LOCUS       sequence-434913-       14075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible (Zim3)KRAB-dCas9-HA-P2A-mCherry cassette for 
            integration at the AAVS1-locus of the gorilla genome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14075)
  AUTHORS   Edenhofer FC, Termeg A, Ohnuki M, Jocher J, Kliesmete Z, Briem E, 
            Hellmann I, Enard W
  TITLE     Generation and characterization of inducible KRAB-dCas9 iPSCs from 
            primates for cross-species CRISPRi.
  JOURNAL   iScience. 2024 May 23;27(6):110090. doi: 10.1016/j.isci.2024.110090.
            eCollection 2024 Jun 21.
  PUBMED    38947524
REFERENCE   2  (bases 1 to 14075)
  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: "iScience.";
            date: "2024-05-23"; volume: "27(6):110090. doi"; pages: " 
            10.1016/j.isci.2024.110090. eCollection 2024 Jun 21"
COMMENT     Sequence Label: pGorilla-AAVS1-TetOn-ZIM3-KRAB-dCas9-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..14075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    14..32
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    50..68
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    86..104
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    122..140
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    158..176
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    194..212
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    230..248
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     293..317
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    399..423
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(772..796)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             816..836
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      885..894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(891..917,918..920,921..3407,3408..3410,3411..4994)
                     /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             891..4994
                     /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             4998..5024
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5004..5030
                     /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             5043..5063
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5070..5090
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5109..5165
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5166..5876
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5311..5329)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5579..5598
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5908..6029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5945..5964)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5999..6018
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6078..6094)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6078..6094)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6091..6113)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6102..6118
                     /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(6126..6132,6133..6150,6151..6156))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6171..6192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6309..6326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6382..6415
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     6746..6765
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    6764..6819
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6865..6884
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(6971..7717)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(7749..7768)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          7776..8792
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(7830..7852)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        9072..9451
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    9457..9681
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     9670..9687
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(9707..10510)
                     /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(9818..9837)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     10428..10447
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     regulatory      10507..10516
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(10532..10585)
                     /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"
     misc_feature    10609..10634
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    10684..11491
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     CDS             join(11545..11613,11614..12405)
                     /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(11763..11782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12576..13164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13065..13084
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    13212..14048
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
ORIGIN
        1 cgcgccggtt tactccctat cagtgataga gaacgtatga agagtttact ccctatcagt
       61 gatagagaac gtatgcagac tttactccct atcagtgata gagaacgtat aaggagttta
      121 ctccctatca gtgatagaga acgtatgacc agtttactcc ctatcagtga tagagaacgt
      181 atctacagtt tactccctat cagtgataga gaacgtatat ccagtttact ccctatcagt
