LOCUS       sequence-494614-       16087 bp    DNA     circular SYN 04-DEC-2025
DEFINITION  Expresses Cas9 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 16087)
  TITLE     bioRxiv -- Conditional expression of Cas9 and dCas9 in Lucilia 
            cuprina reveals dCas9-associated lethality
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 16087)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Cas9-tra135
FEATURES             Location/Qualifiers
     source          1..16087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        complement(300..375)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(474..549)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(648..723)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    4489..4588
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     repeat_region   6139..6201
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6258..6275)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6258..6274)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6267..6289)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6483..6502)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6602..6624
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6662..6680)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6748..6852
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6853..6921,6922..7713)
                     /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(7071..7090)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7884..8472
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8373..8392
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8626..8643
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8760..8781
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8796..8801,8802..8819,8820..8826)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8834..8850
                     /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     8839..8861
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8858..8874
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8858..8874
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   8935..8969
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    9922..10021
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     protein_bind    10156..10174
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10177..10195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10198..10216
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10219..10237
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10240..10258
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10261..10279
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10282..10300
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10303..10321
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10431..10449
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10473..10491
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10515..10533
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10565..10583
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10607..10625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10649..10667
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10691..10709
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10741..10759
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10783..10801
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10825..10843
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             11169..11189
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11214..15314
                     /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="DKKYSIGLDIGTNSVGWAVITDEYEVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             15315..15362
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 acattagaaa ttctttgtta tagaatctat aaatttatgt caaagttttg tcgtttctta
       61 tgcgatgaga gatcttaaaa ggcaagtaaa ttgagctgtc ctattcaaac atttttctat
      121 taaattgttg gtaccttccg cgtcgacgat tcaatatcga gcaaaaatac gaacgcatat
      181 gattccatga aacaaatgta tttctttaga cgtttctaaa accaattccg acaattattt
      241 gattctatga caactccaaa tagttttgtt tattaaaaaa taaaatttca acaaaaaaag
      301 caccgactcg gtgccacttt ttcaagttga taacggacta gccttatttt aacttgctat
      361 ttctagctct aaaactcacc taaacataga gataatgcac cagccgggaa tcgaacccgg
      421 gtctgtaccg tggcagggta ctattctacc actagaccac tggtgctttg ttagcaccga
      481 ctcggtgcca ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag
      541 ctctaaaaca tgataagcag ttgttgtggt gcaccagccg ggaatcgaac ccgggtctgt
