LOCUS       sequence-264371-       10050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpyCas9 fused to attenuated AcrIIA4 (D14A/G38A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10050)
  AUTHORS   Aschenbrenner S, Kallenberger SM, Hoffmann MD, Huck A, Eils R, 
            Niopek D
  TITLE     Coupling Cas9 to artificial inhibitory domains enhances CRISPR-Cas9 
            target specificity.
  JOURNAL   Sci Adv. 2020 Feb 5;6(6):eaay0187. doi: 10.1126/sciadv.aay0187. 
            eCollection 2020 Feb.
  PUBMED    32076642
REFERENCE   2  (bases 1 to 10050)
  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: "Sci Adv."; 
            date: "2020-02-5"; volume: "6(6):eaay0187. doi"; pages: " 
            10.1126/sciadv.aay0187. eCollection 2020 Feb"
COMMENT     Sequence Label: Cas-Acr (D14A/G38A)
FEATURES             Location/Qualifiers
     source          1..10050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..83)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        255..634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        635..838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     789..809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     883..902
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        883..901
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      951..960
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             960..1025
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1032..1052
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1077..5177
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1077..5177
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5178..5225
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5661..5678)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5667..5891
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5937..6365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6024..6043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6234..6255
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6374..6394)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6379..6708
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6559..6694
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6621..6640
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6775..7569
                     /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(6829..6848)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7439..7458
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7743..7864
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7780..7799)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7834..7853
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7913..7929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7913..7929)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7926..7948)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7937..7953
                     /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(7961..7967,7968..7985,7986..7991))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8006..8027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8144..8161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8315..8903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8395..8414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9074..9865,9866..9934))
                     /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     9697..9716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9935..10039)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
       61 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
      121 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
      181 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
      241 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      301 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      361 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      421 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      481 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      541 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      601 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      661 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      721 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      781 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
      841 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
      901 ggagacccaa gctggctagc gtttaaacgg gccctctaga ctcgagcggt gccaccatgg
      961 actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg
     1021 ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca
     1081 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
     1141 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
     1201 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
     1261 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
     1321 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
     1381 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
     1441 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
     1501 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
     1561 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
     1621 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     1681 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     1741 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
     1801 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     1861 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     1921 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     1981 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     2041 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     2101 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     2161 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     2221 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     2281 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     2341 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     2401 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     2461 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     2521 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     2581 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     2641 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     2701 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     2761 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     2821 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     2881 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     2941 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     3001 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     3061 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     3121 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     3181 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     3241 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     3301 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     3361 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     3421 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     3481 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     3541 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     3601 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     3661 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     3721 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     3781 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     3841 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     3901 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     3961 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     4021 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     4081 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     4141 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     4201 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     4261 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     4321 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     4381 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     4441 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     4501 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     4561 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     4621 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     4681 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     4741 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     4801 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     4861 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     4921 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     4981 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     5041 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     5101 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     5161 ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
     5221 aaaaggaatt cgggggatca gggggagggt ctggtggtgg atctggtgga ggatctggtg
     5281 ggggttcagg gggcggttcc ggaggaggct caggaggcgg ctccggcggt ggctcaggag
     5341 gtggatccgg tatgaacatt aacgacctga ttcgggaaat aaagaacaag gcctacacgg
     5401 tcaagctgtc aggcaccgac tccaactcta taacacagct cattataagg gttaataatg
     5461 atgccaacga atatgtcatc tctgaatccg agaatgaatc aatcgtggag aaattcatca
     5521 gtgcatttaa aaacggctgg aaccaggagt acgaagatga ggaagagttc tacaatgata
     5581 tgcaaactat cacactgaaa tctgagctga actagctgaa gttcatctgc accaaagctt
     5641 aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     5701 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     5761 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     5821 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     5881 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     5941 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     6001 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     6061 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     6121 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     6181 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6241 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6301 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6361 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6481 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6541 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6601 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     6661 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     6721 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     6781 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     6841 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     6901 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     6961 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     7021 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     7081 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     7141 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     7201 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     7261 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     7321 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     7381 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     7441 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     7501 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     7561 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     7621 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     7681 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     7741 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7801 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7861 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     7921 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7981 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     8041 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     8101 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     8161 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     8221 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     8281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     8341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     8401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     8461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     8521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     8581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     8641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     8761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8821 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8881 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8941 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     9001 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     9061 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     9121 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     9181 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     9241 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     9301 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     9361 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     9421 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     9481 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     9541 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     9601 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     9661 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9721 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9781 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9841 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9901 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9961 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
    10021 aacaaatagg ggttccgcgc acatttcccc
//