      241 gatagagaac gtataagctt taggcgtgta cggtgggcgc ctataaaagc agagctcgtt
      301 tagtgaaccg tcagatcgcc tggagcaatt ccacaacact tttgtcttat accaactttc
      361 cgtaccactt cctaccctcg taaacttaag gttaatcaac aagtttgtac aaaaaagcag
      421 gctggcgccg gaaccaattc agtcgacagg cctaggctac gcgcgccacc atgaacaatt
      481 cccagggaag agtgaccttc gaggatgtca ctgtgaactt cacccagggg gagtggcagc
      541 ggctgaatcc cgaacagaga aacttgtaca gggatgtgat gctggagaat tacagcaacc
      601 ttgtctctgt gggacaaggg gaaaccacca aacccgatgt gatcttgagg ttggaacaag
      661 gaaaggagcc atggttggag gaagaggaag tgctgggaag tggccgtgca gaaaaaaatg
      721 gggacattgg agggcagatt tggaagccaa aggatgtgaa agagagtctc tacccaactt
      781 tcttgtacaa agtggttggt ggatctggag gttctcccaa gaagaagcgc aaggtgggac
      841 gcgtctgcag gatatcaagc ttgcggtacc gcgggcccgg gatcgccacc atggacaaga
      901 agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg
      961 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     1021 agaagaacct gatcggcgcc ctgctgttcg acagcggaga aacagccgag gccacccggc
     1081 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     1141 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     1201 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     1261 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     1321 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     1381 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     1441 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     1501 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     1561 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc
     1621 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     1681 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     1741 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     1801 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     1861 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     1921 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacatcg
     1981 atggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     2041 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     2101 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     2161 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     2221 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     2281 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     2341 agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     2401 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tacaacgagc
     2461 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     2521 agaaaaaagc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     2581 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     2641 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     2701 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     2761 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     2821 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     2881 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     2941 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     3001 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     3061 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     3121 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     3181 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     3241 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     3301 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     3361 tggaccagga actggacatc aaccggctgt ccgactacga tgtggacgct atcgtgcctc
     3421 agagctttct gaaggacgac tccatcgata acaaagtgct gactcggagc gacaagaacc
     3481 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     3541 gccagctgct gaatgccaag ctgattaccc agaggaagtt cgacaatctg accaaggccg
     3601 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     3661 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     3721 acgagaacga caaactgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     3781 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     3841 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     3901 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     3961 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     4021 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     4081 agacaaacgg cgaaacaggc gagatcgtgt gggataaggg ccgggacttt gccaccgtgc
     4141 ggaaagtgct gtctatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     4201 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga caagctgatc gccagaaaga
     4261 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     4321 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     4381 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     4441 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     4501 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
     4561 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     4621 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaaac
     4681 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     4741 acgctaatct ggacaaggtg ctgagcgcct acaacaagca cagagacaag cctatcagag
     4801 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     4861 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
     4921 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     4981 agctgggagg cgacgcctat ccctatgacg tgcccgatta tgccagcctg ggcagcggct