      601 accgtggcag ggtactattc taccactaga ccactggtgc tttgttagca ccgactcggt
      661 gccacttttt caagttgata acggactagc cttattttaa cttgctattt ctagctctaa
      721 aacgctaagt tttcaactac gtatgcatcg gccgggaatc gaacccgggc cgcccgcgtg
      781 gcaggcgagc attctaccac tgaaccaccg atgcttgatg tctacacaca ctcaaagccc
      841 aaagatcatc actttaataa actatttaac cgttcttttt gagaaaaaag aatatgggaa
      901 ttataaacaa caatactata tgttgaaaac ataaaaatat catttacaga ttagggtttg
      961 ttattatcac attattacta cctgagtcgt cgacgccgat ccttcaagtt cctcttgtac
     1021 ccatcaaaat gaggccaaac tagtgctgtt tttaatgcac tcactatgca tttttatagc
     1081 ttcctctaca cctcggcaag ttcattaaac ttttgccaag cattttaaca attactattt
     1141 ttagttgttt tccctctttt tccccccatt aagtaataaa acattaaaaa attttctttt
     1201 ggaatatttt cattcaatgc agcttaagcc tttcatttta ttaggttctt gtttttttcg
     1261 tgtttttaaa gtgcacttct ttaaaatttg ttcttttcat tcatgttttt tcaaatttga
     1321 aaataaaaaa aatttacaag tgttttgttt tggattcgtt atcattttta aaataacagt
     1381 ttgtttaaaa aaaagtatat tgaatgttta gttgagtagt ttcagctaat aggtcgagaa
     1441 ttttttcaat acacacttct tctgtctttt ttttggtttt gatggttgtc gatgatgggt
     1501 ttggctgcaa gcttttattg aaacaagtgg tgacgggctt cggcacgttc gtattgttct
     1561 actacggtgt aatcttcatt atgactggta atatccaatt ttgaatcgac gtagtagtaa
     1621 cctggcaact taacgggctt tttggccatg tagatggact taaattcaac caagtagtga
     1681 ccaccaccct tcaatttcaa agccttatga atttcacctt tcaagacacc atcacgggga
     1741 tacaaacgtt cagtagaagg ttcccaaccc aaggttttct tttgcataac aagaccatct
     1801 gatggaaagt ttacaccgat gaatttgacg tggtaaataa aagtaccatc ttgcaaagag
     1861 gaatcttgag taacggtgac aacaccacca tcttcgaaat tcattacacg ttcccactta
     1921 aaaccttcgg ggaaggacaa ctttttataa tcgggaatat cagcgggatg cttaacgtat
     1981 accttagaac cgtattggaa ttgtggggac aaaatatccc aggcgaatgg caagggacca
     2041 cccttggtaa cttgcaattt agcagtttga gtaccttcgt agggtttacc ttcaccttca
     2101 ccttcgattt caaattcgtg accattgaca gaaccttcca tgtgtacctt aaaacgcata
     2161 aaaggcttga taacattttc agtagaatcc atggatcctt taagctgagc aatttcagct
     2221 tggaaagcaa aagtttcaac ttcttcaggc atctgtgttt aataaaaaaa aaatagaaaa
     2281 atacaaatta ttagtaaaat aatcaatcaa aatatgaagt ataaataatt ttaatgacgc
     2341 acgtttttcg ttaagtgtag ggagtgggat tggtactgaa aatttttaat aaaaatctag
     2401 aaatctcttt aagttaagaa ttactagttt tttaggggtg gattgggtat gctttttaac
     2461 gaatgaaaac atgaaatatc gattataatt tcgaaaaaat tttgaaaaat tttctttaaa
     2521 aaatcgcatg ctagaaattt ctttttatta gcatggtgaa attttcttta tatactagat
     2581 ttattttcta aaaatgtcat ggccaaagaa aatttaatct agaacaattt acaattactt
     2641 agtaacgtta ttcttaaacc atattgaagg tttttctttc tggcaaatta cttgagtttt
     2701 tacatcacac ttttttcaaa agaacaaaaa cgttttgtta ctacatttta ttttttttaa
     2761 taaaaactta ccttgatgtg tttgttttat attaatttca caaaaataaa tactttttaa
     2821 attatttttc tttttgcaat tgaaattaaa caactacaca cttttttaat gttctttttt
     2881 cagacttttc tttattaaga aaattaaaac acttttaact ttctcccgtt tggaattctt
     2941 ggtgtttttt gcacaatttt tctttcaaga ctgatgccgg ttagcagcca tgcactgaat
     3001 ttataggtaa aacaaaaata aacgaaaata ctagaatgtt ctggaacaaa ctacccgtac
     3061 ttccgtaggc tctaaacaac atcatgtggt tgtttatttg gtgataacaa aagaaattgg
     3121 ttacatttta gatatataaa aaaattaatt tattaaaatt gtagaaaacc aatattaatt
     3181 ataggttttt attgttattt taatttatta gttgcaaatt atcaaaaaaa tgtgcaaaac
     3241 attcaacatg attaatgccc acacagcgtt gtatatatgt agaagcagtg gcttttaaac
     3301 tgtaaaattg catcaatgtc aaaatttcta caatgttctc tctgttttta ataacatgcg
     3361 gaatgaatca actataattt actttattta tttatgcaaa aatatataaa agttattaat
     3421 taaatgaaaa aggaattaat cttaggcatt gctgagtaat gtcatgacat aaactaaata
     3481 tttttctagt caaacaatga acttatttat ttcaaaaacg ttttattctt taaattatag
     3541 ttaaaaagtc ttattaattg acaaatctat gtgtaattaa gtgttttttg aactaagtta
     3601 caagcttaat atgtatttga ttaattgaaa tatgtattaa ctattaattt ttaaattaaa
     3661 taaatttaat attaattata tattaaactt tgtgctttat atggaaatag tatgtatgat
     3721 ttaatcacat tcgatttaca tattagtcat cataaattat aaatgaaagt aatttaattg
     3781 ttttattaaa gaaaataata acattattaa aacaataata ttgctatcaa aatttataaa
     3841 aatgcataaa atgaaaaaat attgattttc tctcatcatg ggggcattaa taattttttc
     3901 gccagtgcag ctgatgtgca gtaaaacatc agagttgccg gtcttgaata aaaacaatga
     3961 aaagtggaat aatttaaaaa tgtgacaatc atatactttt aacatatttt ataaaatttg
     4021 actgtaaaaa tttaaaaatt aaaatttgca aacattatct catacatatt ataacaactc
     4081 ttttgttttt atataaattt aataaatgaa atcaaaaatt ctctcacaaa tttgtaattt
