LOCUS       sequence-492667-       15807 bp    DNA     circular SYN 12-NOV-2025
DEFINITION  Expresses dCas9 under control of Hsp70 promoter and gRNAs targeting 
            Lucilia cuprina transformer gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15807)
  TITLE     bioRxiv -- Conditional expression of Cas9 and dCas9 in Lucilia 
            cuprina reveals dCas9-associated lethality
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 15807)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 12, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: dCas9-tra246
FEATURES             Location/Qualifiers
     source          1..15807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    265..364
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     protein_bind    499..517
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    520..538
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    541..559
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    562..580
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    583..601
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    604..622
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    625..643
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    646..664
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    667..685
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    688..706
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    816..834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    866..884
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1200..1247
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(1248..1271,1272..1274,1275..3830,3831..3833,
                     3834..5348)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5403..5423
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_RNA        complement(6449..6524)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(6623..6698)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(6797..6872)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    10639..10738
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     repeat_region   12289..12351
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(12408..12425)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12408..12424)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12417..12439)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(12633..12652)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12752..12774
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(12812..12830)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        12898..13002
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(13003..13071,13072..13863)
                     /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     complement(13221..13240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      14034..14622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14523..14542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     14776..14793
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    14910..14931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(14946..14951,14952..14969,14970..14976)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14984..15000
                     /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     14989..15011
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     15008..15024
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15008..15024
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   15085..15119
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 aacattcatg tgtgggtgat tacactcaca caaaaacaca cacattcaca caaacacaca
       61 cacttataaa gatgagaaca tttaacaaaa acaattataa aactgatggc aatggagcaa
      121 tgatgtgatg aagctgatga aagttttgaa tattcattca agagttaaaa acatgaacaa
      181 acatgcacaa aattagtcga gactgacgga cacaccgaag ccccggcggc aaccctcagc
      241 ggatgccccg gggcttcacg ttttcccagg tcagaagcgg ttttcgggag tagtgcccca
      301 actggggtaa cctttgagtt ctctcagttg ggggcgtagg gtcgccgaca tgacacaagg
      361 ggttgtgacc ggggtggaca cgtacgcggg tgcttacgac cgtcagtcgc gcgagcgcga
      421 ctagtacaag ccgaattctg cagatggcgc gccaacctat gcttatccgt ttttatttgt
      481 gtaatggccg cagaattctc tctatcactg atagggaagt ctctatcact gatagggacc
      541 tctctatcac tgatagggaa atctctatca ctgataggga tctctctatc actgataggg
      601 acttctctat cactgatagg gacgtctcta tcactgatag ggaactctct atcactgata
