LOCUS       sequence-308932-       10761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9 fused to 10 copies of the P3 coiled coil in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10761)
  AUTHORS   Lebar T, Lainscek D, Merljak E, Aupic J, Jerala R
  TITLE     A tunable orthogonal coiled-coil interaction toolbox for engineering
            mammalian cells.
  JOURNAL   Nat Chem Biol. 2020 Jan 6. pii: 10.1038/s41589-019-0443-y. doi: 
            10.1038/s41589-019-0443-y.
  PUBMED    31907374
REFERENCE   2  (bases 1 to 10761)
  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"; journalName: "Nat Chem 
            Biol. 2020 Jan 6. pii: 10.1038/s41589-019-0443-y. doi: 
            10.1038/s41589-019-0443-y."
COMMENT     Sequence Label: pcDNA3-His:dCas9:nls:10P3
FEATURES             Location/Qualifiers
     source          1..10761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             953..970
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(971..994,995..997,998..3484,3485..3487,3488..5068)
                     /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
                     ATLIHQSITGLYETRIDLSQLGG"
     CDS             971..5068
                     /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
                     ATLIHQSITGLYETRIDLSQLGG"
     CDS             5075..5095
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(6314..6332)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6315..6332)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6352..6369)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6358..6582
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6628..7056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6715..6734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6925..6946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7065..7085)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7070..7399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7250..7385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7312..7331
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7466..8260
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7520..7539)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8130..8149
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8434..8555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8471..8490)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8525..8544
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8604..8620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8604..8620)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8617..8639)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8628..8644
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8652..8658,8659..8676,8677..8682))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8697..8718
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8835..8852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9006..9594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9086..9105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9765..10556,10557..10625))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10388..10407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10626..10730)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgccacca tgcaccacca
      961 ccaccaccac gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg
     1021 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     1081 cgaccggcac agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggagaaac
     1141 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     1201 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     1261 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     1321 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     1381 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     1441 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     1501 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     1561 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     1621 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     1681 cggcaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     1741 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     1801 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     1861 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     1921 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     1981 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     2041 ctacgccggc tacatcgatg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     2101 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     2161 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     2221 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     2281 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     2341 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     2401 cgaggaagtg gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt
     2461 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     2521 caccgtgtac aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     2581 cttcctgagc ggcgagcaga aaaaagccat cgtggacctg ctgttcaaga ccaaccggaa
     2641 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     2701 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     2761 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     2821 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     2881 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     2941 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     3001 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     3061 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     3121 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     3181 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     3241 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     3301 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     3361 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     3421 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     3481 ggacgctatc gtgcctcaga gctttctgaa ggacgactcc atcgataaca aagtgctgac
     3541 tcggagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     3601 gatgaagaac tactggcgcc agctgctgaa tgccaagctg attacccaga ggaagttcga
     3661 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     3721 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     3781 gatgaacact aagtacgacg agaacgacaa actgatccgg gaagtgaaag tgatcaccct
     3841 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     3901 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     3961 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     4021 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     4081 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     4141 gaagcggcct ctgatcgaga caaacggcga aacaggcgag atcgtgtggg ataagggccg
     4201 ggactttgcc accgtgcgga aagtgctgtc tatgccccaa gtgaatatcg tgaaaaagac
     4261 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgacaa
     4321 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     4381 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     4441 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     4501 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     4561 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     4621 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     4681 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     4741 tgtggaacag cacaaacact acctggacga gatcatcgag cagatcagcg agttctccaa
     4801 gagagtgatc ctggccgacg ctaatctgga caaggtgctg agcgcctaca acaagcacag
     4861 agacaagcct atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     4921 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     4981 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     5041 acggatcgac ctgtctcagc tgggaggctc tgatccaaaa aagaagagaa aggtatcccc
     5101 ggaagatgag atccagcaac tggaagaaga aatcgctcag ctggaacaga aaaacgcagc
     5161 gctgaaagag aaaaaccagg cgctgaaata cggtcttaag agcggccccg gcagcggctc
     5221 cccggaagat gagatccagc aactggaaga agaaatcgct cagctggaac agaaaaacgc
     5281 agcgctgaaa gagaaaaacc aggcgctgaa atacggtacc ggtagcggcc ccggcagcgg
     5341 ctccccggaa gatgagatcc agcaactgga agaagaaatc gctcagctgg aacagaaaaa
     5401 cgcagcgctg aaagagaaaa accaggcgct gaaatacggt gctagcagcg gccccggcag
     5461 cggctccccg gaagatgaga tccagcaact ggaagaagaa atcgctcagc tggaacagaa
     5521 aaacgcagcg ctgaaagaga aaaaccaggc gctgaaatac ggtctcgaga gcggccccgg
     5581 cagcggctcc ccggaagatg agatccagca actggaagaa gaaatcgctc agctggaaca
     5641 gaaaaacgca gcgctgaaag agaaaaacca ggcgctgaaa tacggtagcg gccccggcag
     5701 cggctccccg gaagatgaga tccagcaact ggaagaagaa atcgctcagc tggaacagaa
     5761 aaacgcagcg ctgaaagaga aaaaccaggc gctgaaatac ggtctcgaga gcggccccgg
     5821 cagcggctcc ccggaagatg agatccagca actggaagaa gaaatcgctc agctggaaca
     5881 gaaaaacgca gcgctgaaag agaaaaacca ggcgctgaaa tacggtgcta gcagcggccc
     5941 cggcagcggc tccccggaag atgagatcca gcaactggaa gaagaaatcg ctcagctgga
     6001 acagaaaaac gcagcgctga aagagaaaaa ccaggcgctg aaatacggta ccggtagcgg
     6061 ccccggcagc ggctccccgg aagatgagat ccagcaactg gaagaagaaa tcgctcagct
     6121 ggaacagaaa aacgcagcgc tgaaagagaa aaaccaggcg ctgaaatacg gtcttaagag
     6181 cggccccggc agcggctccc cggaagatga gatccagcaa ctggaagaag aaatcgctca
     6241 gctggaacag aaaaacgcag cgctgaaaga gaaaaaccag gcgctgaaat acggttaatc
     6301 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     6361 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     6421 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     6481 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     6541 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     6601 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     6661 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     6721 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     6781 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     6841 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccttttga
     6901 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     6961 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     7021 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     7081 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     7141 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     7201 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     7261 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     7321 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     7381 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     7441 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     7501 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     7561 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     7621 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     7681 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     7741 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     7801 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     7861 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     7921 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     7981 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     8041 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     8101 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     8161 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     8221 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     8281 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     8341 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     8401 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     8461 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     8521 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     8581 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     8641 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     8701 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     8761 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     8821 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     8881 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     8941 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     9001 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     9061 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     9121 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     9181 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     9241 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     9301 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     9361 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     9421 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     9481 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     9541 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     9601 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     9661 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     9721 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     9781 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     9841 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     9901 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     9961 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
    10021 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
    10081 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
    10141 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
    10201 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
    10261 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
    10321 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    10381 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    10441 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    10501 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    10561 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    10621 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
    10681 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
    10741 cgaaaagtgc cacctgacgt c
//
