LOCUS       sequence-318590-       13451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9-KRAB-MeCP2 fusion driven by human SYN promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13451)
  AUTHORS   Duke CG, Bach SV, Revanna JS, Sultan FA, Southern NT, Davis MN, 
            Carullo NVN, Bauman AJ, Phillips RA 3rd, Day JJ
  TITLE     An Improved CRISPR/dCas9 Interference Tool for Neuronal Gene 
            Suppression.
  JOURNAL   Front Genome Ed. 2020 Sep 15;2:9. doi: 10.3389/fgeed.2020.00009. 
            eCollection 2020.
  PUBMED    34713218
REFERENCE   2  (bases 1 to 13451)
  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: "Front 
            Genome Ed."; date: "2020-09-15"; volume: "2:9. doi"; pages: " 
            10.3389/fgeed.2020.00009. eCollection 2020"
COMMENT     Sequence Label: lenti SYN-dCas9-KRAB-MeCP2
FEATURES             Location/Qualifiers
     source          1..13451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2365..2812
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      2819..2828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2828..2893
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2900..2920
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2945..2968,2969..2971,2972..5458,5459..5461,
                     5462..7045)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2945..7045
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7046..7093
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7154..7348
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             7382..7402
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7412..8269
                     /codon_start=1
                     /product="transcriptional repression domain from rat 
                     methyl-CpG-binding protein 2 (Nan et al., 1997)"
                     /label=MeCP2
                     /translation="VQVKRVLEKSPGKLLVKMPFQASPGGKGEGGGATTSAQVMVIKRP
                     GRKRKAEADPQAIPKKRGRKPGSVVAAAAAEAKKKAVKESSIRSVQETVLPIKKRKTRE
                     TVSIEVKEVVKPLLVSTLGEKSGKGLKTCKSPGRKSKESSPKGRSSSASSPPKKEHHHH
                     HHHAESPKAPMPLLPPPPPPEPQSSEDPISPPEPQDLSSSICKEEKMPRAGSLESDGCP
                     KEPAKTQPMVAAAATTTTTTTTTVAEKYKHRGEGERKDIVSSSMPRPNREEPVDSRTPV
                     TERVS"
     misc_feature    8306..8894
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8359..8379)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8777..8788)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8966..9199
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9225..9242)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9231..9455
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9501..9929
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9588..9607)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9798..9819
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9938..9958)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9943..10272
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10123..10258
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10185..10204
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10320..10367
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10386..10760
                     /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    10890..11011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10927..10946)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10981..11000
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11060..11076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11060..11076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11073..11095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11084..11100
                     /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(11108..11114,11115..11132,11133..11138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11153..11174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11291..11308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11462..12050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11542..11561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12221..13012,13013..13081))
                     /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     12844..12863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13082..13186)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13261..13280)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaagtgca agtgggtttt aggaccagga tgaggcgggg
     2401 tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa cccccattcc
     2461 ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc gaggcgcgtg
     2521 cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca
     2581 ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct
     2641 tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc accacgcgag
     2701 gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct
     2761 gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc agaccggtgc
     2821 caccatggac tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga
     2881 cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc
     2941 agccgacaag aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt
     3001 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     3061 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     3121 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     3181 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     3241 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     3301 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     3361 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     3421 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     3481 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     3541 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     3601 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggcaa
     3661 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     3721 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     3781 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     3841 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     3901 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     3961 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     4021 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     4081 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     4141 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     4201 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     4261 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     4321 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     4381 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     4441 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     4501 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     4561 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     4621 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     4681 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     4741 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     4801 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     4861 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     4921 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     4981 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     5041 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     5101 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     5161 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     5221 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     5281 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     5341 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     5401 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacgc
