LOCUS       sequence-369311-       12480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9-24xGCN4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12480)
  AUTHORS   Chaudhary N, Nho SH, Cho H, Gantumur N, Ra JS, Myung K, Kim H
  TITLE     Background-suppressed live visualization of genomic loci with an 
            improved CRISPR system based on a split fluorophore.
  JOURNAL   Genome Res. 2020 Sep;30(9):1306-1316. doi: 10.1101/gr.260018.119. 
            Epub 2020 Sep 4.
  PUBMED    32887690
REFERENCE   2  (bases 1 to 12480)
  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/gr.260018.119"; journalName: "Genome Res"; date: "2020-09";
            volume: "30"; issue: "9"; pages: "1306-1316"
COMMENT     Sequence Label: pM201
FEATURES             Location/Qualifiers
     source          1..12480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2238..2617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2618..2821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2772..2792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2840..2849
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2852..2878
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2885..2905
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2906..2929,2930..2932,2933..5419,5420..5422,
                     5423..7006)
                     /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             7016..7036
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7091..7147
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7163..7219
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7235..7291
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7307..7363
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7379..7435
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7451..7507
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7523..7579
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7595..7651
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7667..7723
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7739..7795
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7811..7867
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7883..7939
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7955..8011
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8027..8083
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8099..8155
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8171..8227
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8243..8299
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8315..8371
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8387..8443
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8459..8515
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8531..8587
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8603..8659
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8747..8803
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             8810..8830
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    8870..9458
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8923..8943)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9341..9352)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9533..9766
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9885..9903)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9971..10075
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10076..10144,10145..10936)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10294..10313)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11107..11695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11596..11615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11849..11866
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12106..12127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12142..12147,12148..12165,12166..12172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12180..12196
                     /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)."
     primer_bind     12185..12207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12204..12220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12204..12220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12270..12289
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttgacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
     2221 gctagactcg aggcgttgac attgattatt gactagttat taatagtaat caattacggg
     2281 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     2341 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     2401 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     2461 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     2521 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     2581 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     2641 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2701 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     2761 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     2821 tctctggcta actaccggtg ccaccatggg atacccatac gatgttccag attacgctgc
     2881 tagccccaaa aagaagagga aagtggacaa gaagtattct atcggactgg ccatcgggac
     2941 taatagcgtc gggtgggccg tgatcactga cgagtacaag gtgccctcta agaagttcaa
     3001 ggtgctcggg aacaccgacc ggcattccat caagaaaaat ctgatcggag ctctcctctt
     3061 tgattcaggg gagaccgctg aagcaacccg cctcaagcgg actgctagac ggcggtacac
     3121 caggaggaag aaccggattt gttaccttca agagatattc tccaacgaaa tggcaaaggt
     3181 cgacgacagc ttcttccata ggctggaaga atcattcctc gtggaagagg ataagaagca
