LOCUS       sequence-364405-       11385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive CRISPR interference (CRISPRi) knock in construct into 
            the AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11385)
  AUTHORS   Mandegar MA, Huebsch N, Frolov EB, Shin E, Truong A, Olvera MP, Chan
            AH, Miyaoka Y, Holmes K, Spencer CI, Judge LM, Gordon DE, Eskildsen 
            TV, Villalta JE, Horlbeck MA, Gilbert LA, Krogan NJ, Sheikh SP, 
            Weissman JS, Qi LS, So PL, Conklin BR
  TITLE     CRISPR Interference Efficiently Induces Specific and Reversible Gene
            Silencing in Human iPSCs.
  JOURNAL   Cell Stem Cell. 2016 Apr 7;18(4):541-53. doi: 
            10.1016/j.stem.2016.01.022. Epub 2016 Mar 10.
  PUBMED    26971820
REFERENCE   2  (bases 1 to 11385)
  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.stem.2016.01.022"; journalName: "Cell Stem Cell"; date: 
            "2016-04-7- 7"; volume: "18"; issue: "4"; pages: "541-53"
COMMENT     Sequence Label: pAAVS1-NC-CRISPRi (Gen3)
FEATURES             Location/Qualifiers
     source          1..11385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          113..1129
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1053..1075
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1137..1156
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1188..1197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1224..1409
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             join(1431..1457,1458..1460,1461..3947,3948..3950,
                     3951..5534)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1431..5534
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5538..5564
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5544..5570
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             5583..5603
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5610..5630
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5761..5780)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5826..5881
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5880..5899)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    complement(6230..6263)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6304..7140
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     rep_origin      complement(7188..7776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7268..7287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7947..8738,8739..8807))
                     /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     8570..8589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_feature    8861..9664
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    complement(9666..9699)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    9706..9731
                     /label=SA
                     /note="splice acceptor site"
     CDS             9755..9808
                     /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"
     regulatory      9824..9833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9830..10633
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9893..9912)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     10503..10522
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(10653..10670)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10659..10883
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        10889..11268
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
       61 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      121 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
      181 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      241 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
      301 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt
      361 gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg
      421 cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc
      481 cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg
      541 ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc
      601 ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg
      661 gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc
      721 tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg
      781 aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact
      841 tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag
      901 cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg
      961 tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga
     1021 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     1081 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     1141 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa tgtcgacgcc accatggatg
     1201 ctaagtcact aactgcctgg tcccggacac tggtgacctt caaggatgta tttgtggact
     1261 tcaccaggga ggagtggaag ctgctggaca ctgctcagca gatcgtgtac agaaatgtga
     1321 tgctggagaa ctataagaac ctggtttcct tgggttatca gcttactaag ccagatgtga
     1381 tcctccggtt ggagaaggga gaagagcccg gcggttccgg cggagggtcg atggacaaga
     1441 agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg
     1501 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     1561 agaagaacct gatcggcgcc ctgctgttcg acagcggaga aacagccgag gccacccggc
     1621 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     1681 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     1741 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     1801 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     1861 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     1921 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     1981 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     2041 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     2101 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc
     2161 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     2221 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     2281 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     2341 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     2401 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     2461 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacatcg
     2521 atggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     2581 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     2641 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     2701 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     2761 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     2821 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     2881 agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     2941 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tacaacgagc
     3001 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     3061 agaaaaaagc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     3121 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     3181 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     3241 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     3301 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     3361 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     3421 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     3481 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     3541 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     3601 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     3661 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     3721 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     3781 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     3841 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     3901 tggaccagga actggacatc aaccggctgt ccgactacga tgtggacgct atcgtgcctc
     3961 agagctttct gaaggacgac tccatcgata acaaagtgct gactcggagc gacaagaacc
     4021 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     4081 gccagctgct gaatgccaag ctgattaccc agaggaagtt cgacaatctg accaaggccg
