LOCUS       sequence-107303-       11099 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 11099)
  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 11099)
  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-EFS-dCas9-GFP
FEATURES             Location/Qualifiers
     source          1..11099
                     /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        2265..2476
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2521..2530
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2533..2553
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2572..2595,2596..2598,2599..5082,5083..5088,5089..5154,
                     5155..5157,5158..6672)
                     /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(2572..2595,2596..2598,2599..5085,5086..5088,
                     5089..6672)
                     /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             2572..6672
                     /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             6691..6711
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6724..7437
                     /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    7489..8077
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7542..7562)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7960..7971)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8152..8385
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8504..8522)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8590..8694
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8695..8763,8764..9555)
                     /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(8913..8932)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9726..10314
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10215..10234
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10468..10485
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10725..10746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10761..10766,10767..10784,10785..10791)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10799..10815
                     /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     10804..10826
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10823..10839
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10823..10839
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10889..10908
                     /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 gcgggccctt cgagtggctc cggtgcccgt cagtgggcag agcgcacatc
     2281 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     2341 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     2401 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     2461 tgccgccaga acacaggtgt cgtgacgcgg tccggttaag gtgaacgcgt gcaggctggc
     2521 gccaccatgg cccctaaaaa gaaacgcaag gtcgaattcg gaagcggaag cgacaagaag
     2581 tactccattg ggctcgctat cggtaccaac agcgtcggct gggccgtcat tacggacgag
     2641 tacaaggtgc cgagcaaaaa attcaaagtt ctgggcaata ccgatcgcca cagcataaag
     2701 aagaacctca ttggagccct cctgttcgac tccggggaga cggccgaagc cacgcggctc
     2761 aaaagaacag cacggcgcag atatacccgc agaaagaatc ggatctgcta cctgcaggag
     2821 atctttagta atgagatggc taaggtggat gactctttct tccataggct ggaggagtcc
     2881 tttttggtgg aggaggataa aaagcacgag cgccacccaa tctttggcaa tatcgtggac
     2941 gaggtggcgt accatgaaaa gtacccaacc atatatcatc tgaggaagaa gctggtagac
     3001 agtactgata aggctgactt gcggttgatc tatctcgcgc tggcgcacat gatcaaattt
     3061 cggggacact tcctcatcga gggggacctg aacccagaca acagcgatgt cgacaaactc
     3121 tttatccaac tggttcagac ttacaatcag cttttcgagg agaacccgat caacgcatcc
     3181 ggcgttgacg ccaaagcaat cctgagcgct aggctgtcca aatcccggcg gctcgaaaac
     3241 ctcatcgcac agctccctgg ggagaagaag aacggcctgt ttggtaatct tatcgccctg
     3301 tcactcgggc tgacccccaa ctttaaatct aacttcgacc tggccgaaga tgccaagctg
     3361 caactgagca aagacaccta cgatgatgat ctcgacaatc tgctggccca gatcggcgac
     3421 cagtacgcag accttttttt ggcggcaaag aacctgtcag acgccattct gctgagtgat
     3481 attctgcgag tgaacacgga gatcaccaaa gctccgctga gcgctagtat gatcaagcgc
     3541 tatgatgagc accaccaaga cttgactttg ctgaaggccc ttgtcagaca gcaactgcct
     3601 gagaagtaca aggaaatttt cttcgatcag tctaaaaatg gctacgccgg atacattgac
     3661 ggcggagcaa gccaggagga attttacaaa tttattaagc ccatcttgga aaaaatggac
     3721 ggcaccgagg agctgctggt aaagctgaac agagaagatc tgttgcgcaa acagcgcact
     3781 ttcgacaatg gaagcatccc ccaccagatt cacctgggcg aactgcacgc tatcctcagg
     3841 cggcaagagg atttctaccc ctttttgaaa gataacaggg aaaagattga gaaaatcctc
     3901 acatttcgga taccctacta tgtaggcccc ctcgctcggg gaaattccag attcgcgtgg
     3961 atgactcgca aatcagaaga gaccatcact ccctggaact tcgaggaagt cgtggataag
     4021 ggggcctctg cccagtcctt catcgaaagg atgactaact ttgataaaaa tctgcctaac
     4081 gaaaaggtgc ttcctaaaca ctctctgctg tacgagtact tcacagttta taacgagctc
     4141 accaaggtca aatacgtcac agaagggatg agaaagccag cattcctgtc tggagagcag
     4201 aagaaagcta tcgtggacct cctcttcaag acgaaccgga aagttaccgt gaaacagctc
     4261 aaagaagact atttcaaaaa gattgaatgt ttcgactctg ttgaaatcag cggagtggag
     4321 gatcgcttca acgcatccct gggaacgtat cacgatctcc tgaaaatcat taaagacaag
     4381 gacttcctgg acaatgagga gaacgaggac attcttgagg acattgtcct cacccttacg
     4441 ttgtttgaag atagggagat gattgaagaa cgcttgaaaa cttacgctca tctcttcgac
     4501 gacaaagtca tgaaacagct caagagacgc cgatatacag gatgggggcg gctgtcaaga
     4561 aaactgatca atggcatccg agacaagcag agtggaaaga caatcctgga ttttcttaag
     4621 tccgatggat ttgccaaccg gaacttcatg cagttgatcc atgatgactc tctcaccttt
     4681 aaggaggaca tccagaaagc acaagtttct ggccaggggg acagtcttca cgagcacatc
     4741 gctaatcttg caggtagccc agctatcaaa aagggaatac tgcagaccgt taaggtcgtg
     4801 gatgaactcg tcaaagtaat gggaaggcat aagcccgaga atatcgttat cgagatggcc
     4861 cgagagaacc aaactaccca gaagggacag aagaacagta gggaaaggat gaagaggatt
     4921 gaagagggta taaaagaact ggggtcccaa atccttaagg aacacccagt tgaaaacacc
     4981 cagcttcaga atgagaagct ctacctgtac tacctgcaga acggcaggga catgtacgtg
     5041 gatcaggaac tggacatcaa ccggttgtcc gactacgacg tggatgctat cgtgccccaa
     5101 agctttctca aagatgattc tattgataat aaagtgttga caagatccga taaaaataga
     5161 gggaagagtg ataacgtccc ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg
     5221 cagctgctga acgccaaact gatcacacaa cggaagttcg ataatctgac taaggctgaa
     5281 cgaggtggcc tgtctgagtt ggataaagcc ggcttcatca aaaggcagct tgttgagaca
     5341 cgccagatca ccaagcacgt ggcccaaatt ctcgattcac gcatgaacac caagtacgat