     3241 tgaacggcat cccatcttcg gtaatatcgt cgacgaggtg gcctatcacg agaaataccc
     3301 aaccatctac catcttcgca aaaagctggt ggactcaacc gacaaggcag acctccggct
     3361 tatctacctg gccctggccc acatgatcaa gttcagaggc cacttcctga tcgagggcga
     3421 cctcaatcct gacaatagcg atgtggataa actgttcatc cagctggtgc agacttacaa
     3481 ccagctcttt gaagagaacc ccatcaatgc aagcggagtc gatgccaagg ccattctgtc
     3541 agcccggctg tcaaagagcc gcagacttga gaatcttatc gctcagctgc cgggtgaaaa
     3601 gaaaaatgga ctgttcggga acctgattgc tctttcactt gggctgactc ccaatttcaa
     3661 gtctaatttc gacctggcag aggatgccaa gctgcaactg tccaaggaca cctatgatga
     3721 cgatctcgac aacctcctgg cccagatcgg tgaccaatac gccgaccttt tccttgctgc
     3781 taagaatctt tctgacgcca tcctgctgtc tgacattctc cgcgtgaaca ctgaaatcac
     3841 caaggcccct ctttcagctt caatgattaa gcggtatgat gagcaccacc aggacctgac
     3901 cctgcttaag gcactcgtcc ggcagcagct tccggagaag tacaaggaaa tcttctttga
     3961 ccagtcaaag aatggatacg ccggctacat cgacggaggt gcctcccaag aggaatttta
     4021 taagtttatc aaacctatcc ttgagaagat ggacggcacc gaagagctcc tcgtgaaact
     4081 gaatcgggag gatctgctgc ggaagcagcg cactttcgac aatgggagca ttccccacca
     4141 gatccatctt ggggagcttc acgccatcct tcggcgccaa gaggacttct acccctttct
     4201 taaggacaac agggagaaga ttgagaaaat tctcactttc cgcatcccct actacgtggg
     4261 acccctcgcc agaggaaata gccggtttgc ttggatgacc agaaagtcag aagaaactat
     4321 cactccctgg aacttcgaag aggtggtgga caagggagcc agcgctcagt cattcatcga
     4381 acggatgact aacttcgata agaacctccc caatgagaag gtcctgccga aacattccct
     4441 gctctacgag tactttaccg tgtacaacga gctgaccaag gtgaaatatg tcaccgaagg
     4501 gatgaggaag cccgcattcc tgtcaggcga acaaaagaag gcaattgtgg accttctgtt
     4561 caagaccaat agaaaggtga ccgtgaagca gctgaaggag gactatttca agaaaattga
     4621 atgcttcgac tctgtggaga ttagcggggt cgaagatcgg ttcaacgcaa gcctgggtac
     4681 ctaccatgat ctgcttaaga tcatcaagga caaggatttt ctggacaatg aggagaacga
     4741 ggacatcctt gaggacattg tcctgactct cactctgttc gaggaccggg aaatgatcga
     4801 ggagaggctt aagacctacg cccatctgtt cgacgataaa gtgatgaagc aacttaaacg
     4861 gagaagatat accggatggg gacgccttag ccgcaaactc atcaacggaa tccgggacaa
     4921 acagagcgga aagaccattc ttgatttcct taagagcgac ggattcgcta atcgcaactt
     4981 catgcaactt atccatgatg attccctgac ctttaaggag gacatccaga aggcccaagt
     5041 gtctggacaa ggtgactcac tgcacgagca tatcgcaaat ctggctggtt cacccgctat
     5101 taagaagggt attctccaga ccgtgaaagt cgtggacgag ctggtcaagg tgatgggtcg
     5161 ccataaacca gagaacattg tcatcgagat ggccagggaa aaccagacta cccagaaggg
     5221 acagaagaac agcagggagc ggatgaaaag aattgaggaa gggattaagg agctcgggtc
     5281 acagatcctt aaagagcacc cggtggaaaa cacccagctt cagaatgaga agctctatct
     5341 gtactacctt caaaatggac gcgatatgta tgtggaccaa gagcttgata tcaacaggct
     5401 ctcagactac gacgtggacg ccatcgtccc tcagagcttc ctcaaagacg actcaattga
     5461 caataaggtg ctgactcgct cagacaagaa ccggggaaag tcagataacg tgccctcaga
     5521 ggaagtcgtg aaaaagatga agaactattg gcgccagctt ctgaacgcaa agctgatcac
     5581 tcagcggaag ttcgacaatc tcactaaggc tgagaggggc ggactgagcg aactggacaa
     5641 agcaggattc attaaacggc aacttgtgga gactcggcag attactaaac atgtcgccca
     5701 aatccttgac tcacgcatga ataccaagta cgacgaaaac gacaaactta tccgcgaggt
     5761 gaaggtgatt accctgaagt ccaagctggt cagcgatttc agaaaggact ttcaattcta
     5821 caaagtgcgg gagatcaata actatcatca tgctcatgac gcatatctga atgccgtggt
     5881 gggaaccgcc ctgatcaaga agtacccaaa gctggaaagc gagttcgtgt acggagacta
     5941 caaggtctac gacgtgcgca agatgattgc caaatctgag caggagatcg gaaaggccac
     6001 cgcaaagtac ttcttctaca gcaacatcat gaatttcttc aagaccgaaa tcacccttgc
     6061 aaacggtgag atccggaaga ggccgctcat cgagactaat ggggagactg gcgaaatcgt
     6121 gtgggacaag ggcagagatt tcgctaccgt gcgcaaagtg ctttctatgc ctcaagtgaa
     6181 catcgtgaag aaaaccgagg tgcaaaccgg aggcttttct aaggaatcaa tcctccccaa
     6241 gcgcaactcc gacaagctca ttgcaaggaa gaaggattgg gaccctaaga agtacggcgg
     6301 attcgattca ccaactgtgg cttattctgt cctggtcgtg gctaaggtgg aaaaaggaaa
     6361 gtctaagaag ctcaagagcg tgaaggaact gctgggtatc accattatgg agcgcagctc
     6421 cttcgagaag aacccaattg actttctcga agccaaaggt tacaaggaag tcaagaagga
     6481 ccttatcatc aagctcccaa agtatagcct gttcgaactg gagaatgggc ggaagcggat
     6541 gctcgcctcc gctggcgaac ttcagaaggg taatgagctg gctctcccct ccaagtacgt
     6601 gaatttcctc taccttgcaa gccattacga gaagctgaag gggagccccg aggacaacga
     6661 gcaaaagcaa ctgtttgtgg agcagcataa gcattatctg gacgagatca ttgagcagat
     6721 ttccgagttt tctaaacgcg tcattctcgc tgatgccaac ctcgataaag tccttagcgc
     6781 atacaataag cacagagaca aaccaattcg ggagcaggct gagaatatca tccacctgtt
     6841 caccctcacc aatcttggtg cccctgccgc attcaagtac ttcgacacca ccatcgaccg
     6901 gaaacgctat acctccacca aagaagtgct ggacgccacc ctcatccacc agagcatcac
     6961 cggactttac gaaactcgga ttgacctctc acagctcgga ggggatgagg gagctcccaa
     7021 gaaaaagcgc aaggtaggta gttccggatc caacggtccg ggtggatctg gaggtggagg
     7081 ttctggagga gaagaacttt tgagcaagaa ttatcatctt gagaacgaag tggctcgtct
     7141 taagaaaggt tctggcagtg gagaagaact gctttcaaag aattaccacc tggaaaatga
     7201 ggtagctaga ctgaaaaagg ggagcggaag tggggaggag ttgctgagca aaaattatca
     7261 tttggagaac gaagtagcac gactaaagaa agggtccgga tcgggtgagg agttactctc
     7321 gaaaaattat catctcgaaa acgaagtggc tcggctaaaa aagggcagtg gttctggaga
     7381 agagctatta tctaaaaact accacctcga aaatgaggtg gcacgcttaa aaaagggaag
     7441 tggcagtggt gaagagctac tatccaagaa ttatcatctt gagaacgagg tagcgcgttt
     7501 gaagaagggt tccggctcag gagaggaact gctctcgaag aactatcatc ttgaaaatga
     7561 ggtcgctcga ttaaaaaagg gatcgggcag tggtgaggaa ctactttcaa agaattacca