      661 gggacatctc tatcactgat agggacttct ctatcactga tagggaagta tgttctctct
      721 cttctcttct ctctctcttt ctcgaatgtt ctctctcttc tcttctctct ctctttctcg
      781 atggccgggg cgcgccaggt ttcgactttc acttttctct atcactgata gggagatccg
      841 agctcgtaaa ctcgactttc acttttctct atcactgata gggagtggta aactcgaagc
      901 gccggatcca aggtttatgt gattcatata aatacggccg cacaaactct cttcggcatt
      961 agtacaatac aaacaatcaa agtgaaaaca ttaaaaagaa ataacaagcg atttaaatta
     1021 agaaaaacta cgttcaagtg aacaagaaat taaataaatc agaaagtgta caaaagaaag
     1081 taaaataatt ctataaaaag tgaattttaa gcaacaagtt ctataaatag aaaaactatt
     1141 acaaatcatt catcaagttt atatacacaa caaagaagaa aaacaaagtc acacaaatga
     1201 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggac aagaagtaca
     1261 gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     1321 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     1381 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     1441 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     1501 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     1561 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     1621 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     1681 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     1741 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     1801 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     1861 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     1921 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc
     1981 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     2041 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     2101 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     2161 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     2221 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     2281 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     2341 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     2401 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     2461 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     2521 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     2581 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     2641 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     2701 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     2761 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     2821 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     2881 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     2941 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     3001 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     3061 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     3121 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     3181 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     3241 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     3301 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     3361 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     3421 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     3481 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     3541 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     3601 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     3661 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     3721 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccacatcgtg cctcagagct
     3781 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag gcccggggca
     3841 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     3901 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     3961 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     4021 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     4081 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     4141 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     4201 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     4261 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     4321 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     4381 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     4441 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     4501 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     4561 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     4621 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     4681 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     4741 