LOCUS       sequence-150192-       14828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dead Cas9 (dCas9) fused to inactive DNMT3A catalytic 
            domain (CD) linked to IRES-mCherry under the control of TetOp and a 
            minimal CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14828)
  AUTHORS   McDonald JI, Celik H, Rois LE, Fishberger G, Fowler T, Rees R, 
            Kramer A, Martens A, Edwards JR, Challen GA
  TITLE     Reprogrammable CRISPR/Cas9-based system for inducing site-specific 
            DNA methylation.
  JOURNAL   Biol Open. 2016 May 11. pii: bio.019067. doi: 10.1242/bio.019067.
  PUBMED    27170255
REFERENCE   2  (bases 1 to 14828)
  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: "Biol Open. 
            2016 May 11. pii: bio.019067. doi: 10.1242/bio.019067."
COMMENT     Sequence Label: TetO-dCas9-mD3A
FEATURES             Location/Qualifiers
     source          1..14828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        455..653
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             671..851
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    898..1023
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1516..1749
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1934..1978
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2276..2393
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2446..2464,2465..2487,2488..2506,2507..2529,2530..2548,
                     2549..2571,2572..2590,2591..2613,2614..2632,2633..2655,
                     2656..2674,2675..2697,2698..2716)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2749..2786
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2783..2807
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2907..2916
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2913..2939,2940..2942,2943..5426,5427..5432,5433..5498,
                     5499..5501,5502..7016)
                     /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
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEIAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2913..7016
                     /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
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEIAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7029..7049
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7059..7082
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7101..8033
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFANVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     misc_feature    8061..8610
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8205..8222)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8432..8451
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8613..9323
                     /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(8758..8776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9026..9045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    9356..9944
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9409..9429)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9827..9838)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9947..9963)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9948..9964)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10469..10649
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10675..10692)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10681..10905
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10951..11379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11038..11057)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11248..11269
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11388..11408)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11393..11722
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11573..11708
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11635..11654
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11770..11817
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11836..12210
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    12340..12461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12377..12396)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12431..12450
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12510..12526)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12510..12526)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12523..12545)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12534..12550
                     /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(12558..12564,12565..12582,12583..12588))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12603..12624
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12741..12758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12912..13500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12992..13011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13671..14462,14463..14531))
                     /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     14294..14313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14532..14636)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14711..14730)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
       61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt
      661 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
      721 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
      781 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
      841 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
      901 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
      961 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     1021 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     1081 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     1141 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     1201 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     1261 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     1321 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     1381 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     1441 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     1501 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     1561 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     1621 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     1681 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     1741 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     1801 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     1861 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     1921 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     1981 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     2041 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     2101 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     2161 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     2221 cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta
     2281 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     2341 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     2401 attacaggga cagcagagat ccagtttggt tatcgagttt accactccct atcagtgata
