LOCUS       sequence-361180-        9277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pU6-MS2-gRNA-backbone(BbsI)-CBH-dSpCas9-T2A-EBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9277)
  AUTHORS   Wang C, Qu Y, Cheng JKW, Hughes NW, Zhang Q, Wang M, Cong L
  TITLE     dCas9-based gene editing for cleavage-free genomic knock-in of long 
            sequences.
  JOURNAL   Nat Cell Biol. 2022 Feb;24(2):268-278. doi: 
            10.1038/s41556-021-00836-1. Epub 2022 Feb 10.
  PUBMED    35145221
REFERENCE   2  (bases 1 to 9277)
  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/s41556-021-00836-1"; journalName: "Nat Cell Biol"; date: 
            "2022-02"; volume: "24"; issue: "2"; pages: "268-278"
COMMENT     Sequence Label: p-dCas9-SSAP-MS2-BB_BbsI
FEATURES             Location/Qualifiers
     source          1..9277
                     /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        285..303
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        378..396
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     enhancer        417..702
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        704..981
                     /label=chicken beta-actin promoter
     intron          982..1209
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1221..1230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1230..1295
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1302..1322
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1347..1370,1371..1373,1374..3929,3930..3932,
                     3933..5447)
                     /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 N863A 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             join(1347..1370,1371..1373,1374..3860,3861..3863,
                     3864..5447)
                     /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             5448..5495
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5511..5564
                     /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             join(5571..5573,5574..5576,5577..6290)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(6318..6335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6324..6531
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6540..6680
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6540..6669
                     /label=AAV2 ITR
     rep_origin      6755..7210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6842..6861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7052..7073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7227..7246)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7346..7368
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7406..7424)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7492..7596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7597..7665,7666..8457)
                     /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(7815..7834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8628..9216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9117..9136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtggcgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt taagagctag gccaacatga ggatcaccca
      301 tgtctgcagg gcctagcaag tttaaataag gctagtccgt tatcaacttg aaaaagtggc
      361 accgagtcgg tgcgcgcaca tgaggatcac ccatgtgctt tttttctaga ggtacccgtt
      421 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      481 tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      541 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      601 aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct
      661 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
      721 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
      781 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccrggsggg
      841 gsggggsggg gsgrggggsg gggsggggcg aggcggagag gtgcggcggc agccaatcag
      901 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
      961 aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc
     1021 gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc
     1081 gggcgggacg gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag
     1141 ggatggttgg ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact
     1201 ttttttcagg ttggaccggt gccaccatgg actataagga ccacgacgga gactacaagg
     1261 atcatgatat tgattacaaa gacgatgacg ataagatggc cccaaagaag aagcggaagg
     1321 tcggtatcca cggagtccca gcagccgaca agaagtacag catcggcctg gccatcggca
     1381 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
     1441 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt
     1501 tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
     1561 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
     1621 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
     1681 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     1741 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
     1801 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     1861 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     1921 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     1981 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     2041 agaagaatgg cctgttcggc aacctgattg ccctgagcct gggcctgacc cccaacttca
     2101 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
     2161 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     2221 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     2281 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
     2341 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     2401 accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct
     2461 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     2521 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
     2581 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     2641 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     2701 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     2761 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg
     2821 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     2881 tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg
     2941 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt
     3001 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
     3061 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
     3121 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
     3181 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
     3241 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
     3301 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
     3361 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
     3421 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
     3481 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
     3541 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
     3601 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
     3661 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
     3721 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
     3781 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
     3841 tgtccgacta cgatgtggac gccatcgtgc ctcagagctt tctgaaggac gactccatcg
     3901 acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg
     3961 aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta
     4021 cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
     4081 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
     4141 agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag
     4201 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
     4261 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
     4321 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
     4381 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
     4441 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
     4501 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg
     4561 tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga
     4621 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
     4681 agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
     4741 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
     4801 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
     4861 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
     4921 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
     4981 tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
     5041 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
     5101 agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga
     5161 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg
     5221 cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt
     5281 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
     5341 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
     5401 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg
     5461 ccacgaaaaa ggccggccag gcaaaaaaga aaaaggaatt cggcagtgga gagggcagag
     5521 gaagtctgct aacatgcggt gacgtggagg agaatcccgg ccctgctagc atggtgagca
     5581 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     5641 acggccacaa gttcagcgtg aggggcgagg gcgagggcga tgccaccaac ggcaagctga
     5701 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     5761 ccctgagcca cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     5821 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     5881 acggcaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     5941 tcgagctgaa gggcgtcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     6001 acaacttcaa cagccacaac atctatatca tggccgtcaa gcagaagaac ggcatcaagg
     6061 tgaacttcaa gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc
     6121 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacagccac tacctgagca
     6181 cccagtccgt gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     6241 tccgcaccgc cgccgggatc actctcggca tggacgagct gtacaagtga gaattctaac
     6301 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     6361 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     6421 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     6481 gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg agcggccgca
     6541 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     6601 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     6661 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     6721 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     6781 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     6841 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     6901 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     6961 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     7021 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     7081 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     7141 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     7201 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     7261 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     7321 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     7381 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     7441 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     7501 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7561 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7621 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7681 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7741 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7801 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7861 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7921 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7981 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     8041 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     8101 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     8161 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     8221 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     8281 attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg
     8341 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     8401 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8461 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8521 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8581 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8641 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8701 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8761 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8821 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8881 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8941 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     9001 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     9061 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     9121 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     9181 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     9241 cggttcctgg ccttttgctg gccttttgct cacatgt
//
