LOCUS       sequence-317143-        9639 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express nuclease dead Cas9 fused to KillerRed chromophore in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9639)
  AUTHORS   House NCM, Parasuram R, Layer JV, Price BD
  TITLE     Site-specific targeting of a light activated dCas9-KillerRed fusion 
            protein generates transient, localized regions of oxidative DNA 
            damage.
  JOURNAL   PLoS One. 2020 Dec 17;15(12):e0237759. doi: 
            10.1371/journal.pone.0237759. eCollection 2020.
  PUBMED    33332350
REFERENCE   2  (bases 1 to 9639)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2020-12-17"; volume: "15(12):e0237759. doi"; pages: " 
            10.1371/journal.pone.0237759. eCollection 2020"
COMMENT     Sequence Label: pJ609-dCas9-KR-puro
FEATURES             Location/Qualifiers
     source          1..9639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(671..697,698..700,701..3184,3185..3190,3191..3256,
                     3257..3259,3260..4774)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4787..4807
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      4817..4826
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4823..4825,4826..4831,4832..5542)
                     /codon_start=1
                     /product="red fluorescent protein KillerRed, a 
                     genetically-encoded photosensitizer (Bulina et al., 2006)"
                     /label=KillerRed
                     /note="mammalian codon-optimized"
                     /translation="MGSEGGPALFQSDMTFKIFIDGEVNGQKFTIVADGSSKFPHGDFN
                     VHAVCETGKLPMSWKPICHLIQYGEPFFARYPDGISHFAQECFPEGLSIDRTVRFENDG
                     TMTSHHTYELDDTCVVSRITVNCDGFQPDGPIMRDQLVDILPNETHMFPHGPNAVRQLA
                     FIGFTTADGGLMMGHFDSKMTFNGSRAIEIPGPHFVTIITKQMRDTSDKRDHVCQREVA
                     YAHSVPRITSAIGSDED"
     primer_bind     complement(5557..5574)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5563..5674
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5792..5812)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5797..6126
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5977..6112
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6039..6058
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6174..6221
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6240..6839
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6240..6259)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6736..6756
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6969..7090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7006..7025)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7060..7079
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7143..7934,7935..8003))
                     /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     7766..7785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8004..8095)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8137..8156
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8162..8181
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      8267..8855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8756..8775
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9009..9026
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     9124..9141
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9125..9141
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9597..9613
                     /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)."
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggga agctggctag
      661 cgccgccacc atggacaaga agtactccat tgggctcgct atcggcacaa acagcgtcgg
      721 ctgggccgtc attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa
      781 taccgatcgc cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga
      841 gacggccgaa gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa
      901 tcggatctgc tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt
      961 cttccatagg ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc
     1021 aatctttggc aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca
     1081 tctgaggaag aagcttgtag acagtactga taaggctgac ttgcggttga tctatctcgc
     1141 gctggcgcat atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga
     1201 caacagcgat gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga
     1261 agagaacccg atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc
     1321 caaatcccgg cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct
     1381 gtttggtaat cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga
     1441 cctggccgaa gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa
     1501 tctgctggcc cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc
     1561 agacgccatt ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct
     1621 gagcgctagt atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc
     1681 ccttgtcaga cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa
     1741 tggctacgcc ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa
     1801 gcccatcttg gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga
     1861 tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg
     1921 cgaactgcac gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag
     1981 ggaaaagatt gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg
     2041 gggaaattcc agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa
     2101 cttcgaggaa gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa
     2161 ctttgataaa aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta
     2221 cttcacagtt tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc
     2281 agcattcctg tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg
     2341 gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc
     2401 tgttgaaatc agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct
     2461 cctgaaaatc attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga
     2521 ggacattgtc ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa
     2581 aacttacgct catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac
     2641 aggatggggg cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa
     2701 gacaatcctg gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat
     2761 ccatgatgac tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg
     2821 ggacagtctt cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat
     2881 actgcagacc gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga
     2941 gaatatcgtt atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag
     3001 tagggaaagg atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa
     3061 ggaacaccca gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca
     3121 gaacggcagg gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga
     3181 cgtggctgct atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt
     3241 gacaagatcc gataaagcta gagggaagag tgataacgtc ccctcagaag aagttgtcaa
     3301 gaaaatgaaa aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt
     3361 cgataatctg actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat
     3421 caaaaggcag cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc
     3481 acgcatgaac accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac
     3541 tctgaagtct aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga
     3601 gatcaacaat taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact
     3661 tatcaaaaaa tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga
     3721 tgttaggaaa atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt
     3781 cttttacagc aatattatga attttttcaa gaccgagatt acactggcca atggagagat
     3841 tcggaagcga ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg
     3901 tagggatttc gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa
     3961 gaccgaagta cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga
     4021 caagctgatc gcacgcaaaa aagattggga ccccaagaaa tacggcggat tcgattctcc
     4081 tacagtcgct tacagtgtac tggttgtggc caaagtggag aaagggaagt ctaaaaaact
     4141 caaaagcgtc aaggaactgc tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa
     4201 ccccatcgac tttctcgagg cgaaaggata taaagaggtc aaaaaagacc tcatcattaa
     4261 gcttcccaag tactctctct ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc
     4321 gggcgagctg cagaaaggta acgagctggc actgccctct aaatacgtta atttcttgta
     4381 tctggccagc cactatgaaa agctcaaagg gtctcccgaa gataatgagc agaagcagct
     4441 gttcgtggaa caacacaaac actaccttga tgagatcatc gagcaaataa gcgaattctc
     4501 caaaagagtg atcctcgccg acgctaacct cgataaggtg ctttctgctt acaataagca
     4561 cagggataag cccatcaggg agcaggcaga aaacattatc cacttgttta ctctgaccaa
     4621 cttgggcgcg cctgcagcct tcaagtactt cgacaccacc atagacagaa agcggtacac
     4681 ctctacaaag gaggtcctgg acgccacact gattcatcag tcaattacgg ggctctatga
     4741 aacaagaatc gacctctctc agctcggtgg agacagcagg gctgacccca agaagaagag
     4801 gaaggtgaaa ccggtcgcca ccatgggttc agagggcggc cccgccctgt tccagagcga
     4861 catgaccttc aaaatcttca tcgacggcga ggtgaacggc cagaagttca ccatcgtggc
     4921 cgacggcagc agcaagttcc cccacggcga cttcaacgtg cacgccgtgt gcgagaccgg
     4981 caagctgccc atgagctgga agcccatctg ccacctgatc cagtacggcg agcccttctt
     5041 cgcccgctac cccgacggca tcagccattt cgcccaggag tgcttccccg agggcctgag
     5101 catcgaccgc accgtgcgct tcgagaacga cggcaccatg accagccacc acacctacga
     5161 gctggacgac acctgcgtgg tgagccgcat caccgtgaac tgcgacggct tccagcccga
     5221 cggccccatc atgcgcgacc agctggtgga catcctgccc aacgagaccc acatgttccc
     5281 ccacggcccc aacgccgtgc gccagctggc cttcatcggc ttcaccaccg ccgacggcgg
     5341 cctgatgatg ggccacttcg acagcaagat gaccttcaac ggcagccgcg ccatcgagat
     5401 ccccggccca cacttcgtga ccatcatcac caagcagatg agggacacca gcgacaagcg
     5461 cgaccacgtg tgccagcgcg aggtggccta cgcccacagc gtgccccgca tcaccagcgc
     5521 catcggtagc gacgaggatt aagaattcaa aatcagcctc gactgtgcct tctagttgcc
     5581 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5641 ctgtcctttc ctaataaaat gaggaaattg catcacaaca ctcaacccta tctcggtcta
     5701 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5761 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     5821 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5881 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5941 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6001 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6061 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     6121 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca
     6181 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
     6241 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     6301 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     6361 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     6421 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     6481 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     6541 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     6601 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     6661 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     6721 agacatccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     6781 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     6841 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6901 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6961 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     7021 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     7081 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     7141 cattaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7201 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7261 cccagcgctg cgatgatacc gcgagaacca cgctcaccgg ctccggattt atcagcaata
     7321 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7381 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7441 aacgttgttg ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7501 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7561 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7621 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7681 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7741 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7801 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7861 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7921 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7981 acacggaaat gttgaatact catattcttc ctttttcaat attattgaag catttatcag
     8041 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8101 gtcagtgtta caaccaatta accaattctg aacattatcg cgagcccatt tatacctgaa
     8161 tatggctcat aacacccctt gctcatgacc aaaatccctt aacgtgagtt acgcgcgcgt
     8221 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     8281 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     8341 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     8401 taccaaatac tgttcttcta gtgtagccgt agttagccca ccacttcaag aactctgtag
     8461 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8521 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8581 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8641 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8701 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8761 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8821 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8881 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8941 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     9001 ccgagcgcag cgagtcagtg agcgaggaag cggaaggcga gagtagggaa ctgccaggca
     9061 tcaaactaag cagaaggccc ctgacggatg gcctttttgc gtttctacaa actctttctg
     9121 tgttgtaaaa cgacggccag tcttaagctc gggccccctg ggcggttctg ataacgagta
     9181 atcgttaatc cgcaaataac gtaaaaaccc gcttcggcgg gtttttttat ggggggagtt
     9241 tagggaaaga gcatttgtca gaatatttaa gggcgcctgt cactttgctt gatatatgag
     9301 aattatttaa ccttataaat gagaaaaaag caacgcactt taaataagat acgttgcttt
     9361 ttcgattgat gaacacctat aattaaacta ttcatctatt atttatgatt ttttgtatat
     9421 acaatatttc tagtttgtta aagagaatta agaaaataaa tctcgaaaat aataaaggga
     9481 aaatcagttt ttgatatcaa aattatacat gtcaacgata atacaaaata taatacaaac
     9541 tataagatgt tatcagtatt tattatcatt tagaataaat tttgtgtcgc ccttaattgt
     9601 gagcggataa caattacgag cttcatgcac agtggcgtt
//