     5041 cccccaagaa aaaacgcaag gtggaagatc ctaagaaaaa gcggaaagtg gacggcattg
     5101 gtagtggggc tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg
     5161 gacctatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     5221 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     5281 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     5341 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     5401 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     5461 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     5521 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     5581 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     5641 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     5701 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     5761 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     5821 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaaaccg
     5881 gtgcggccgc aatcgatcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5941 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6001 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6061 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6121 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6181 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6241 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6301 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6361 gctcactcaa aggacggccg cataacttcg tataatgtat gctatacgaa gttataagct
     6421 tgggctgcag gtcgagggat ctccataaga gaagagggac agctatgact gggagtagtc
     6481 aggagaggag gaaaaatctg gctagtaaaa catgtaagga aaattttagg gatgttaaag
     6541 aaaaaaataa cacaaaacaa aatataaaaa aaatctaacc tcaagtcaag gcttttctat
     6601 ggaataagga atggacagca gggggctgtt tcatatactg atgacctctt tatagccaac
     6661 ctttgttcat ggcagccagc atatgggcat atgttgccaa actctaaacc aaatactcat
     6721 tctgatgttt taaatgattt gccctcccat atgtccttcc gagtgagaga cacaaaaaat
     6781 tccaacacac tattgcaatg aaaataaatt tcctttatta gccagaagtc agatgctcaa
     6841 ggggcttcat gatgtcccca taatttttgg cagagggaaa aagatctcag tggtatttgt
     6901 gagccagggc attggccaca ccagccacca ccttctgata ggcagcctgc acctgaggag
     6961 tgaattcatt ttagcctggc agcatatcga gatcgaagtc atcgagagcg tcagcaggga
     7021 gcatatccag atcaaaatca tccagggcat cggcgggcag catgtccagg tcgaagtcgt
     7081 ccagggcgtc ggtgggtccg ccgctctcgc acttcagctg cttctcgagg ccgcagatga
     7141 tcagttccag gccgaacagg aaggcgggct cggcgccctg tctgtcgaac agctcgatgg
     7201 cctgcttcag cagggggggc atgctgtcgg tggtgggtgt ctctctttcc tctttggcga
     7261 cctggtgttc ctgttcttcc agcacgcagc ccagggtgaa gtggcccacg gcgctcaggg
     7321 cgtacagggc gttttccagg ctgaagccct gctggcacag gaaggccagc tggttttcca
     7381 gtgtctcgta ctgcttctcg gtgggtctgg tgcccaggtg cactttggcg ccgtcccggt
     7441 ggctcagcag ggcgcatctg tagctcttgg cgttgttccg caggaagtcc tgccagctct
     7501 cgccttccag agggcagctg tgggtgtggt gccggtccag catctcgatg ggcagggcgt
     7561 ccagcagggc ccgcttgttc ttcacgtgcc agtacagggt gggctgttcc acgcccagtt
     7621 tctgggccag cttccgggtg gtcaggccct cgatgcccac gccgttcagc agttccaggg
     7681 cgctgttgat cactttgctc ttgtccagcc ggctcatgtc gacattcttt gccaaaatga
     7741 tgagacagca caataaccag cacgttgccc aggagctgta ggaaaaagaa gaaggcatga
     7801 acatggttag cagaggctct agagccgccg gtcacacgcc agaagccgaa ccccgccctg
     7861 ccccgtcccc cccgaaggca gccgtccccc cgcggacagc cccgaggctg gagagggaga
     7921 aggggacggc ggcgcggcga cgcacgaagg ccctccccgc ccatttcctt cctgccggcg
     7981 ccgcaccgct tcgccccgcg cccgctagag ggggtgcggc ggcgcctccc agatttcggc
     8041 tccgcacaga tttgggacaa aggaagtccc tgcgccctct cgcacgatta ccataaaagg
     8101 caatggctgc ggctcgccgc gcctcgacag ccgccggcgc tccgggggcc gccgcgcccc
     8161 ycccccgagc ccyccccggc ccgaggcggc cccgccccgc ccggcacccc cacctgccgc
     8221 caccccccgc ccggcacggc gagccccgcg ccacgccccg tacggagccc cgcacccgaa
     8281 gccgggccgt gctcagcaac tcggggaggg gggtgcaggg ggggttgcag cccgaccgac
     8341 gcgcccacac cccctgctca cccccccacg cacacacccc gcacgcagcc tttgttcccc
     8401 tcgcagcccc ccccgcaccg cggggcaccg cccccggccg cgctcccctc gcgcacactg
     8461 cggagcgcac aaagccccgc gccgcgcccg cagcgctcac agccgccggg cagcgcggag
     8521 ccgcacgcgg cgctccccac gcacacacac acgcacgcac cccccgagcc gctccccccg
     8581 cacaaagggc cctcccggag cccctcaagg ctttcacgca gccacagaaa agaaacaagc
     8641 cgtcattaaa ccaagcgcta attacagccc ggaggagaag ggccgtcccg cccgctcacc
     8701 tgtgggagta acgcggtcag tcagagccgg ggcgggcggc gcgaggcggc ggcggagcgg
     8761 ggcacggggc gaaggcagcg cgcagcgact cccgcccgcc gcgcgcttcg ctttttatag
     8821 ggccgccgcc gccgccgcct cgccataaaa ggaaactttc ggagcgcgcc gctctgattg
     8881 gctgccgccg cmccycyccs ccycsccccs ccccsccccc cscccccccc ccccccsccy
     8941 ggcgcgcgcc cccccccccc ccccgccccc atcgctgcac aaaataatta aaaaataaat
     9001 aaatacaaaa ttgggggtgg ggaggggggg gagatgggga gagtgaagca gaacgtgggg
     9061 ctcacctcga ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     9121 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     9181 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     9241 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     9301 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     9361 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
     9421 tactattaat aactagtcaa taatcaatgt cgagacccat agagcccacc gcatccccag
