LOCUS       sequence-107419-       11290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Sp dCas9-GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11290)
  AUTHORS   Ma H, Naseri A, Reyes-Gutierrez P, Wolfe SA, Zhang S, Pederson T
  TITLE     Multicolor CRISPR labeling of chromosomal loci in human cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Mar 10;112(10):3002-7. doi: 
            10.1073/pnas.1420024112. Epub 2015 Feb 23.
  PUBMED    25713381
REFERENCE   2  (bases 1 to 11290)
  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: 
            "10.1073/pnas.1420024112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-03-10- 10"; volume: "112"; issue: "10"; pages: "3002-7"
COMMENT     Sequence Label: pHAGE-SSFV-dCas9-GFP
FEATURES             Location/Qualifiers
     source          1..11290
                     /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"
     promoter        2277..2678
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      2712..2721
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2724..2744
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2763..2786,2787..2789,2790..5273,5274..5279,5280..5345,
                     5346..5348,5349..6863)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /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             join(2763..2786,2787..2789,2790..5276,5277..5279,
                     5280..6863)
                     /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             2763..6863
                     /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             6882..6902
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6915..7628
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    7680..8268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7733..7753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8151..8162)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8343..8576
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8695..8713)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8781..8885
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8886..8954,8955..9746)
                     /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(9104..9123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9917..10505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10406..10425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10659..10676
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10916..10937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10952..10957,10958..10975,10976..10982)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10990..11006
                     /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     10995..11017
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11014..11030
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11014..11030
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11080..11099
                     /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 gctagctgat gcgggccctg aaagacccca cctgtaggtt tggcaaccta gctgcagtaa
     2281 cgccattttg caaggcatgg aaaaatacca aaccaagaat agagaagttc agatcaaggg
     2341 cgggtacatg aaaatagcta acgttgggcc aaacaggata tctgcggtga gcagtttcgg
     2401 ccccggcccg gggccaagaa cagatggtca ccgcagtttc ggccccggcc cgaggccaag
     2461 aacagatggt ccccagatat ggcccaaccc tcagcagttt cttaagaccc atcagatgtt
     2521 tccaggctcc cccaaggacc tgaaatgacc ctgcgcctta tttgaattaa ccaatcagcc
     2581 tgcttctcgc ttctgttcgc gcgcttctgc ttcccgagct ctataaaaga gctcacaacc
     2641 cctcactcgg cgcgccagtc ctccgattga ctgagtcgcc caccggttaa ggtgaacgcg
     2701 tgcaggctgg cgccaccatg gcccctaaaa agaaacgcaa ggtcgaattc ggaagcggaa
     2761 gcgacaagaa gtactccatt gggctcgcta tcggtaccaa cagcgtcggc tgggccgtca
     2821 ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc
     2881 acagcataaa gaagaacctc attggagccc tcctgttcga ctccggggag acggccgaag
     2941 ccacgcggct caaaagaaca gcacggcgca gatatacccg cagaaagaat cggatctgct
     3001 acctgcagga gatctttagt aatgagatgg ctaaggtgga tgactctttc ttccataggc
     3061 tggaggagtc ctttttggtg gaggaggata aaaagcacga gcgccaccca atctttggca
     3121 atatcgtgga cgaggtggcg taccatgaaa agtacccaac catatatcat ctgaggaaga
     3181 agctggtaga cagtactgat aaggctgact tgcggttgat ctatctcgcg ctggcgcaca
     3241 tgatcaaatt tcggggacac ttcctcatcg agggggacct gaacccagac aacagcgatg
     3301 tcgacaaact ctttatccaa ctggttcaga cttacaatca gcttttcgag gagaacccga
     3361 tcaacgcatc cggcgttgac gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc
     3421 ggctcgaaaa cctcatcgca cagctccctg gggagaagaa gaacggcctg tttggtaatc
