LOCUS       sequence-382417-       13691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9-KRAB-MeCP2 in Astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13691)
  AUTHORS   Farrell K, McFadden T, Jarome TJ
  TITLE     Neuronal and astrocytic protein degradation are critical for fear 
            memory formation.
  JOURNAL   Learn Mem. 2023 Mar 15;30(3):70-73. doi: 10.1101/lm.053716.122. 
            Print 2023 Mar.
  PUBMED    36921984
REFERENCE   2  (bases 1 to 13691)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1101/lm.053716.122"; journalName: "Learn Mem"; date: 
            "2023-03-15- 15"; volume: "30"; issue: "3"; pages: "70-73"
COMMENT     Sequence Label: lenti GFAABC1D-dCas9-KRAB-MeCP2
FEATURES             Location/Qualifiers
     source          1..13691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        337..535
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     490..510
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             553..733
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    780..905
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1398..1631
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1816..1860
                     /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    2158..2275
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      3014..3023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3023..3088
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3095..3115
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3140..3163,3164..3166,3167..5653,5654..5656,
                     5657..7240)
                     /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             3140..7240
                     /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             7241..7288
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7349..7543
                     /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             7577..7597
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7607..8464
                     /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    8501..9089
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8554..8574)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8972..8983)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9161..9394
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9420..9437)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9426..9650
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9696..10124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9783..9802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9993..10014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10133..10153)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10138..10467
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10318..10453
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10380..10399
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10515..10562
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10581..10955
                     /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    11085..11206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11122..11141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11176..11195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11255..11271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11255..11271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11268..11290)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11279..11295
                     /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(11303..11309,11310..11327,11328..11333))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11348..11369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11486..11503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11657..12245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11737..11756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12416..13207,13208..13276))
                     /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"
     promoter        complement(13277..13381)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13456..13475)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(13647..13691,1..335)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
       61 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      121 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      181 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      241 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      301 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      361 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      421 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      481 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt
      541 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
      601 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
      661 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
      721 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
      781 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
      841 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
      901 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
      961 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     1021 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     1081 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     1141 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     1201 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     1261 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     1321 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     1381 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     1441 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     1501 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     1561 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     1621 caacagctcc tcgggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     1681 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     1741 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     1801 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     1861 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     1921 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     1981 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     2041 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     2101 aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt
     2161 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     2221 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     2281 ttattacagg gacagcagag atccagtttg gttaattaaa cgcgtaacat atcctggtgt
     2341 ggagtagggg acgctgctct gacagaggct cgggggcctg agctggctct gtgagctggg
     2401 gaggaggcag acagccaggc cttgtctgca agcagacctg gcagcattgg gctggccgcc
     2461 ccccagggcc tcctcttcat gcccagtgaa tgactcacct tggcacagac acaatgttcg
     2521 gggtgggcac agtgcctgct tcccgccgca ccccagcccc cctcaaatgc cttccgagaa
     2581 gcccattgag cagggggctt gcattgcacc ccagcctgac agcctggcat cttgggataa
     2641 aagcagcaca gccccctagg ggctgccctt gctgtgtggc gccaccggcg gtggagaaca
     2701 aggctctatt cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc atggacagtg
     2761 ggtggcaggg ggggagagga gggctgtctg cttcccagaa gtccaaggac acaaatgggt
     2821 gaggggagag ctctccccat agctgggctg cggcccaacc ccaccccctc aggctatgcc
     2881 agggggtgtt gccaggggca cccgggcatc gccagtctag cccactcctt cataaagccc
     2941 tcgcatccca ggagcgagca gagccagagc aggttggaga ggagacgcat cacctccgct
     3001 gctcgcgacc ggtgccacca tggactataa ggaccacgac ggagactaca aggatcatga
     3061 tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat
     3121 ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggccatcg gcaccaactc
     3181 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
     3241 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
     3301 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
     3361 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
     3421 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
     3481 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
     3541 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
     3601 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     3661 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     3721 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     3781 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     3841 tggcctgttc ggcaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     3901 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     3961 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     4021 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     4081 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     4141 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     4201 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
     4261 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     4321 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     4381 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     4441 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     4501 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     4561 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
     4621 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     4681 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     4741 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