     4141 ccaaaagtgt ggccttttat tttccccttt ttaggtaaaa atttcaaata tctacaaccc
     4201 tgtaaatgac agctgtttgc gctcttttat gtattttttt tgtttatcat gtcggttttt
     4261 tagttcgcgg tgcaatttac tcgaaatttc ttaaagtttg ttgaattttt ggcggtataa
     4321 ttataatatt ttgttttagt gttcttttta tcaaattata atgtttaagt gaattatttt
     4381 ctgataatgg tttcaacacg tgggcccatc tgcagaattc ggcttgtact agtcgcgctc
     4441 gcgcgactga cggtcgtaag cacccgcgta cgtgtccacc ccggtcacaa ccccttgtgt
     4501 catgtcggcg accctacgcc cccaactgag agaactcaaa ggttacccca gttggggcac
     4561 tactcccgaa aaccgcttct gacctgggaa aacgtgaagc cccggggcat ccgctgaggg
     4621 ttgccgccgg ggcttcggtg tgtccgtcag tgagctcctc aggtaataat aacaactaca
     4681 accaaaaaac aaaagcaaca acaacgacaa cttctacagg gcaagtaaca aaacatagta
     4741 tttgtcaacg acacactata tgaatgatga tgatgatgca acaacagcaa caacagcaac
     4801 aacagctcga gtatcaacaa cacaacaaaa aaaacttttt taaaaagaca cagctgctgg
     4861 caacaaaata ccacaaaaga tataaactaa cgcagttgcc acacacacac atacatacat
     4921 aaattaaaat attatcaaac acacacatat acatatatca caacatacct atatacaaaa
     4981 ccacatttac tttgataaaa ggcgattcaa caaaaaagta gtcgtcatcc tccttcattt
     5041 atccttgcat ttcacgtgta tctgtgtgtg ttataaaacc tgctggccaa aaggcaggta
     5101 ggtagttgac aacacaaata ttttacttct ttttcatcat cagcatcaac aacagcagga
     5161 gcaacatatt gtagacacac actctctgtt tcttcacttc acacacatac aagtaaatta
     5221 tgacagtgaa aaagacgggt atttccaagc ttcagtttcg tttactttag gtcattaaga
     5281 tgactgtcct gatagcttga tatcaatgga atcaacggga ttgataaatg gaattactgt
     5341 gattgataaa tagcttcgaa agacctcgtt acttcagggt acttaacaaa atttactgtt
     5401 ggccgcgaac cgcgccatgg cgccggccgg gagccggcgc atgcgaaggt cgcctatggc
     5461 attattgtac ggaatgataa acattgcctg cataaattct tttattatat acagccataa
     5521 tgtcagtagc aacggagaaa aggttcaaag tcgcaaaaaa tttatgagaa acctttacat
     5581 gagcctgacg tcatcgttta tgcgtaagcg tttagaagct cctactttga agagatattt
     5641 gcgcgataat atctctaata ttttgccaaa tgaagtgcct ggtacatcag atgacagtac
     5701 tgaagagcca gtaatgaaaa aacgtactta ctgtacttac tgcccctcta aaataaggcg
     5761 aaaggcaaat gcatcgtgca aaaaatgcaa aaaagttatt tgtcgagagc ataatattga
     5821 tatgtgccaa agttgtttct gactgactaa taagtataat ttgtttctat tatgtataag
     5881 ttaagctaat tacttatttt ataatacaac atgactgttt ttaaagtaca aaataagttt
     5941 atttttgtaa aagagagaat gtttaaaagt tttgttactt tatagaagaa attttgagtt
     6001 tttgtttttt tttaataaat aaataaacat aaataaattg tttgttgaat ttattattag
     6061 tatgtaagtg taaatataat aaaacttaat atctattcaa attaataaat aaacctcgat
     6121 atacagaccg ataaaacaca tgcgtcaatt ttacgcatga ttatctttaa cgtacgtcac
     6181 aatatgatta tctttctagg gttaagggcc cgggatccga tatctagatg cattcgcgag
     6241 gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     6301 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     6361 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     6421 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     6481 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     6541 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6601 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     6661 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6721 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6781 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6841 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6901 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6961 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     7021 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     7081 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     7141 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     7201 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7261 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7321 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7381 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7441 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7501 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7561 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7621 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7681 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7741 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7801 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7861 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7921 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7981 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     8041 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     8101 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     