     5401 gaaaatgaca aactgattcg agaggtgaaa gttattactc tgaagtctaa gctggtctca
     5461 gatttcagaa aggactttca gttttataag gtgagagaga tcaacaatta ccaccatgcg
     5521 catgatgcct acctgaatgc agtggtaggc actgcactta tcaaaaaata tcccaagctg
     5581 gaatctgaat ttgtttacgg agactataaa gtgtacgatg ttaggaaaat gatcgcaaag
     5641 tctgagcagg aaataggcaa ggccaccgct aagtacttct tttacagcaa tattatgaat
     5701 tttttcaaga ccgagattac actggccaat ggagagattc ggaagcgacc acttatcgaa
     5761 acaaacggag aaacaggaga aatcgtgtgg gacaagggta gggatttcgc gacagtccgc
     5821 aaggtcctgt ccatgccgca ggtgaacatc gttaaaaaga ccgaagtaca gaccggaggc
     5881 ttctccaagg aaagtatcct cccgaaaagg aacagcgaca agctgatcgc acgcaaaaaa
     5941 gattgggacc ccaagaaata cggcggattc gattctccta cagtcgctta cagtgtactg
     6001 gttgtggcca aagtggagaa agggaagtct aaaaaactca aaagcgtcaa ggaactgctg
     6061 ggcatcacaa tcatggagcg atccagcttc gagaaaaacc ccatcgactt tctcgaagcg
     6121 aaaggatata aagaggtcaa aaaagacctc atcattaagc tgcccaagta ctctctcttt
     6181 gagcttgaaa acggccggaa acgaatgctc gctagtgcgg gcgagctgca gaaaggtaac
     6241 gagctggcac tgccctctaa atacgttaat ttcttgtatc tggccagcca ctatgaaaag
     6301 ctcaaagggt ctcccgaaga taatgagcag aagcagctgt tcgtggaaca acacaaacac
     6361 taccttgatg agatcatcga gcaaataagc gagttctcca aaagagtgat cctcgccgac
     6421 gctaacctcg ataaggtgct ttctgcttac aataagcaca gggataagcc catcagggag
     6481 caggcagaaa acattatcca cttgtttact ctgaccaact tgggcgcgcc tgcagccttc
     6541 aagtacttcg acaccaccat agacagaaag cggtacacct ctacaaagga ggtcctggac
     6601 gccacactga ttcatcagtc aattacgggg ctctatgaaa caagaatcga cctctctcag
     6661 ctcggtggag acggctctac tagtggctct cccaagaaga agaggaaggt aggatccgga
     6721 agtatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat
     6781 ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat
     6841 ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggtacc ttggccgact
     6901 ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag
     6961 cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc
     7021 tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg cgataccctg
     7081 gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat cctgggccat
     7141 aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa acaaaaaaat
     7201 ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt gcagctggct
     7261 gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc agacaatcac
     7321 tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga tcatatggtt
     7381 ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact gtacaaatct
     7441 ggcggccgct ctggcatcga tgggagcctc gagtagggta atctagataa tcaacctctg
     7501 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     7561 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     7621 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     7681 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     7741 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     7801 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     7861 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     7921 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     7981 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     8041 cagacgagtc ggatctccct ttgggccgcc tccccgcctg agatccttta agaccaatga
     8101 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     8161 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     8221 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     8281 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     8341 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     8401 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcccg
     8461 ggttaattaa ggaaagggct agatcattct tgaagacgaa agggcctcgt gatacgccta
     8521 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     8581 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     8641 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     8701 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     8761 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     8821 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     8881 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     8941 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     9001 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     9061 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     9121 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     9181 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     9241 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     9301 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     9361 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     9421 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     9481 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     9541 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     9601 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     9661 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     9721 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     9781 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     9841 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     9901 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     9961 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
    10021 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
    10081 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
    10141 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
    10201 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
    10261 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
    10321 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
    10381 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
    10441 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
    10501 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag caagctcatg
    10561 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
    10621 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc cccgtggcac
    10681 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
    10741 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
    10801 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa tttcacaaat
    10861 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
    10921 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
    10981 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
    11041 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