     4141 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     4201 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     4261 acgagaacga caaactgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     4321 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     4381 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     4441 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     4501 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     4561 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     4621 agacaaacgg cgaaacaggc gagatcgtgt gggataaggg ccgggacttt gccaccgtgc
     4681 ggaaagtgct gtctatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     4741 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga caagctgatc gccagaaaga
     4801 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     4861 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     4921 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     4981 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     5041 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
     5101 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     5161 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaaac
     5221 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     5281 acgctaatct ggacaaggtg ctgagcgcct acaacaagca cagagacaag cctatcagag
     5341 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     5401 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
     5461 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     5521 agctgggagg cgacgcctat ccctatgacg tgcccgatta tgccagcctg ggcagcggct
     5581 cccccaagaa aaaacgcaag gtggaagatc ctaagaaaaa gcggaaagtg gacggcattg
     5641 gtagtgggag caacggcagc agcggatcct aacgcgtgaa ttcactcctc aggtgcaggc
     5701 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     5761 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     5821 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     5881 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     5941 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     6001 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     6061 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     6121 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     6181 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagctta taacttcgta
     6241 tagcatacat tatacgaagt tatgcggccg caccggtatg catttaatta agatatcagc
     6301 tttactaggg acaggattgg tgacagaaaa gccccatcct taggcctcct ccttcctagt
     6361 ctcctgatat tgggtctaac ccccacctcc tgttaggcag attccttatc tggtgacaca
     6421 cccccatttc ctggagccat ctctctcctt gccagaacct ctaaggtttg cttacgatgg
     6481 agccagagag gatcctggga gggagagctt ggcagggggt gggagggaag ggggggatgc
     6541 gtgacctgcc cggttctcag tggccaccct gcgctaccct ctcccagaac ctgagctgct
     6601 ctgacgcggc tgtctggtgc gtttcactga tcctggtgct gcagcttcct tacacttccc
     6661 aagaggagaa gcagtttgga aaaacaaaat cagaataagt tggtcctgag ttctaacttt
     6721 ggctcttcac ctttctagtc cccaatttat attgttcctc cgtgcgtcag ttttacctgt
     6781 gagataaggc cagtagccag ccccgtcctg gcagggctgt ggtgaggagg ggggtgtccg
     6841 tgtggaaaac tccctttgtg agaatggtgc gtcctaggtg ttcaccaggt cgtggccgcc
     6901 tctactccct ttctctttct ccatccttct ttccttaaag agtccccagt gctatctggg
     6961 acatattcct ccgcccagag cagggtcccg cttccctaag gccctgctct gggcttctgg
     7021 gtttgagtcc ttggcaagcc caggagaggc gctcaggctt ccctgtcccc cttcctcgtc
     7081 caccatctca tgcccctggc tctcctgccc cttccctaca ggggttcctg gctctgctct
     7141 aagggcgaat tcgccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7201 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7261 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7321 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7381 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7441 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7501 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7561 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7621 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7681 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7741 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7801 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7861 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7921 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7981 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     8041 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     8101 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     8161 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8221 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8281 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8341 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8401 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8461 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8521 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8581 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8641 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8701 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8761 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8821 caatattatt gaagcattta tcagggttga attcgccctt tgctttctct gaccagcatt
     8881 ctctcccctg ggcctgtgcc gctttctgtc tgcagcttgt ggcctgggtc acctctacgg
     8941 ctggcccaga tccttccctg ccgcctcctt caggttccgt cttcctccac tccctcttcc
     9001 ccttgctctc tgctgtgttg ctgcccaagg atgctctttc cggagcactt ccttctcggc
     9061 gctgcaccac gtgatgtcct ctgagcggat cctccccgtg tctgggtcct ctccgggcat
     9121 ctctcctccc tcacccaacc ccatgccgtc ttcactcgct gggttccctt ttccttctcc
     9181 ttctggggcc tgtgccatct ctcgtttctt aggatggcct tctccgacgg atgtctccct
     9241 tgcgtcccgc ctccccttct tgtaggcctg catcatcacc gtttttctgg acaaccccaa
     9301 agtaccccgt ctccctggct ttagccacct ctccatcctc ttgctttctt tgcctggaca
     9361 ccccgttctc ctgtggattc gggtcacctc tcactccttt catttgggca gctcccctac
     9421 cccccttacc tctctagtct gtgctagctc ttccagcccc ctgtcatggc atcttccagg
     9481 ggtccgagag ctcagctagt cttcttcctc caacccgggc ccctatgtcc acttcaggac
     9541 agcatgtttg ctgcctccag ggatcctgtg tccccgagct gggaccacct tatattccca
     9601 gggccggtta atgtggctct ggttctgggt acttttatct gtcccctcca ccccacagtg
     9661 gggcaataac ttcgtatagc atacattata cgaagttata agcttctgac ctcttctctt
     9721 cctcccacag ggcctcgaga gatctggcag cggagagggc agaggaagtc ttctaacatg
     9781 cggtgacgtg gaggagaatc ccggccctag gctcgagggt accgccacca tgggatcggc
     9841 cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     9901 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     9961 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
    10021 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
    10081 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
    10141 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
    10201 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
    10261 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
    10321 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
    10381 cgatgatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
    10441 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
    10501 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
    10561 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
    10621 cgagttcttc tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
    10681 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
    10741 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
    10801 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
    10861 tggggatgcg gtgggctcta tgggtctcga cattgattat tgactagtta ttaatagtaa
    10921 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
    10981 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
    11041 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
    11101 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
    11161 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
    11221 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc
    11281 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
    11341 tttatttttt aattattttg tgcagcgatg ggggcggggg ggggg
//
