LOCUS       sequence-471208-       11309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of SpCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11309)
  AUTHORS   Benedetti L, Fan R, Weigel AV, Moore AS, Houlihan PR, Kittisopikul 
            M, Park G, Petruncio A, Hubbard PM, Pang S, Xu CS, Hess HF, Saalfeld
            S, Rangaraju V, Clapham DE, De Camilli P, Ryan TA, 
            Lippincott-Schwartz J
  TITLE     Periodic ER-plasma membrane junctions support long-range Ca(2+) 
            signal integration in dendrites.
  JOURNAL   Cell. 2025 Jan 23;188(2):484-500.e22. doi: 
            10.1016/j.cell.2024.11.029. Epub 2024 Dec 20.
  PUBMED    39708809
REFERENCE   2  (bases 1 to 11309)
  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: 
            "10.1016/j.cell.2024.11.029"; journalName: "Cell"; date: 
            "2025-01-23- 23"; volume: "188"; issue: "2"; pages: "484-500.e22"
COMMENT     Sequence Label: pLentiSIV hSyn HA-NLS-SpCas9-NLS WPRE
FEATURES             Location/Qualifiers
     source          1..11309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2694..3141
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             3173..3199
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3203..3223
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3233..7333
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3233..7333
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7337..7357
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    7399..7987
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7452..7472)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7870..7881)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8330..8349)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8395..8450
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8449..8468)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8811..8827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8811..8827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8824..8846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8835..8851
                     /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(8859..8865,8866..8883,8884..8889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8904..8925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8984..9179
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9030..9165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    9185..9319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9222..9241)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9276..9295
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9387..9404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9558..10146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9638..9657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10317..11108,11109..11177))
                     /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     10940..10959
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11178..11282)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgatcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacccttctt aacatggctg acaagaagga aactagatga gacagcaggg actttccaca
      421 aggggtgtta cggggaggta ctggggagga gccggtcggg aacacccact ttcttgatgt
      481 ataaatatca ctgcattttg ctatgtagtc agtcgctctg cggagaggct ggcagattga
      541 gccctgggag gttctctcca gcactagcag gtagagcctg ggtgttccct gctagactct
      601 caccagcact tggccggtgc tgggcagagt gactccacgc ttgcttgctt aaagccctct
      661 tcaataaagc tgccatttta gaagtaagct agtgtgtgtt cccatctctc ctagtcgccg
      721 cctggtcaac tcggtactcg gtaataagaa gaccctggtc tgttaggacc ctttctgctt
      781 tgggaaaccg aagcaggaaa atccctagca gattggcgcc cgaacaggga cttgaaggag
      841 agtgagagac tcctgagtac ggctgagtga aggcagtaag ggcggcagga accaaccacg
      901 acggagtgct cctataaagg cgcgggtcgg taccagacgg cgtgaggagc gggagaggaa
      961 gaggcctccg gttgcaggta agtgcaacac aaaaaagaaa tagctgtctt ttatccagga
     1021 agggataata agatagagtg ggagatgggc gcgagaaact ccgtcttgtc agggaagaaa
     1081 gcagatgaat tagaaaaaat taggctacga cccaacggaa agaaaaagta catgttgaag
     1141 catgtagtat gggcagcaaa tgaattagat agatttggat tagcagaaag cctgttggag
     1201 aacaaagaag gatgtcaaaa aatactttcg gtcttagctc cattagtgcc aacaggctca
     1261 gaaaatttaa aaagccttta taatactgtc tgcgtcatct ggtgcattca cgcagaagag
     1321 aaagtgaaac acactgagga agcaaaacag atagtacaga gacacagagg aagagcacca
     1381 tacaatccac agagtcaggg agtagtggaa gcaatgaatc accacctgaa aaatcaaata
     1441 gatagaatca gggaacaagc aaattcagta gaaaccatag tattaatggc agttcattgc
     1501 atgaatttta aaagaagggg aggaataggg gatatgactc cagcagaaag attaattaac
     1561 atgatcacta cagaacaaga aatacaattt caacaatcaa aaaactcaaa atttaaaaat
     1621 tttcgggtct attacagaga aggcagagat caactgtgga agggacccgg tgagctattg
     1681 tggaaagggg aaggagcagt catcttaaag gtagggacag acattaaggt agtacccaga
     1741 agaaaggcta aaattatcaa agattatgga ggaggaaaag agatggatag cagttcccac
     1801 atggaggata ccggagaggc tagagaggtg gcatagcctc atacaattcg attggaattg
     1861 ggagattata aattagtaga gatcactcca attggcttgg cccccacaaa tgtgaggagg
     1921 tacactactg gtggcacctc aagaaataaa agaggggtct ttgtgctagg gttcttgggt
     1981 tttctcgcaa cggcaggttc tgcaatgggc gcggcgtcgt tgacgctgac cgctcagtcc
     2041 cggactttat tggctgggat agtgcagcaa cagcaactgc tgttggacgt ggtcaagaga
     2101 caacaagaat tgttgcgact gaccgtctgg ggaacaaaga acctccagac tagggtcact
     2161 gccatcgaga aatacttaaa ggaccaggcg cagctgaatg cttggggatg tgcgtttaga
     2221 caagtctgcc acactactgt accatggcca aatgcaagtc taacaccaaa gtggaacaat
     2281 gagacttggc aagagtggga gcgaaaggtt gacttcttgg aggaaaatat aacagccctc
     2341 ctagaggagg cacaaattca acaagagaag aacatgtatg aattacaaaa gttaaatagc
     2401 tgggatgtgt ttggcaattg gtttgacctt gcttcttgga taaagtatat acaatatgga
     2461 gtttatatag ttgtaggagt aatactgtta agaatagtaa tctatatagt acaaatgcta
     2521 gctaagttaa ggcaggggta taggccagtg ttctcttccc caccctctta tttccagtag
     2581 acccatatcc aacaggacca ggcactgcca accagagaag gcaaagaagg agacggtgga
     2641 gaaggcggtg gttcgaacgc gccgtttcta gactgcagag ggccctgcgt atgagtgcaa
     2701 gtgggtttta ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac
     2761 ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg
     2821 aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg
     2881 ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg
     2941 tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc
     3001 gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat
     3061 ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc
     3121 gtgtcgtgcc tgagagcgca gtcgagaaac ccgggaaccg gtcgccacca tgtacccata
     3181 cgatgttcca gattacgctt cgccgaagaa aaagcgcaag gtcgaagcgt ccgacaagaa
     3241 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
     3301 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
     3361 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
     3421 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
     3481 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
     3541 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
     3601 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     3661 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     3721 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     3781 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     3841 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     3901 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
     3961 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     4021 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     4081 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     4141 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     4201 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     4261 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