8161 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     8221 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     8281 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     8341 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8401 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8461 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8521 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     8581 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     8641 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     8701 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     8761 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     8821 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     8881 tacgccaagc ttgcatgcag gcctctgcag tcgacgggcc caagcttcct ttaaccctag
     8941 aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc tctttctaaa
     9001 tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga gctcccgtga
     9061 ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc gcgtgagtca
     9121 aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg ataattatat
     9181 tgttatttca tgttctactt acgtgataac ttattatata tatattttct tgttatagat
     9241 atcgtgacta atatataata aaatgggtag ttctttagac gatgagcata tcctctctgc
     9301 tcttctgcaa agcgatgacg agcttgttgg tgaggattct gacagtgaaa tatcagatca
     9361 cgtaagtgaa gatgacgtcc agagcgatac agaagaagcg tttatagatg aggtacatga
     9421 agtgcagcca acgtcaagcg gtagtgaaat attagacgaa caaaatgtta ttgaacaacc
     9481 aggttcttca ttggcttcta acagaatctt gaccttgcga cagaggacta ttagaggtaa
     9541 gaataaacat tgttggtcaa cttcaaagtc cacgaggcgt agcggagtgg taccaggccc
     9601 tcgagtctgg gcccatgcat gggagctctc ccaagatctt atggtcgacc tgcaggcaac
     9661 attcatgtgt gggtgattac actcacacaa aaacacacac attcacacaa acacacacac
     9721 ttataaagat gagaacattt aacaaaaaca attataaaac tgatggcaat ggagcaatga
     9781 tgtgatgaag ctgatgaaag ttttgaatat tcattcaaga gttaaaaaca tgaacaaaca
     9841 tgcacaaaat tagtcgagac tgacggacac accgaagccc cggcggcaac cctcagcgga
     9901 tgccccgggg cttcacgttt tcccaggtca gaagcggttt tcgggagtag tgccccaact
     9961 ggggtaacct ttgagttctc tcagttgggg gcgtagggtc gccgacatga cacaaggggt
    10021 tgtgaccggg gtggacacgt acgcgggtgc ttacgaccgt cagtcgcgcg agcgcgacta
    10081 gtacaagccg aattctgcag atggcgcgcc aacctatgct tatccgtttt tatttgtgta
    10141 atggccgcag aattctctct atcactgata gggaggtctc tatcactgat agggagttct
    10201 ctatcactga tagggatgtc tctatcactg atagggacgt ctctatcact gatagggaac
    10261 tctctatcac tgatagggac atctctatca ctgataggga cttctctatc actgataggg
    10321 aagtatgttc tctctcttct cttctctctc tctttctcga atgttctctc tcttctcttc
    10381 tctctctctt tctcgatggc cggggcgcgc caggtttcga ctttcacttt tctctatcac
    10441 tgatagggag tggtaaactc gactttcact tttctctatc actgataggg agtggtaaac
    10501 tcgactttca cttttctcta tcactgatag ggagatccga gctcgtaaac tcgactttca
    10561 cttttctcta tcactgatag ggagtggtaa actcgacttt cacttttctc tatcactgat
    10621 agggagtggt aaactcgact ttcacttttc tctatcactg atagggagtg gtaaactcga
    10681 ctttcacttt tctctatcac tgatagggag atccgagctc gtaaactcga ctttcacttt
    10741 tctctatcac tgatagggag tggtaaactc gactttcact tttctctatc actgataggg
    10801 agtggtaaac tcgactttca cttttctcta tcactgatag ggagtggtaa actcgaagcg
    10861 ccggatccaa ggtttatgtg attcatataa atacggccgc acaaactctc ttcggcatta
    10921 gtacaataca aacaatcaaa gtgaaaacat taaaaagaaa taacaagcga tttaaattaa
    10981 gaaaaactac gttcaagtga acaagaaatt aaataaatca gaaagtgtac aaaagaaagt
    11041 aaaataattc tataaaaagt gaattttaag caacaagttc tataaataga aaaactatta
    11101 caaatcattc atcaagttta tatacacaac aaagaagaaa aacaaagtca cacaaatgaa
    11161 aaggcatgcc aaagaagaag cggaaggtcg gtatccacgg agtcccagca gccgacaaga
    11221 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
    11281 agtacgaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
    11341 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
    11401 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
    11461 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
    11521 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
    11581 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
    11641 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
    11701 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
    11761 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
    11821 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
    11881 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc
    11941 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