     4801 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     4861 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     4921 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     4981 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
     5041 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
     5101 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
     5161 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
     5221 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
     5281 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     5341 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     5401 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     5461 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     5521 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     5581 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     5641 ctacgatgtg gacgctatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa
     5701 ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
     5761 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
     5821 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     5881 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     5941 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
     6001 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     6061 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     6121 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
     6181 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     6241 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
     6301 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
     6361 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
     6421 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
     6481 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
     6541 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
     6601 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
     6661 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
     6721 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
     6781 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
     6841 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
     6901 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
     6961 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
     7021 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
     7081 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
     7141 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
     7201 gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg cggccacgaa
     7261 aaaggccgga caggccaaaa agaaaaagag tggtggagga agtggcgggt cagggtcgat
     7321 ggacgcgaaa tcacttacgg catggtcgag aacactggtt acgttcaagg acgtgtttgt
     7381 ggactttaca cgtgaggagt ggaaattgct ggatactgcg caacaaattg tgtatcgaaa
     7441 tgtcatgctt gagaattaca agaacctcgt cagtctcgga taccagttga cgaaaccgga
     7501 tgtgatcctt aggctcgaaa agggggaaga accttggctg gtatcgggag gtggttcggg
     7561 tggctctgga tcaagcccaa agaagaaacg gaaggtggaa gcctcagtgc aggtgaaaag
     7621 ggtgctggaa aaatcccccg gcaaactcct cgtgaagatg cccttccagg cttcccctgg
     7681 cggaaaaggt gaagggggtg gcgcaaccac atctgcccag gtcatggtca tcaagcgacc
     7741 tggaaggaaa agaaaggccg aggctgaccc tcaggccatt ccaaagaaac ggggacgcaa
     7801 gccagggtcc gtggtcgcag ctgcagcagc tgaggctaag aaaaaggcag tgaaggaaag
     7861 ctccatccgc agtgtgcagg agactgtcct gcccatcaag aagaggaaga ctagggagac
     7921 cgtgtccatc gaggtcaaag aagtggtcaa gcccctgctc gtgtccaccc tgggcgaaaa
     7981 atctggaaag gggctcaaaa catgcaagtc acctggacgg aaaagcaagg agtctagtcc
     8041 aaaggggcgc tcaagctccg cttctagtcc ccctaaaaag gaacaccatc accatcacca
     8101 tcacgccgag tctcctaagg ctcctatgcc actgctccca ccacctccac cacctgagcc
     8161 acagtcaagc gaagacccca tcagcccacc cgagcctcag gatctgtcct ctagtatttg
     8221 caaagaggaa aagatgccca gagcaggcag cctggagagt gatggctgtc caaaagaacc
     8281 cgccaagacc cagcctatgg tggcagccgc tgcaactacc accacaacca caactaccac
     8341 agtggccgaa aaatacaagc atcgcggcga gggcgaacga aaggacattg tgtcaagctc
     8401 catgcccaga cctaaccggg aggaaccagt cgatagtagg acacccgtga ctgagagagt
     8461 ctcataggaa ttcaattcta actagacgcg ttaagtcgac aatcaacctc tggattacaa
     8521 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     8581 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     8641 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     8701 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     8761 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     8821 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     8881 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     8941 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     9001 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     9061 tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca
     9121 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     9181 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     9241 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     9301 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     9361 agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc
     9421 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     9481 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     9541 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     9601 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     9661 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     9721 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     9781 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     9841 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     9901 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     9961 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
    10021 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
    10081 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
    10141 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    10201 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
    10261 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
    10321 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
    10381 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
    10441 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
    10501 tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
    10561 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
    10621 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
    10681 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
    10741 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
    10801 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
    10861 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
    10921 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
    10981 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
    11041 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
    11101 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
    11161 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
    11221 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
    11281 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
    11341 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
    11401 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
    11461 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
    11521 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
    11581 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
    11641 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
    11701 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
    11761 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
    11821 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
    11881 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
    11941 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
    12001 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
    12061 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
    12121 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
    12181 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
    12241 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
    12301 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
    12361 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
    12421 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
    12481 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
    12541 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
    12601 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
    12661 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
    12721 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
    12781 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    12841 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    12901 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    12961 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    13021 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    13081 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    13141 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    13201 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    13261 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    13321 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
    13381 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
    13441 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
    13501 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
    13561 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
    13621 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
    13681 aattacgggg t
//