     4321 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     4381 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     4441 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     4501 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     4561 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     4621 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
     4681 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
     4741 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
     4801 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
     4861 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
     4921 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
     4981 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
     5041 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
     5101 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
     5161 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
     5221 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
     5281 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
     5341 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
     5401 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
     5461 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
     5521 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
     5581 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
     5641 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
     5701 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
     5761 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
     5821 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
     5881 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
     5941 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
     6001 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
     6061 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
     6121 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
     6181 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
     6241 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
     6301 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
     6361 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
     6421 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
     6481 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
     6541 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
     6601 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
     6661 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
     6721 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
     6781 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
     6841 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
     6901 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
     6961 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
     7021 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
     7081 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
     7141 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
     7201 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
     7261 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
     7321 gctgggaggc gacagcccca agaagaagag aaaggtggag gccagctaat gtacaaagct
     7381 tcgaattctg cagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     7441 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     7501 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     7561 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     7621 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     7681 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     7741 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     7801 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     7861 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     7921 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     7981 tccccgctga ggccggccgc ggccgcatcg atgccgtata cggtacccct aagaacaatg
     8041 agttacaaat tggcaataga catgtctcat tttataaaag aaaagggggg actggaaggg
     8101 atttattata gtgcaagaag acaagatctg cattttgcta tgtagtcagt cgctctgcgg
     8161 agaggctggc agattgagcc ctgggaggtt ctctccagca ctagcaggta gagcctgggt
     8221 gttccctgct agactctcac cagcacttgg ccggtgctgg gcagagtgac tccacgcttg
     8281 cttgcttaaa gccctcttca ataaagcttc agctgctcga gctagcagat ctttttccct
     8341 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     8401 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     8461 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     8521 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     8581 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     8641 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     8701 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     8761 ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg
     8821 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     8881 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     8941 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag
     9001 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     9061 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     9121 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     9181 agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     9241 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     9301 atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     9361 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     9421 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     9481 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     9541 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     9601 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     9661 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     9721 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     9781 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     9841 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     9901 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     9961 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
    10021 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    10081 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    10141 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    10201 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    10261 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    10321 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    10381 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    10441 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    10501 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    10561 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    10621 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    10681 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    10741 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    10801 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    10861 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    10921 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    10981 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    11041 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    11101 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    11161 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    11221 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    11281 cgcacatttc cccgaaaagt gccacctgg
//