     2461 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
     2521 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     2581 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     2641 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     2701 ctatcagtga tagagaaaag tgaaagtcga gctcggtacc cgggtcgagt aggcgtgtac
     2761 ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
     2821 atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc cgcggccccg
     2881 aattcgccat acattctaga ggatccgcca ccatggacaa gaagtactcc attgggctcg
     2941 ctatcggcac aaacagcgtc ggctgggccg tcattacgga cgagtacaag gtgccgagca
     3001 aaaaattcaa agttctgggc aataccgatc gccacagcat aaagaagaac ctcattggcg
     3061 ccctcctgtt cgactccggg gagacggccg aagccacgcg gctcaaaaga acagcacggc
     3121 gcagatatac ccgcagaaag aatcggatct gctacctgca ggagatcttt agtaatgaga
     3181 tagctaaggt ggatgactct ttcttccata ggctggagga gtcctttttg gtggaggagg
     3241 ataaaaagca cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg gcgtaccatg
     3301 aaaagtaccc aaccatatat catctgagga agaagcttgt agacagtact gataaggctg
     3361 acttgcggtt gatctatctc gcgctggcgc atatgatcaa atttcgggga cacttcctca
     3421 tcgaggggga cctgaaccca gacaacagcg atgtcgacaa actctttatc caactggttc
     3481 agacttacaa tcagcttttc gaagagaacc cgatcaacgc atccggagtt gacgccaaag
     3541 caatcctgag cgctaggctg tccaaatccc ggcggctcga aaacctcatc gcacagctcc
     3601 ctggggagaa gaagaacggc ctgtttggta atcttatcgc cctgtcactc gggctgaccc
     3661 ccaactttaa atctaacttc gacctggccg aagatgccaa gcttcaactg agcaaagaca
     3721 cctacgatga tgatctcgac aatctgctgg cccagatcgg cgaccagtac gcagaccttt
     3781 ttttggcggc aaagaacctg tcagacgcca ttctgctgag tgatattctg cgagtgaaca
     3841 cggagatcac caaagctccg ctgagcgcta gtatgatcaa gcgctatgat gagcaccacc
     3901 aagacttgac tttgctgaag gcccttgtca gacagcaact gcctgagaag tacaaggaaa
     3961 ttttcttcga tcagtctaaa aatggctacg ccggatacat tgacggcgga gcaagccagg
     4021 aggaatttta caaatttatt aagcccatct tggaaaaaat ggacggcacc gaggagctgc
     4081 tggtaaagct taacagagaa gatctgttgc gcaaacagcg cactttcgac aatggaagca
     4141 tcccccacca gattcacctg ggcgaactgc acgctatcct caggcggcaa gaggatttct
     4201 accccttttt gaaagataac agggaaaaga ttgagaaaat cctcacattt cggataccct
     4261 actatgtagg ccccctcgcc cggggaaatt ccagattcgc gtggatgact cgcaaatcag
     4321 aagagaccat cactccctgg aacttcgagg aagtcgtgga taagggggcc tctgcccagt
     4381 ccttcatcga aaggatgact aactttgata aaaatctgcc taacgaaaag gtgcttccta
     4441 aacactctct gctgtacgag tacttcacag tttataacga gctcaccaag gtcaaatacg
     4501 tcacagaagg gatgagaaag ccagcattcc tgtctggaga gcagaagaaa gctatcgtgg
     4561 acctcctctt caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa gactatttca
     4621 aaaagattga atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc ttcaacgcat
     4681 ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga caaggacttc ctggacaatg
     4741 aggagaacga ggacattctt gaggacattg tcctcaccct tacgttgttt gaagataggg
     4801 agatgattga agaacgcttg aaaacttacg ctcatctctt cgacgacaaa gtcatgaaac
     4861 agctcaagag gcgccgatat acaggatggg ggcggctgtc aagaaaactg atcaatggga
     4921 tccgagacaa gcagagtgga aagacaatcc tggattttct taagtccgat ggatttgcca
     4981 accggaactt catgcagttg atccatgatg actctctcac ctttaaggag gacatccaga
     5041 aagcacaagt ttctggccag ggggacagtc ttcacgagca catcgctaat cttgcaggta
     5101 gcccagctat caaaaaggga atactgcaga ccgttaaggt cgtggatgaa ctcgtcaaag
     5161 taatgggaag gcataagccc gagaatatcg ttatcgagat ggcccgagag aaccaaacta
     5221 cccagaaggg acagaagaac agtagggaaa ggatgaagag gattgaagag ggtataaaag
     5281 aactggggtc ccaaatcctt aaggaacacc cagttgaaaa cacccagctt cagaatgaga
     5341 agctctacct gtactacctg cagaacggca gggacatgta cgtggatcag gaactggaca
     5401 tcaatcggct ctccgactac gacgtggatg ctatcgtgcc ccagtctttt ctcaaagatg
     5461 attctattga taataaagtg ttgacaagat ccgataaaaa tagagggaag agtgataacg
     5521 tcccctcaga agaagttgtc aagaaaatga aaaattattg gcggcagctg ctgaacgcca
     5581 aactgatcac acaacggaag ttcgataatc tgactaaggc tgaacgaggt ggcctgtctg
     5641 agttggataa agccggcttc atcaaaaggc agcttgttga gacacgccag atcaccaagc
     5701 acgtggccca aattctcgat tcacgcatga acaccaagta cgatgaaaat gacaaactga
     5761 ttcgagaggt gaaagttatt actctgaagt ctaagctggt ctcagatttc agaaaggact
     5821 ttcagtttta taaggtgaga gagatcaaca attaccacca tgcgcatgat gcctacctga
     5881 atgcagtggt aggcactgca cttatcaaaa aatatcccaa gcttgaatct gaatttgttt
     5941 acggagacta taaagtgtac gatgttagga aaatgatcgc aaagtctgag caggaaatag
     6001 gcaaggccac cgctaagtac ttcttttaca gcaatattat gaattttttc aagaccgaga
     6061 ttacactggc caatggagag attcggaagc gaccacttat cgaaacaaac ggagaaacag
     6121 gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc ctgtccatgc
     6181 cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg aggcttctcc aaggaaagta
     6241 tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa aaaagattgg gaccccaaga
     6301 aatacggcgg attcgattct cctacagtcg cttacagtgt actggttgtg gccaaagtgg
     6361 agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc acaatcatgg
     6421 agcgatcaag cttcgaaaaa aaccccatcg actttctcga ggcgaaagga tataaagagg
     6481 tcaaaaaaga cctcatcatt aagcttccca agtactctct ctttgagctt gaaaacggcc
     6541 ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg taacgagctg gcactgccct
     6601 ctaaatacgt taatttcttg tatctggcca gccactatga aaagctcaaa gggtctcccg
     6661 aagataatga gcagaagcag ctgttcgtgg aacaacacaa acactacctt gatgagatca
     6721 tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc cgacgctaac ctcgataagg
     6781 tgctttctgc ttacaataag cacagggata agcccatcag ggagcaggca gaaaacatta
     6841 tccacttgtt tactctgacc aacttgggcg cgcctgcagc cttcaagtac ttcgacacca
     6901 ccatagacag aaagcggtac acctctacaa aggaggtcct ggacgccaca ctgattcatc
     6961 agtcaattac ggggctctat gaaacaagaa tcgacctctc tcagctcggt ggagacagca
     7021 gggctgaccc caagaagaag aggaaggtgg gagctagcga ctacaaagac gatgacgaca
     7081 agggaagccg agaggactgg ccctcccggc tccagatgtt cttcgctaat aaccacgacc
     7141 aggaatttga ccctccaaag gtttacccac ctgtcccagc tgagaagagg aagcccatcc
     7201 gggtgctgtc tctctttgat ggaatcgcta cagggctcct ggtgctgaag gacttgggca
     7261 ttcaggtgga ccgctacatt gcctcggagg tgtgtgagga ctccatcacg gtgggcatgg
     7321 tgcggcacca ggggaagatc atgtacgtcg gggacgtccg cagcgtcaca cagaagcata
     7381 tccaggagtg gggcccattc gatctggtga ttgggggcag tccctgcaat gacctctcca
     7441 tcgtcaaccc tgctcgcaag ggcctctacg agggcactgg ccggctcttc tttgagttct
     7501 accgcctcct gcatgatgcg cggcccaagg agggagatga tcgccccttc ttctggctct
     7561 ttgcgaatgt ggtggccatg ggcgttagtg acaagaggga catctcgcga tttctcgagt
     7621 ccaaccctgt gatgattgat gccaaagaag tgtcagctgc acacagggcc cgctacttct
     7681 ggggtaacct tcccggtatg aacaggccgt tggcatccac tgtgaatgat aagctggagc
     7741 tgcaggagtg tctggagcat ggcaggatag ccaagttcag caaagtgagg accattacta
     7801 cgaggtcaaa ctccataaag cagggcaaag accagcattt tcctgtcttc atgaatgaga
     7861 aagaggacat cttatggtgc actgaaatgg aaagggtatt tggtttccca gtccactata
     7921 ctgacgtctc caacatgagc cgcttggcga ggcagagact gctgggccgg tcatggagcg
     7981 tgccagtcat ccgccacctc ttcgctccgc tgaaggagta ttttgcgtgt gtgtaaaccg