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     4801 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     4861 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     4921 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     4981 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     5041 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     5101 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     5161 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     5221 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     5281 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     5341 gaggcgacag cgctggagga ggtggaagcg gaggaggagg aagcggagga ggaggtagcg
     5401 gacctaagaa aaagaggaag gtgtaaatag aatgaatcgt ttttaaaata acaaatcaat
     5461 tgttttataa tattcgtacg attctttgat tatgtaataa aatgtgatca ttaggaagat
     5521 tacgaaaaat ataaaaaata tgagttctgt gtgtataaca aatgctgtaa acgccacaat
     5581 tgtgtttgtt gcaaataaac ccatgattat ttgattaaaa ttgttgtttt ctttgttcat
     5641 agacaatagt gtgttttgcc taaacgtgta ctgcataaac tccatgcgag tgtatagcga
     5701 gctagtggct aacgcttgcc ccaccaaagt agattcgtca aaatcctcaa tttcatcacc
     5761 ctcctccaag tttaacattt ggccgtcgga attaacttct aaagatgcca cataatctaa
     5821 taaatgaaat agagattcaa acgtggcgtc atcgtccgtt tcgaccattt ccgaaaagaa
     5881 ctcgggcata aactctatga tttctctgga cgtggtgttg tcgaaactct caaagtacgc
     5941 agtcaggaac gtgcgcgaca tgtcgtcggg aaactcgcgc ggaaacatgt tgttgtaacc
     6001 gaacgggtcc catagcgcca aaaccaaatc tgccagcgtc aatagaatga gcacgatgcc
     6061 gacaatggag ctggcttgga tagcgattcg agttaactac tgtccgtttt tatttgtgta
     6121 ataaaatgaa cggaacgctg aattctggaa cattagaaat tctttgttat agaatctata
     6181 aatttatgtc aaagttttgt cgtttcttat gcgatgagag atcttaaaag gcaagtaaat
     6241 tgagctgtcc tattcaaaca tttttctatt aaattgttgg taccttccgc gtcgacgatt
     6301 caatatcgag caaaaatacg aacgcatatg attccatgaa acaaatgtat ttctttagac
     6361 gtttctaaaa ccaattccga caattatttg attctatgac aactccaaat agttttgttt
     6421 attaaaaaat aaaatttcaa caaaaaaagc accgactcgg tgccactttt tcaagttgat
     6481 aacggactag ccttatttta acttgctatt tctagctcta aaacaacctt tatttgatcg
     6541 cgattgcacc agccgggaat cgaacccggg tctgtaccgt ggcagggtac tattctacca
     6601 ctagaccact ggtgctttgt tagcaccgac tcggtgccac tttttcaagt tgataacgga
     6661 ctagccttat tttaacttgc tatttctagc tctaaaactt acaacaggat tggcagcatg
     6721 caccagccgg gaatcgaacc cgggtctgta ccgtggcagg gtactattct accactagac
     6781 cactggtgct ttgttagcac cgactcggtg ccactttttc aagttgataa cggactagcc
     6841 ttattttaac ttgctatttc tagctctaaa acgtttacag tcgattttaa gttgcatcgg
     6901 ccgggaatcg aacccgggcc gcccgcgtgg caggcgagca ttctaccact gaaccaccga
     6961 tgcttgatgt ctacacacac tcaaagccca aagatcatca ctttaataaa ctatttaacc
     7021 gttctttttg agaaaaaaga atatgggaat tataaacaac aatactatat gttgaaaaca
     7081 taaaaatatc atttacagat tagggtttgt tattatcaca ttattactac ctgagtcgtc
     7141 gacgccggat ccttcaagtt cctcttgtac ccatcaaaat gaggccaaac tagtgctgtt
     7201 tttaatgcac tcactatgca tttttatagc ttcctctaca cctcggcaag ttcattaaac
     7261 ttttgccaag cattttaaca attactattt ttagttgttt tccctctttt tccccccatt
     7321 aagtaataaa acattaaaaa attttctttt ggaatatttt cattcaatgc agcttaagcc
     7381 tttcatttta ttaggttctt gtttttttcg tgtttttaaa gtgcacttct ttaaaatttg
     7441 ttcttttcat tcatgttttt tcaaatttga aaataaaaaa aatttacaag tgttttgttt
     7501 tggattcgtt atcattttta aaataacagt ttgtttaaaa aaaagtatat tgaatgttta
     7561 gttgagtagt ttcagctaat aggtcgagaa ttttttcaat acacacttct tctgtctttt
     7621 ttttggtttt gatggttgtc gatgatgggt ttggctgcaa gcttttattg aaacaagtgg
     7681 tgacgggctt cggcacgttc gtattgttct actacggtgt aatcttcatt atgactggta
     7741 atatccaatt ttgaatcgac gtagtagtaa cctggcaact taacgggctt tttggccatg
     7801 tagatggact taaattcaac caagtagtga ccaccaccct tcaatttcaa agccttatga
     7861 atttcacctt tcaagacacc atcacgggga tacaaacgtt cagtagaagg ttcccaaccc
     7921 aaggttttct tttgcataac aagaccatct gatggaaagt ttacaccgat gaatttgacg
     7981 tggtaaataa aagtaccatc ttgcaaagag gaatcttgag taacggtgac aacaccacca
     8041 tcttcgaaat tcattacacg ttcccactta aaaccttcgg ggaaggacaa ctttttataa
     8101 tcgggaatat cagcgggatg cttaacgtat accttagaac cgtattggaa ttgtggggac
     8161 aaaatatccc aggcgaatgg caagggacca cccttggtaa cttgcaattt agcagtttga
     8221 gtaccttcgt agggtttacc ttcaccttca ccttcgattt caaattcgtg accattgaca
     8281 gaaccttcca tgtgtacctt aaaacgcata aaaggcttga taacattttc agtagaatcc
     8341 atggatcctt taagctgagc aatttcagct tggaaagcaa aagtttcaac ttcttcaggc