     3481 ttatcgccct gtcactcggg ctgaccccca actttaaatc taacttcgac ctggccgaag
     3541 atgccaagct gcaactgagc aaagacacct acgatgatga tctcgacaat ctgctggccc
     3601 agatcggcga ccagtacgca gacctttttt tggcggcaaa gaacctgtca gacgccattc
     3661 tgctgagtga tattctgcga gtgaacacgg agatcaccaa agctccgctg agcgctagta
     3721 tgatcaagcg ctatgatgag caccaccaag acttgacttt gctgaaggcc cttgtcagac
     3781 agcaactgcc tgagaagtac aaggaaattt tcttcgatca gtctaaaaat ggctacgccg
     3841 gatacattga cggcggagca agccaggagg aattttacaa atttattaag cccatcttgg
     3901 aaaaaatgga cggcaccgag gagctgctgg taaagctgaa cagagaagat ctgttgcgca
     3961 aacagcgcac tttcgacaat ggaagcatcc cccaccagat tcacctgggc gaactgcacg
     4021 ctatcctcag gcggcaagag gatttctacc cctttttgaa agataacagg gaaaagattg
     4081 agaaaatcct cacatttcgg ataccctact atgtaggccc cctcgctcgg ggaaattcca
     4141 gattcgcgtg gatgactcgc aaatcagaag agaccatcac tccctggaac ttcgaggaag
     4201 tcgtggataa gggggcctct gcccagtcct tcatcgaaag gatgactaac tttgataaaa
     4261 atctgcctaa cgaaaaggtg cttcctaaac actctctgct gtacgagtac ttcacagttt
     4321 ataacgagct caccaaggtc aaatacgtca cagaagggat gagaaagcca gcattcctgt
     4381 ctggagagca gaagaaagct atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg
     4441 tgaaacagct caaagaagac tatttcaaaa agattgaatg tttcgactct gttgaaatca
     4501 gcggagtgga ggatcgcttc aacgcatccc tgggaacgta tcacgatctc ctgaaaatca
     4561 ttaaagacaa ggacttcctg gacaatgagg agaacgagga cattcttgag gacattgtcc
     4621 tcacccttac gttgtttgaa gatagggaga tgattgaaga acgcttgaaa acttacgctc
     4681 atctcttcga cgacaaagtc atgaaacagc tcaagagacg ccgatataca ggatgggggc
     4741 ggctgtcaag aaaactgatc aatggcatcc gagacaagca gagtggaaag acaatcctgg
     4801 attttcttaa gtccgatgga tttgccaacc ggaacttcat gcagttgatc catgatgact
     4861 ctctcacctt taaggaggac atccagaaag cacaagtttc tggccagggg gacagtcttc
     4921 acgagcacat cgctaatctt gcaggtagcc cagctatcaa aaagggaata ctgcagaccg
     4981 ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca taagcccgag aatatcgtta
     5041 tcgagatggc ccgagagaac caaactaccc agaagggaca gaagaacagt agggaaagga
     5101 tgaagaggat tgaagagggt ataaaagaac tggggtccca aatccttaag gaacacccag
     5161 ttgaaaacac ccagcttcag aatgagaagc tctacctgta ctacctgcag aacggcaggg
     5221 acatgtacgt ggatcaggaa ctggacatca accggttgtc cgactacgac gtggatgcta
     5281 tcgtgcccca aagctttctc aaagatgatt ctattgataa taaagtgttg acaagatccg
     5341 ataaaaatag agggaagagt gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa
     5401 attattggcg gcagctgctg aacgccaaac tgatcacaca acggaagttc gataatctga
     5461 ctaaggctga acgaggtggc ctgtctgagt tggataaagc cggcttcatc aaaaggcagc
     5521 ttgttgagac acgccagatc accaagcacg tggcccaaat tctcgattca cgcatgaaca
     5581 ccaagtacga tgaaaatgac aaactgattc gagaggtgaa agttattact ctgaagtcta
     5641 agctggtctc agatttcaga aaggactttc agttttataa ggtgagagag atcaacaatt
     5701 accaccatgc gcatgatgcc tacctgaatg cagtggtagg cactgcactt atcaaaaaat
     5761 atcccaagct ggaatctgaa tttgtttacg gagactataa agtgtacgat gttaggaaaa
     5821 tgatcgcaaa gtctgagcag gaaataggca aggccaccgc taagtacttc ttttacagca
     5881 atattatgaa ttttttcaag accgagatta cactggccaa tggagagatt cggaagcgac
     5941 cacttatcga aacaaacgga gaaacaggag aaatcgtgtg ggacaagggt agggatttcg
     6001 cgacagtccg caaggtcctg tccatgccgc aggtgaacat cgttaaaaag accgaagtac
     6061 agaccggagg cttctccaag gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg
     6121 cacgcaaaaa agattgggac cccaagaaat acggcggatt cgattctcct acagtcgctt
     6181 acagtgtact ggttgtggcc aaagtggaga aagggaagtc taaaaaactc aaaagcgtca
     6241 aggaactgct gggcatcaca atcatggagc gatccagctt cgagaaaaac cccatcgact
     6301 ttctcgaagc gaaaggatat aaagaggtca aaaaagacct catcattaag ctgcccaagt
     6361 actctctctt tgagcttgaa aacggccgga aacgaatgct cgctagtgcg ggcgagctgc
     6421 agaaaggtaa cgagctggca ctgccctcta aatacgttaa tttcttgtat ctggccagcc
     6481 actatgaaaa gctcaaaggg tctcccgaag ataatgagca gaagcagctg ttcgtggaac
     6541 aacacaaaca ctaccttgat gagatcatcg agcaaataag cgagttctcc aaaagagtga
     6601 tcctcgccga cgctaacctc gataaggtgc tttctgctta caataagcac agggataagc
     6661 ccatcaggga gcaggcagaa aacattatcc acttgtttac tctgaccaac ttgggcgcgc
     6721 ctgcagcctt caagtacttc gacaccacca tagacagaaa gcggtacacc tctacaaagg
     6781 aggtcctgga cgccacactg attcatcagt caattacggg gctctatgaa acaagaatcg
     6841 acctctctca gctcggtgga gacggctcta ctagtggctc tcccaagaag aagaggaagg
     6901 taggatccgg aagtatgcgt aaaggcgaag agctgttcac tggtgtcgtc cctattctgg
     6961 tggaactgga tggtgatgtc aacggtcata agttttccgt gcgtggcgag ggtgaaggtg
     7021 acgcaactaa tggtaaactg acgctgaagt tcatctgtac tactggtaaa ctgccggtac
