LOCUS       sequence-107420-       11552 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 11552)
  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 11552)
  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-TO-dCas9-GFP
FEATURES             Location/Qualifiers
     source          1..11552
                     /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        2299..2602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2603..2806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2757..2777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2808..2826
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2857..2881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2974..2983
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2986..3006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3025..3048,3049..3051,3052..5535,5536..5541,5542..5607,
                     5608..5610,5611..7125)
                     /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(3025..3048,3049..3051,3052..5538,5539..5541,
                     5542..7125)
                     /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             3025..7125
                     /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             7144..7164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7177..7890
                     /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    7942..8530
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7995..8015)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8413..8424)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8605..8838
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8957..8975)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9043..9147
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9148..9216,9217..10008)
                     /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(9366..9385)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10179..10767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10668..10687
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10921..10938
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11178..11199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11214..11219,11220..11237,11238..11244)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11252..11268
                     /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     11257..11279
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11276..11292
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11276..11292
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11342..11361
                     /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 gcgggcccta gttattaata gtaatcaatt acggggtcat tagttcatag
     2281 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2341 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2401 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2461 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2521 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2581 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2641 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2701 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     2761 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga
     2821 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat
     2881 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     2941 cctaccggtt aaggtgaacg cgtgcaggct ggcgccacca tggcccctaa aaagaaacgc
     3001 aaggtcgaat tcggaagcgg aagcgacaag aagtactcca ttgggctcgc tatcggtacc
     3061 aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa
     3121 gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggagc cctcctgttc
     3181 gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc
     3241 cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg
     3301 gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac
     3361 gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca
     3421 accatatatc atctgaggaa gaagctggta gacagtactg ataaggctga cttgcggttg
     3481 atctatctcg cgctggcgca catgatcaaa tttcggggac acttcctcat cgagggggac
     3541 ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat
     3601 cagcttttcg aggagaaccc gatcaacgca tccggcgttg acgccaaagc aatcctgagc
     3661 gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag
     3721 aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa
     3781 tctaacttcg acctggccga agatgccaag ctgcaactga gcaaagacac ctacgatgat
     3841 gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca
     3901 aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc
     3961 aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact
     4021 ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat
     4081 cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac
     4141 aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctg
     4201 aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag
     4261 attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg
     4321 aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc
     4381 cccctcgctc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc
     4441 actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa
     4501 aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg
     4561 ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg
     4621 atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc
     4681 aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa
     4741 tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg
     4801 tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag
     4861 gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa
     4921 gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagaga
     4981 cgccgatata caggatgggg gcggctgtca agaaaactga tcaatggcat ccgagacaag
     5041 cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc
     5101 atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt
     5161 tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc
     5221 aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg
     5281 cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga
     5341 cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc
     5401 caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg
     5461 tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caaccggttg
     5521 tccgactacg acgtggatgc tatcgtgccc caaagctttc tcaaagatga ttctattgat
     5581 aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa
     5641 gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca
     5701 caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa
     5761 gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa
     5821 attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg
     5881 aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat
     5941 aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta
     6001 ggcactgcac ttatcaaaaa atatcccaag ctggaatctg aatttgttta cggagactat
     6061 aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc
     6121 gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc
     6181 aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg
     6241 tgggacaagg gtagggattt cgcgacagtc cgcaaggtcc tgtccatgcc gcaggtgaac
     6301 atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa
     6361 aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga
     6421 ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag
     6481 tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatccagc
     6541 ttcgagaaaa accccatcga ctttctcgaa gcgaaaggat ataaagaggt caaaaaagac
     6601 ctcatcatta agctgcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg
     6661 ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt
     6721 aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag
     6781 cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata
     6841 agcgagttct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct
     6901 tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt
     6961 actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga
     7021 aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg
     7081 gggctctatg aaacaagaat cgacctctct cagctcggtg gagacggctc tactagtggc
     7141 tctcccaaga agaagaggaa ggtaggatcc ggaagtatgc gtaaaggcga agagctgttc
     7201 actggtgtcg tccctattct ggtggaactg gatggtgatg tcaacggtca taagttttcc
     7261 gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt
     7321 actactggta aactgccggt accttggccg actctggtaa cgacgctgac ttatggtgtt
     7381 cagtgctttg ctcgttatcc ggaccatatg aagcagcatg acttcttcaa gtccgccatg
     7441 ccggaaggct atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg
     7501 cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt
     7561 gactttaaag aagacggcaa tatcctgggc cataagctgg aatacaattt taacagccac
     7621 aatgtttaca tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc
     7681 cacaacgtgg aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc
     7741 ggtgatggtc ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct
     7801 aaagatccga acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc
     7861 atcacgcatg gtatggatga actgtacaaa tctggcggcc gctctggcat cgatgggagc
     7921 ctcgagtagg gtaatctaga taatcaacct ctggattaca aaatttgtga aagattgact
     7981 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     8041 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     8101 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     8161 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     8221 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     8281 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     8341 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     8401 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     8461 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     8521 gcctccccgc ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     8581 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     8641 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     8701 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     8761 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     8821 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     8881 caaagaaatg aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat
     8941 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     9001 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     9061 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     9121 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     9181 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     9241 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     9301 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     9361 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     9421 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     9481 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     9541 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     9601 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     9661 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     9721 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     9781 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     9841 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     9901 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     9961 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
    10021 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
    10081 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
    10141 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
    10201 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
    10261 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
    10321 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
    10381 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
    10441 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
    10501 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
    10561 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
    10621 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
    10681 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
    10741 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
    10801 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
    10861 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
    10921 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
    10981 cgttggccga ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag
    11041 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
    11101 gcctaggctt ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc
    11161 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
    11221 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
    11281 aacagctatg acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt
    11341 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
    11401 gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg
    11461 tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt
    11521 atttgttaaa tatgtactac aaacttagta gt
//
