LOCUS       sequence-300738-       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"
     primer_bind     85..104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(323..427)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(502..521)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        693..1072
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1074..1272
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1227..1247
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1290..1470
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1517..1642
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2135..2368
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2553..2597
                     /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    2895..3012
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3057..3504
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3511..3520
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3520..3585
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3592..3612
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3637..3660,3661..3663,3664..6150,6151..6153,
                     6154..7737)
                     /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             3637..7737
                     /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             7738..7785
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7846..8040
                     /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             8074..8094
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8104..8961
                     /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    8998..9586
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9051..9071)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9469..9480)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9658..9891
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9917..9934)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9923..10147
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10193..10621
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10280..10299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10490..10511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10630..10650)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10635..10964
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10815..10950
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10877..10896
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11012..11059
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11078..11452
                     /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    11582..11703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11619..11638)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11673..11692
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11752..11768)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11752..11768)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11765..11787)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11776..11792
                     /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(11800..11806,11807..11824,11825..11830))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11845..11866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11983..12000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12154..12742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12234..12253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12913..13451,1..253,254..322))
                     /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"
ORIGIN
        1 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
       61 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
      121 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
      181 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
      241 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
      301 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
      361 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
      421 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
      481 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
      541 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
      601 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
      661 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      721 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      781 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      841 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      901 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      961 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1021 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     1081 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     1141 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     1201 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     1261 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
     1321 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     1381 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     1441 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     1501 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     1561 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     1621 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     1681 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1741 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1801 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1861 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1921 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1981 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     2041 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     2101 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     2161 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     2221 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     2281 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     2341 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     2401 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     2461 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     2521 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     2581 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     2641 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     2701 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     2761 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     2821 gatccattcg attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca
     2881 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     2941 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     3001 attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggtt aattaaagtg
     3061 caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc
     3121 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg
     3181 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca
     3241 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg
     3301 acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc
     3361 gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac
     3421 catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga
     3481 gtcgtgtcgt gcctgagagc gcagaccggt gccaccatgg actataagga ccacgacgga
     3541 gactacaagg atcatgatat tgattacaaa gacgatgacg ataagatggc cccaaagaag
     3601 aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag catcggcctg
     3661 gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     3721 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     3781 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
     3841 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     3901 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     3961 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     4021 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     4081 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     4141 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     4201 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     4261 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     4321 cccggcgaga agaagaatgg cctgttcggc aacctgattg ccctgagcct gggcctgacc
     4381 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     4441 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     4501 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     4561 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     4621 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     4681 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     4741 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     4801 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     4861 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     4921 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     4981 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     5041 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     5101 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     5161 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     5221 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     5281 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     5341 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     5401 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     5461 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     5521 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     5581 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     5641 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     5701 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     5761 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     5821 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     5881 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     5941 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     6001 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     6061 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     6121 atcaaccggc tgtccgacta cgatgtggac gctatcgtgc ctcagagctt tctgaaggac
     6181 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
     6241 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
     6301 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     6361 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     6421 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
     6481 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     6541 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     6601 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     6661 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     6721 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     6781 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
     6841 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
     6901 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     6961 atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
     7021 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
     7081 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
     7141 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
     7201 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     7261 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
     7321 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     7381 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
     7441 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
     7501 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
     7561 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
     7621 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
     7681 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaaa
     7741 aggccggcgg ccacgaaaaa ggccggacag gccaaaaaga aaaagagtgg tggaggaagt
     7801 ggcgggtcag ggtcgatgga cgcgaaatca cttacggcat ggtcgagaac actggttacg
     7861 ttcaaggacg tgtttgtgga ctttacacgt gaggagtgga aattgctgga tactgcgcaa
     7921 caaattgtgt atcgaaatgt catgcttgag aattacaaga acctcgtcag tctcggatac
     7981 cagttgacga aaccggatgt gatccttagg ctcgaaaagg gggaagaacc ttggctggta
     8041 tcgggaggtg gttcgggtgg ctctggatca agcccaaaga agaaacggaa ggtggaagcc
     8101 tcagtgcagg tgaaaagggt gctggaaaaa tcccccggca aactcctcgt gaagatgccc
     8161 ttccaggctt cccctggcgg aaaaggtgaa gggggtggcg caaccacatc tgcccaggtc
     8221 atggtcatca agcgacctgg aaggaaaaga aaggccgagg ctgaccctca ggccattcca
     8281 aagaaacggg gacgcaagcc agggtccgtg gtcgcagctg cagcagctga ggctaagaaa
     8341 aaggcagtga aggaaagctc catccgcagt gtgcaggaga ctgtcctgcc catcaagaag
     8401 aggaagacta gggagaccgt gtccatcgag gtcaaagaag tggtcaagcc cctgctcgtg
     8461 tccaccctgg gcgaaaaatc tggaaagggg ctcaaaacat gcaagtcacc tggacggaaa
     8521 agcaaggagt ctagtccaaa ggggcgctca agctccgctt ctagtccccc taaaaaggaa
     8581 caccatcacc atcaccatca cgccgagtct cctaaggctc ctatgccact gctcccacca
     8641 cctccaccac ctgagccaca gtcaagcgaa gaccccatca gcccacccga gcctcaggat
     8701 ctgtcctcta gtatttgcaa agaggaaaag atgcccagag caggcagcct ggagagtgat
     8761 ggctgtccaa aagaacccgc caagacccag cctatggtgg cagccgctgc aactaccacc
     8821 acaaccacaa ctaccacagt ggccgaaaaa tacaagcatc gcggcgaggg cgaacgaaag
     8881 gacattgtgt caagctccat gcccagacct aaccgggagg aaccagtcga tagtaggaca
     8941 cccgtgactg agagagtctc ataggaattc aattctaact agacgcgtta agtcgacaat
     9001 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     9061 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     9121 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     9181 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     9241 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     9301 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     9361 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     9421 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     9481 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     9541 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     9601 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     9661 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     9721 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     9781 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     9841 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt
     9901 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     9961 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
    10021 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
    10081 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
    10141 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
    10201 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
    10261 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
    10321 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
    10381 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
    10441 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
    10501 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
    10561 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
    10621 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
    10681 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
    10741 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
    10801 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
    10861 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
    10921 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
    10981 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
    11041 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
    11101 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
    11161 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
    11221 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
    11281 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
    11341 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
    11401 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
    11461 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
    11521 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
    11581 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
    11641 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
    11701 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
    11761 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    11821 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    11881 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    11941 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    12001 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    12061 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    12121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    12181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    12241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    12301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    12361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    12421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    12481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    12541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    12601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    12661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    12721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    12781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    12841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    12901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    12961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    13021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    13081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    13141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    13201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    13261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    13321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    13381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    13441 aagatgcttt t
//