     8401 atctgtgttt aataaaaaaa aaatagaaaa atacaaatta ttagtaaaat aatcaatcaa
     8461 aatatgaagt ataaataatt ttaatgacgc acgtttttcg ttaagtgtag ggagtgggat
     8521 tggtactgaa aatttttaat aaaaatctag aaatctcttt aagttaagaa ttactagttt
     8581 tttaggggtg gattgggtat gctttttaac gaatgaaaac atgaaatatc gattataatt
     8641 tcgaaaaaat tttgaaaaat tttctttaaa aaatcgcatg ctagaaattt ctttttatta
     8701 gcatggtgaa attttcttta tatactagat ttattttcta aaaatgtcat ggccaaagaa
     8761 aatttaatct agaacaattt acaattactt agtaacgtta ttcttaaacc atattgaagg
     8821 tttttctttc tggcaaatta cttgagtttt tacatcacac ttttttcaaa agaacaaaaa
     8881 cgttttgtta ctacatttta ttttttttaa taaaaactta ccttgatgtg tttgttttat
     8941 attaatttca caaaaataaa tactttttaa attatttttc tttttgcaat tgaaattaaa
     9001 caactacaca cttttttaat gttctttttt cagacttttc tttattaaga aaattaaaac
     9061 acttttaact ttctcccgtt tggaattctt ggtgtttttt gcacaatttt tctttcaaga
     9121 ctgatgccgg ttagcagcca tgcactgaat ttataggtaa aacaaaaata aacgaaaata
     9181 ctagaatgtt ctggaacaaa ctacccgtac ttccgtaggc tctaaacaac atcatgtggt
     9241 tgtttatttg gtgataacaa aagaaattgg ttacatttta gatatataaa aaaattaatt
     9301 tattaaaatt gtagaaaacc aatattaatt ataggttttt attgttattt taatttatta
     9361 gttgcaaatt atcaaaaaaa tgtgcaaaac attcaacatg attaatgccc acacagcgtt
     9421 gtatatatgt agaagcagtg gcttttaaac tgtaaaattg catcaatgtc aaaatttcta
     9481 caatgttctc tctgttttta ataacatgcg gaatgaatca actataattt actttattta
     9541 tttatgcaaa aatatataaa agttattaat taaatgaaaa aggaattaat cttaggcatt
     9601 gctgagtaat gtcatgacat aaactaaata tttttctagt caaacaatga acttatttat
     9661 ttcaaaaacg ttttattctt taaattatag ttaaaaagtc ttattaattg acaaatctat
     9721 gtgtaattaa gtgttttttg aactaagtta caagcttaat atgtatttga ttaattgaaa
     9781 tatgtattaa ctattaattt ttaaattaaa taaatttaat attaattata tattaaactt
     9841 tgtgctttat atggaaatag tatgtatgat ttaatcacat tcgatttaca tattagtcat
     9901 cataaattat aaatgaaagt aatttaattg ttttattaaa gaaaataata acattattaa
     9961 aacaataata ttgctatcaa aatttataaa aatgcataaa atgaaaaaat attgattttc
    10021 tctcatcatg ggggcattaa taattttttc gccagtgcag ctgatgtgca gtaaaacatc
    10081 agagttgccg gtcttgaata aaaacaatga aaagtggaat aatttaaaaa tgtgacaatc
    10141 atatactttt aacatatttt ataaaatttg actgtaaaaa tttaaaaatt aaaatttgca
    10201 aacattatct catacatatt ataacaactc ttttgttttt atataaattt aataaatgaa
    10261 atcaaaaatt ctctcacaaa tttgtaattt ccaaaagtgt ggccttttat tttccccttt
    10321 ttaggtaaaa atttcaaata tctacaaccc tgtaaatgac agctgtttgc gctcttttat
    10381 gtattttttt tgtttatcat gtcggttttt tagttcgcgg tgcaatttac tcgaaatttc
    10441 ttaaagtttg ttgaattttt ggcggtataa ttataatatt ttgttttagt gttcttttta
    10501 tcaaattata atgtttaagt gaattatttt ctgataatgg tttcaacacg tgggcccatc
    10561 tgcagaattc ggcttgtact agtcgcgctc gcgcgactga cggtcgtaag cacccgcgta
    10621 cgtgtccacc ccggtcacaa ccccttgtgt catgtcggcg accctacgcc cccaactgag
    10681 agaactcaaa ggttacccca gttggggcac tactcccgaa aaccgcttct gacctgggaa
    10741 aacgtgaagc cccggggcat ccgctgaggg ttgccgccgg ggcttcggtg tgtccgtcag
    10801 tgagctcctc aggtaataat aacaactaca accaaaaaac aaaagcaaca acaacgacaa
    10861 cttctacagg gcaagtaaca aaacatagta tttgtcaacg acacactata tgaatgatga
    10921 tgatgatgca acaacagcaa caacagcaac aacagctcga gtatcaacaa cacaacaaaa
    10981 aaaacttttt taaaaagaca cagctgctgg caacaaaata ccacaaaaga tataaactaa
    11041 cgcagttgcc acacacacac atacatacat aaattaaaat attatcaaac acacacatat
    11101 acatatatca caacatacct atatacaaaa ccacatttac tttgataaaa ggcgattcaa
    11161 caaaaaagta gtcgtcatcc tccttcattt atccttgcat ttcacgtgta tctgtgtgtg
    11221 ttataaaacc tgctggccaa aaggcaggta ggtagttgac aacacaaata ttttacttct
    11281 ttttcatcat cagcatcaac aacagcagga gcaacatatt gtagacacac actctctgtt
    11341 tcttcacttc acacacatac aagtaaatta tgacagtgaa aaagacgggt atttccaagc
    11401 ttcagtttcg tttactttag gtcattaaga tgactgtcct gatagcttga tatcaatgga
    11461 atcaacggga ttgataaatg gaattactgt gattgataaa tagcttcgaa agacctcgtt
    11521 acttcagggt acttaacaaa atttactgtt ggccgcgaac cgcgccatgg cgccggccgg
    11581 gagccggcgc atgcgaaggt cgcctatggc attattgtac ggaatgataa acattgcctg
    11641 cataaattct tttattatat acagccataa tgtcagtagc aacggagaaa aggttcaaag
    11701 tcgcaaaaaa tttatgagaa acctttacat gagcctgacg tcatcgttta tgcgtaagcg
    11761 tttagaagct cctactttga agagatattt gcgcgataat atctctaata ttttgccaaa
    11821 tgaagtgcct ggtacatcag atgacagtac tgaagagcca gtaatgaaaa aacgtactta