     9481 catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
     9541 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
     9601 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
     9661 gctggcaact agaaggcaca gtcgaggctg atcagcgggt ttaaactcag aagaactcgt
     9721 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     9781 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     9841 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     9901 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcatcgc
     9961 cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
    10021 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
    10081 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
    10141 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
    10201 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
    10261 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct
    10321 gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
    10381 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
    10441 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatgg
    10501 ccgatcccat ggtggcggta ccctcgagcc tagggccggg attctcctcc acgtcaccgc
    10561 atgttagaag acttcctctg ccctctccgc tgccagatct ctcgaggccc tgtgggagga
    10621 agagaagagg tcagaagctt ataacttcgt ggcgcgccat aatgtatgct atacgaagtt
    10681 attgccccac tgtggggtgg aggggacaga taaaagtacc cagaaccaga gccacattaa
    10741 ccggccctgg gaatataagg tggtcccagc tcggagacac aggatccctg gaggaagcaa
    10801 acatgctgtc ctgaagtgga tataggggcc cgggttggag gaagaagact agctgagctc
    10861 tgggacccct ggaacatgcc atgacagggg gctggaagag cgagcacaga ctagagaggt
    10921 aaggggggta ggggagctgc ccaaataaaa ggagtgagag gtgacccaaa tccacaggag
    10981 aacggggtgt ccaggcaaag aaagcaagag gatggagagg tggctaaagc cagggagacg
    11041 gggtactttg gggttgtcca gaaaaaacgg tgatgatgca ggcctacaag aaggggaggc
    11101 ggggacgcaa aggagagaca tccgtcggag aaggccatcc taagaaacga gagatggcac
    11161 aggccccaga aggagaagga aaagggaacc cagcgagtga agacggcatg gggttgggtt
    11221 agagaggaga gatgcccgga gaggacccag acaaggggag gatccgctca gaggacatca
    11281 cgtggtgcag cgccgagaag gaagtgctcc ggaaagagca tccttgggca gcaacacagc
    11341 agagagcaag gggaagaggg agtggaggaa gatggaacct gaaggaggtg gcagggaagg
    11401 atctgggcca gccgtagagg tgacccaggc cacaagctgc agacagaaag cggcacaggc
    11461 ccaggggaga gaatgcaggt cagagaaagc aaagggcgaa ttcaaccctg ataaatgctt
    11521 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
    11581 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
    11641 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
    11701 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
    11761 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
    11821 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
    11881 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
    11941 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
    12001 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
    12061 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
    12121 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
    12181 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
    12241 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
    12301 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
    12361 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
    12421 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
    12481 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
    12541 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
    12601 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
    12661 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
    12721 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
    12781 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
    12841 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
    12901 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
    12961 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
    13021 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
    13081 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
    13141 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcg
    13201 aattcgccct tagagcagag ccaggaaccc ctgtagggaa ggggcaggag agccaggggc
    13261 atgagatggt ggacgagaaa gggggacagg gaagcctgag cgcctctcct gggcttgcca
    13321 aggactcaaa cccagaagcc cagagcaggg ccttagggaa gcgggaccct gctctcggcg
    13381 gaggaatatg tcccagatag cactggggac tctttaagga aagaaggatg gagaaagaga
    13441 aagggggtag aggcagccac gacctggtga acacctagga cgcaccattc tcacaaaggg
    13501 agttttccac acggacatcc ccctcctcac cacagccctg ccaggacggg gctggctact
    13561 ggccttatct cacaggtaaa actgacgcac ggaggaacaa tataaattcg ggactagaaa
    13621 ggtgaagagc caaagttaga actcaggacc aacttattct gattttgttt ttccaaactg
    13681 cttctcctct tgggaagtgt gaggaagctg cagcaccagg atcagtgaaa cgcaccagac
    13741 ggccgcgtca gagcagctca ggttctggga gagggtagcg cagggtggcc actgagaacc
    13801 gggcaggtca cacatccccc acttccctcc caccccctgc caagctctcc ctcccaggat
    13861 ccactctggc tccatcgtaa gcaaacctta gaggttctgg caaggagaga gatggctcca
    13921 ggaaatgggg gtgtgtcacc agataaggaa tctgctcaac aggaggtggg cgttagaccc
    13981 aatatcagga gactaggaag gaggaggcct aaggatgggg cttttctgtc accaatcctg
    14041 tccctagtaa agctgatatc tccaccccac agtgg
//