     7081 cttggccgac tctggtaacg acgctgactt atggtgttca gtgctttgct cgttatccgg
     7141 accatatgaa gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac
     7201 gcacgatttc ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag
     7261 gcgataccct ggtaaaccgc attgagctga aaggcattga ctttaaagaa gacggcaata
     7321 tcctgggcca taagctggaa tacaatttta acagccacaa tgtttacatc accgccgata
     7381 aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag gatggcagcg
     7441 tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc
     7501 cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg
     7561 atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac
     7621 tgtacaaatc tggcggccgc tctggcatcg atgggagcct cgagtagggt aatctagata
     7681 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     7741 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     7801 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     7861 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     7921 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     7981 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     8041 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     8101 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     8161 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     8221 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct gagatccttt
     8281 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     8341 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     8401 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     8461 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     8521 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     8581 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     8641 tgagaggccc gggttaatta aggaaagggc tagatcattc ttgaagacga aagggcctcg
     8701 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     8761 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     8821 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     8881 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     8941 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     9001 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     9061 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     9121 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     9181 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     9241 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     9301 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     9361 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     9421 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     9481 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     9541 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     9601 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     9661 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     9721 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     9781 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     9841 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     9901 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     9961 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
    10021 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
    10081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
    10141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
    10201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
    10261 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
    10321 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
    10381 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
    10441 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
    10501 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
    10561 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
    10621 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
    10681 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
    10741 gcaagctcat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
    10801 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
    10861 ccccgtggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
    10921 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
    10981 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga
    11041 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
    11101 atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac
    11161 ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct
    11221 gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa
    11281 acttagtagt
//