     5461 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     5521 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     5581 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     5641 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     5701 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     5761 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     5821 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     5881 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     5941 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     6001 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     6061 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     6121 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     6181 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     6241 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     6301 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     6361 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     6421 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     6481 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     6541 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     6601 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     6661 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     6721 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     6781 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     6841 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     6901 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     6961 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     7021 cgacctgtct cagctgggag gcgacaaaag gccggcggcc acgaaaaagg ccggacaggc
     7081 caaaaagaaa aagagtggtg gaggaagtgg cgggtcaggg tcgatggacg cgaaatcact
     7141 tacggcatgg tcgagaacac tggttacgtt caaggacgtg tttgtggact ttacacgtga
     7201 ggagtggaaa ttgctggata ctgcgcaaca aattgtgtat cgaaatgtca tgcttgagaa
     7261 ttacaagaac ctcgtcagtc tcggatacca gttgacgaaa ccggatgtga tccttaggct
     7321 cgaaaagggg gaagaacctt ggctggtatc gggaggtggt tcgggtggct ctggatcaag
     7381 cccaaagaag aaacggaagg tggaagcctc agtgcaggtg aaaagggtgc tggaaaaatc
     7441 ccccggcaaa ctcctcgtga agatgccctt ccaggcttcc cctggcggaa aaggtgaagg
     7501 gggtggcgca accacatctg cccaggtcat ggtcatcaag cgacctggaa ggaaaagaaa
     7561 ggccgaggct gaccctcagg ccattccaaa gaaacgggga cgcaagccag ggtccgtggt
     7621 cgcagctgca gcagctgagg ctaagaaaaa ggcagtgaag gaaagctcca tccgcagtgt
     7681 gcaggagact gtcctgccca tcaagaagag gaagactagg gagaccgtgt ccatcgaggt
     7741 caaagaagtg gtcaagcccc tgctcgtgtc caccctgggc gaaaaatctg gaaaggggct
     7801 caaaacatgc aagtcacctg gacggaaaag caaggagtct agtccaaagg ggcgctcaag
     7861 ctccgcttct agtcccccta aaaaggaaca ccatcaccat caccatcacg ccgagtctcc
     7921 taaggctcct atgccactgc tcccaccacc tccaccacct gagccacagt caagcgaaga
     7981 ccccatcagc ccacccgagc ctcaggatct gtcctctagt atttgcaaag aggaaaagat
     8041 gcccagagca ggcagcctgg agagtgatgg ctgtccaaaa gaacccgcca agacccagcc
     8101 tatggtggca gccgctgcaa ctaccaccac aaccacaact accacagtgg ccgaaaaata
     8161 caagcatcgc ggcgagggcg aacgaaagga cattgtgtca agctccatgc ccagacctaa
     8221 ccgggaggaa ccagtcgata gtaggacacc cgtgactgag agagtctcat aggaattcaa
     8281 ttctaactag acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     8341 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     8401 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     8461 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     8521 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     8581 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     8641 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     8701 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     8761 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     8821 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     8881 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     8941 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     9001 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     9061 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     9121 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     9181 gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     9241 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     9301 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     9361 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     9421 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     9481 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     9541 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     9601 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     9661 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     9721 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     9781 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     9841 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     9901 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     9961 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
    10021 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
    10081 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
    10141 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
    10201 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
    10261 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
    10321 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
    10381 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
    10441 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
    10501 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
    10561 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
    10621 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
    10681 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
    10741 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
    10801 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
    10861 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
    10921 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
    10981 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
    11041 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
    11101 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
    11161 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
    11221 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
    11281 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    11341 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    11401 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    11461 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    11521 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    11581 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    11641 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    11701 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    11761 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    11821 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    11881 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
    11941 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    12001 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    12061 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    12121 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
    12181 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
    12241 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
    12301 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
    12361 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
    12421 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    12481 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
    12541 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    12601 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    12661 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    12721 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    12781 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    12841 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    12901 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    12961 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    13021 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    13081 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
    13141 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
    13201 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
    13261 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
    13321 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
    13381 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
    13441 atatacgcgt t
//
