LOCUS       sequence-211273-       10147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KRAB fused to N-terminus of dCas9; pCDNA3 vector backbone, mammalian
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10147)
  AUTHORS   O'Geen H, Ren C, Nicolet CM, Perez AA, Halmai J, Le VM, Mackay JP, 
            Farnham PJ, Segal DJ
  TITLE     dCas9-based epigenome editing suggests acquisition of histone 
            methylation is not sufficient for target gene repression.
  JOURNAL   Nucleic Acids Res. 2017 Sep 29;45(17):9901-9916. doi: 
            10.1093/nar/gkx578.
  PUBMED    28973434
REFERENCE   2  (bases 1 to 10147)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2017-09-29- 29"; volume: "45"; issue: 
            "17"; pages: "9901-9916"
COMMENT     Sequence Label: KRAB-dCas9
FEATURES             Location/Qualifiers
     source          1..10147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        208..411
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     362..382
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     408..432
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             529..549
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             568..633
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             664..855
                     /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="TLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQ
                     LTKPDVILRLEKGEEPWLV"
     CDS             join(946..972,973..975,976..3459,3460..3465,3466..3531,
                     3532..3534,3535..5049)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /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             join(946..972,973..975,976..3462,3463..3465,3466..5049)
                     /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             946..5049
                     /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             5125..5172
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5191..5247
                     /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"
     primer_bind     complement(5311..5330)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5355..5403
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5605..6033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5692..5711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5902..5923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6042..6062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6047..6376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6227..6362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6289..6308
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6443..7237
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6497..6516)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7107..7126
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7413..7534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7450..7469)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7504..7523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7583..7599)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7583..7599)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7596..7618)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7607..7623
                     /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(7631..7637,7638..7655,7656..7661))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7676..7697
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7814..7831)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7985..8573)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8065..8084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8744..9535,9536..9604))
                     /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     9367..9386
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9605..9709)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9784..9803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(9975..10147,1..207)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
       61 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      121 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      181 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      241 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      301 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      361 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      421 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
      481 gaccgatcca gcctccggac tctagaggat cgaacccttg ccaccatgcc caagaagaag
      541 aggaaggtgg gaggctccgg aggaagcgac tataaggacc acgacggaga ctacaaggat
      601 catgatattg attacaaaga cgatgacgat aagggtggcg ggtccggcgg tggatccggt
      661 accacactgg tgaccttcaa ggatgtattt gtggacttca ccagggagga gtggaagctg
      721 ctggacactg ctcagcagat cgtgtacaga aatgtgatgc tggagaacta taagaacctg
      781 gtttccttgg gttatcagct tactaagcca gatgtgatcc tccggttgga gaagggagaa
      841 gagccctggc tggtggagag agaaattcac caagagaccc atcctagtac cggagggagt
      901 ggtggaagcg gcggttctgg tggctcaggc ggcagcggcc ggccaatgga caagaagtac
      961 tccattgggc tcgctatcgg cacaaacagc gtcggctggg ccgtcattac ggacgagtac
     1021 aaggtgccga gcaaaaaatt caaagttctg ggcaataccg atcgccacag cataaagaag
     1081 aacctcattg gcgccctcct gttcgactcc ggggagacgg ccgaagccac gcggctcaaa
     1141 agaacagcac ggcgcagata tacccgcaga aagaatcgga tctgctacct gcaggagatc
     1201 tttagtaatg agatggctaa ggtggatgac tctttcttcc ataggctgga ggagtccttt
     1261 ttggtggagg aggataaaaa gcacgagcgc cacccaatct ttggcaatat cgtggacgag
     1321 gtggcgtacc atgaaaagta cccaaccata tatcatctga ggaagaagct tgtagacagt
     1381 actgataagg ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat caaatttcgg
     1441 ggacacttcc tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga caaactcttt
     1501 atccaactgg ttcagactta caatcagctt ttcgaagaga acccgatcaa cgcatccgga
     1561 gttgacgcca aagcaatcct gagcgctagg ctgtccaaat cccggcggct cgaaaacctc
     1621 atcgcacagc tccctgggga gaagaagaac ggcctgtttg gtaatcttat cgccctgtca
     1681 ctcgggctga cccccaactt taaatctaac ttcgacctgg ccgaagatgc caagcttcaa
     1741 ctgagcaaag acacctacga tgatgatctc gacaatctgc tggcccagat cggcgaccag
     1801 tacgcagacc tttttttggc ggcaaagaac ctgtcagacg ccattctgct gagtgatatt
     1861 ctgcgagtga acacggagat caccaaagct ccgctgagcg ctagtatgat caagcgctat
     1921 gatgagcacc accaagactt gactttgctg aaggcccttg tcagacagca actgcctgag
     1981 aagtacaagg aaattttctt cgatcagtct aaaaatggct acgccggata cattgacggc
     2041 ggagcaagcc aggaggaatt ttacaaattt attaagccca tcttggaaaa aatggacggc
     2101 accgaggagc tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc
     2161 gacaatggaa gcatccccca ccagattcac ctgggcgaac tgcacgctat cctcaggcgg
     2221 caagaggatt tctacccctt tttgaaagat aacagggaaa agattgagaa aatcctcaca
     2281 tttcggatac cctactatgt aggccccctc gcccggggaa attccagatt cgcgtggatg
     2341 actcgcaaat cagaagagac catcactccc tggaacttcg aggaagtcgt ggataagggg
     2401 gcctctgccc agtccttcat cgaaaggatg actaactttg ataaaaatct gcctaacgaa
     2461 aaggtgcttc ctaaacactc tctgctgtac gagtacttca cagtttataa cgagctcacc
     2521 aaggtcaaat acgtcacaga agggatgaga aagccagcat tcctgtctgg agagcagaag
     2581 aaagctatcg tggacctcct cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa
     2641 gaagactatt tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg agtggaggat
     2701 cgcttcaacg catccctggg aacgtatcac gatctcctga aaatcattaa agacaaggac
     2761 ttcctggaca atgaggagaa cgaggacatt cttgaggaca ttgtcctcac ccttacgttg
     2821 tttgaagata gggagatgat tgaagaacgc ttgaaaactt acgctcatct cttcgacgac
     2881 aaagtcatga aacagctcaa gaggcgccga tatacaggat gggggcggct gtcaagaaaa
     2941 ctgatcaatg ggatccgaga caagcagagt ggaaagacaa tcctggattt tcttaagtcc
