 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9301 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9361 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9421 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9481 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9541 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9601 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9661 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9721 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9781 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9841 acctggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     9901 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     9961 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
    10021 aatagggact ttccattgac gtcaatgggt ggactattta cggtaaactg cccacttggc
    10081 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
    10141 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
    10201 ctacgtatta gtcatcgcta ttaccatggg tcgaggtgag ccccacgttc tgcttcactc
    10261 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
    10321 gtgcagcgat gggggcgggg ggggggg
//