     8041 gtactcgacc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     8101 gcttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     8161 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     8221 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     8281 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcccctg
     8341 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     8401 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     8461 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     8521 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     8581 cgtggttttc ctttgaaaaa cacgataata ccatggtgag caagggcgag gaggataaca
     8641 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     8701 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     8761 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     8821 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     8881 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     8941 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     9001 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     9061 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     9121 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     9181 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     9241 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     9301 gcatggacga gctgtacaag tagacgcgcc cgaattcgat atcaagctta tcgataatca
     9361 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     9421 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     9481 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     9541 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     9601 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     9661 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     9721 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     9781 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     9841 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     9901 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     9961 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
    10021 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
    10081 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
    10141 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
    10201 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
    10261 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
    10321 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
    10381 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
    10441 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
    10501 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
    10561 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
    10621 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
    10681 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
    10741 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
    10801 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
    10861 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
    10921 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
    10981 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
    11041 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
    11101 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
    11161 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    11221 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
    11281 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
    11341 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
    11401 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
    11461 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
    11521 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
    11581 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
    11641 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
    11701 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
    11761 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
    11821 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
    11881 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
    11941 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
    12001 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
    12061 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
    12121 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
    12181 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
    12241 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
    12301 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
    12361 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
    12421 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
    12481 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
    12541 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
    12601 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
    12661 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
    12721 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
    12781 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
    12841 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
    12901 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
    12961 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
    13021 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
    13081 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
    13141 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
    13201 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
    13261 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
    13321 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
    13381 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
    13441 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
    13501 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
    13561 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
    13621 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
    13681 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
    13741 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
    13801 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
    13861 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
    13921 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
    13981 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
    14041 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    14101 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    14161 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    14221 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    14281 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    14341 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    14401 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    14461 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    14521 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    14581 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    14641 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
    14701 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
    14761 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
    14821 tgaccgac
//