    12001 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
    12061 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
    12121 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
    12181 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
    12241 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
    12301 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
    12361 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
    12421 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
    12481 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
    12541 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
    12601 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
    12661 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
    12721 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
    12781 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
    12841 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
    12901 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
    12961 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
    13021 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
    13081 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
    13141 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
    13201 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
    13261 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
    13321 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
    13381 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
    13441 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
    13501 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
    13561 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
    13621 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
    13681 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
    13741 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
    13801 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
    13861 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
    13921 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
    13981 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
    14041 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
    14101 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
    14161 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
    14221 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
    14281 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
    14341 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
    14401 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
    14461 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
    14521 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
    14581 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
    14641 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
    14701 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
    14761 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
    14821 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
    14881 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
    14941 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
    15001 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
    15061 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
    15121 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
    15181 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
    15241 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
    15301 agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa
    15361 agtaatagaa tgaatcgttt ttaaaataac aaatcaattg ttttataata ttcgtacgat
    15421 tctttgatta tgtaataaaa tgtgatcatt aggaagatta cgaaaaatat aaaaaatatg
    15481 agttctgtgt gtataacaaa tgctgtaaac gccacaattg tgtttgttgc aaataaaccc
    15541 atgattattt gattaaaatt gttgttttct ttgttcatag acaatagtgt gttttgccta
    15601 aacgtgtact gcataaactc catgcgagtg tatagcgagc tagtggctaa cgcttgcccc
    15661 accaaagtag attcgtcaaa atcctcaatt tcatcaccct cctccaagtt taacatttgg
    15721 ccgtcggaat taacttctaa agatgccaca taatctaata aatgaaatag agattcaaac
    15781 gtggcgtcat cgtccgtttc gaccatttcc gaaaagaact cgggcataaa ctctatgatt
    15841 tctctggacg tggtgttgtc gaaactctca aagtacgcag tcaggaacgt gcgcgacatg
    15901 tcgtcgggaa actcgcgcgg aaacatgttg ttgtaaccga acgggtccca tagcgccaaa
    15961 accaaatctg ccagcgtcaa tagaatgagc acgatgccga caatggagct ggcttggata
    16021 gcgattcgag ttaactactg tccgttttta tttgtgtaat aaaatgaacg gaacgctgaa
    16081 ttctgga
//