     3001 gatggatttg ccaaccggaa cttcatgcag ttgatccatg atgactctct cacctttaag
     3061 gaggacatcc agaaagcaca agtttctggc cagggggaca gtcttcacga gcacatcgct
     3121 aatcttgcag gtagcccagc tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat
     3181 gaactcgtca aagtaatggg aaggcataag cccgagaata tcgttatcga gatggcccga
     3241 gagaaccaaa ctacccagaa gggacagaag aacagtaggg aaaggatgaa gaggattgaa
     3301 gagggtataa aagaactggg gtcccaaatc cttaaggaac acccagttga aaacacccag
     3361 cttcagaatg agaagctcta cctgtactac ctgcagaacg gcagggacat gtacgtggat
     3421 caggaactgg acatcaatcg gctctccgac tacgacgtgg atgctatcgt gccccagtct
     3481 tttctcaaag atgattctat tgataataaa gtgttgacaa gatccgataa aaatagaggg
     3541 aagagtgata acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag
     3601 ctgctgaacg ccaaactgat cacacaacgg aagttcgata atctgactaa ggctgaacga
     3661 ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc
     3721 cagatcacca agcacgtggc ccaaattctc gattcacgca tgaacaccaa gtacgatgaa
     3781 aatgacaaac tgattcgaga ggtgaaagtt attactctga agtctaagct ggtctcagat
     3841 ttcagaaagg actttcagtt ttataaggtg agagagatca acaattacca ccatgcgcat
     3901 gatgcctacc tgaatgcagt ggtaggcact gcacttatca aaaaatatcc caagcttgaa
     3961 tctgaatttg tttacggaga ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct
     4021 gagcaggaaa taggcaaggc caccgctaag tacttctttt acagcaatat tatgaatttt
     4081 ttcaagaccg agattacact ggccaatgga gagattcgga agcgaccact tatcgaaaca
     4141 aacggagaaa caggagaaat cgtgtgggac aagggtaggg atttcgcgac agtccggaag
     4201 gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac cggaggcttc
     4261 tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat
     4321 tgggacccca agaaatacgg cggattcgat tctcctacag tcgcttacag tgtactggtt
     4381 gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc
     4441 atcacaatca tggagcgatc aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa
     4501 ggatataaag aggtcaaaaa agacctcatc attaagcttc ccaagtactc tctctttgag
     4561 cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg agctgcagaa aggtaacgag
     4621 ctggcactgc cctctaaata cgttaatttc ttgtatctgg ccagccacta tgaaaagctc
     4681 aaagggtctc ccgaagataa tgagcagaag cagctgttcg tggaacaaca caaacactac
     4741 cttgatgaga tcatcgagca aataagcgaa ttctccaaaa gagtgatcct cgccgacgct
     4801 aacctcgata aggtgctttc tgcttacaat aagcacaggg ataagcccat cagggagcag
     4861 gcagaaaaca ttatccactt gtttactctg accaacttgg gcgcgcctgc agccttcaag
     4921 tacttcgaca ccaccataga cagaaagcgg tacacctcta caaaggaggt cctggacgcc
     4981 acactgattc atcagtcaat tacggggctc tatgaaacaa gaatcgacct ctctcagctc
     5041 ggtggagacg gtggtagtgg aggttcagga ggatccgggg ggagcggagg gagcgctagc
     5101 ggtggaggct ccggaggtgg atctaaaagg ccggcggcca cgaaaaaggc cggtcaggca
     5161 aaaaagaaaa agggtggtag tggaagcgga gctactaact tcagcctgct gaagcaggct
     5221 ggagacgtgg aggagaaccc tggacctgcg gccgcatgaa agggttcgat ccctaccggt
     5281 tagtaatgag tttaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5341 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     5401 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5461 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5521 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca
     5581 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5641 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5701 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5761 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5821 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5881 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5941 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     6001 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     6061 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6121 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6181 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     6241 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     6301 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     6361 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     6421 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     6481 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     6541 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     6601 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     6661 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     6721 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     6781 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     6841 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     6901 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6961 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     7021 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     7081 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     7141 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     7201 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     7261 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     7321 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     7381 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     7441 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     7501 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     7561 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7621 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7681 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7741 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7801 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7861 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7921 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7981 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8101 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     8161 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     8221 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8281 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     8341 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     8401 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     8461 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8521 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8581 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8641 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8701 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8761 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8821 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8881 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8941 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     9001 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     9061 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     9121 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     9181 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     9241 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     9301 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     9361 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     9421 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     9481 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     9541 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9601 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9661 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9721 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct
     9781 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     9841 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     9901 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     9961 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
    10021 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
    10081 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
    10141 aacgcca
//