     7621 cctcgaaaac gaagtagctc gattaaagaa aggttcaggg tcgggtgaag aattactgag
     7681 taaaaattat catctggaaa atgaggtagc gagactaaaa aaggggagtg gttctggcga
     7741 agagttgcta tcgaaaaatt atcatcttga gaacgaagtt gctaggctca aaaagggctc
     7801 aggctcaggc gaggagttgc tctcgaaaaa ctaccacttg gaaaatgagg tcgcgaggtt
     7861 gaaaaagggg agcgggtcgg gcgaggagtt attgagcaaa aactatcatt tagagaacga
     7921 agtcgcgcgc ttaaagaaag gctcgggctc gggcgaagaa ctcttatcga agaactacca
     7981 cctcgaaaat gaggtcgcca ggttgaaaaa gggcagtggc agcggggagg aactcttgag
     8041 caagaactac cacttggaga atgaggtcgc gagattgaag aaagggtcgg ggagcggcga
     8101 ggaattgctc agcaagaatt atcatttgga gaacgaagtc gccaggctca agaaaggctc
     8161 ggggtcgggg gaggaattgt tgagtaaaaa ctaccacttg gaaaatgaag tcgccaggct
     8221 caaaaaaggg agtgggagcg gcgaagagtt attgagcaaa aattaccact tggagaacga
     8281 agtggcaagg ctcaagaaag ggagcggcag cggggaggag ctcttatcga agaactacca
     8341 cttagagaat gaagtcgccc gcttgaagaa aggctcgggg agcggggaag agctcttgag
     8401 caagaactac cacttggaaa atgaggtggc gcgcttgaag aaagggagcg ggagcgggga
     8461 agagttacta tctaagaatt atcatctcga gaacgaggtg gctcgactaa agaagggctc
     8521 cggcagtggg gaggaactcc tgtcgaagaa ctatcatctt gaaaatgagg ttgcaagact
     8581 taaaaagggg tccggatcag gtgaggaact actcagtaag aattaccacc tggaaaacga
     8641 agttgcacgt ttgaagaaag gatcaggatc aggcgaagaa ctgctctcaa aagattatca
     8701 tttggaaaat gaggttgcac gtttaaaaaa gggaagtggc agtggtgagg aacttctgtc
     8761 gaaaaattat catctcgaga atgaagtagc ccgacttaaa aagggttctc ccaagaagaa
     8821 gcgcaaggtg taagcggccg ctctggctct cgagtagggt aatctagata atcaacctct
     8881 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     8941 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     9001 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     9061 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     9121 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     9181 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     9241 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     9301 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     9361 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     9421 tcagacgagt cggatctccc tttgggccgc ctccccgcct gagatccttt aagaccaatg
     9481 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     9541 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     9601 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     9661 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     9721 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     9781 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggccc
     9841 gggttaatta aggaaagggc tagatcattc ttgaagacga aagggcctcg tgatacgcct
     9901 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     9961 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
    10021 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
    10081 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
    10141 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
    10201 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
    10261 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
    10321 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
    10381 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
    10441 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
    10501 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
    10561 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
    10621 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
    10681 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
    10741 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
    10801 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
    10861 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
    10921 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
    10981 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
    11041 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
    11101 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
    11161 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
    11221 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
    11281 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
    11341 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
    11401 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
    11461 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
    11521 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
    11581 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
    11641 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
    11701 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
    11761 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
    11821 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
    11881 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gcaagctcat
    11941 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
    12001 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct ccccgtggca
    12061 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
    12121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
    12181 tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga atttcacaaa
    12241 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
    12301 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
    12361 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
    12421 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