    11881 ctgtacttac tgcccctcta aaataaggcg aaaggcaaat gcatcgtgca aaaaatgcaa
    11941 aaaagttatt tgtcgagagc ataatattga tatgtgccaa agttgtttct gactgactaa
    12001 taagtataat ttgtttctat tatgtataag ttaagctaat tacttatttt ataatacaac
    12061 atgactgttt ttaaagtaca aaataagttt atttttgtaa aagagagaat gtttaaaagt
    12121 tttgttactt tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat
    12181 aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat
    12241 atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt
    12301 ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg gttaagggcc
    12361 cgggatccga tatctagatg cattcgcgag gtaccgagct cgaattcact ggccgtcgtt
    12421 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
    12481 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
    12541 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
    12601 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
    12661 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
    12721 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
    12781 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
    12841 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
    12901 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
    12961 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
    13021 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
    13081 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
    13141 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
    13201 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
    13261 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
    13321 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
    13381 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
    13441 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
    13501 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
    13561 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
    13621 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
    13681 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
    13741 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
    13801 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
    13861 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
    13921 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
    13981 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
    14041 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
    14101 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
    14161 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
    14221 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
    14281 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
    14341 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
    14401 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
    14461 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
    14521 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
    14581 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
    14641 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
    14701 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
    14761 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
    14821 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
    14881 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
    14941 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
    15001 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcag gcctctgcag
    15061 tcgacgggcc caagcttcct ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc
    15121 attcttgaaa tattgctctc tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga
    15181 catctcagtc gccgcttgga gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact
    15241 gattttgaac tataacgacc gcgtgagtca aaatgacgca tgattatctt ttacgtgact
    15301 tttaagattt aactcatacg ataattatat tgttatttca tgttctactt acgtgataac
    15361 ttattatata tatattttct tgttatagat atcgtgacta atatataata aaatgggtag
    15421 ttctttagac gatgagcata tcctctctgc tcttctgcaa agcgatgacg agcttgttgg
    15481 tgaggattct gacagtgaaa tatcagatca cgtaagtgaa gatgacgtcc agagcgatac
    15541 agaagaagcg tttatagatg aggtacatga agtgcagcca acgtcaagcg gtagtgaaat
    15601 attagacgaa caaaatgtta ttgaacaacc aggttcttca ttggcttcta acagaatctt
    15661 gaccttgcga cagaggacta ttagaggtaa gaataaacat tgttggtcaa cttcaaagtc
    15721 cacgaggcgt agcggagtgg taccaggccc tcgagtctgg gcccatgcat gggagctctc
    15781 ccaagatctt atggtcgacc tgcaggc
//
