LOCUS       sequence-494616-       16080 bp    DNA     circular SYN 15-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 16080)
  TITLE     bioRxiv -- Conditional expression of Cas9 and dCas9 in Lucilia 
            cuprina reveals dCas9-associated lethality
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 16080)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Cas9-tra246
FEATURES             Location/Qualifiers
     source          1..16080
                     /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    4490..4589
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     repeat_region   6140..6202
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6259..6276)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6259..6275)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6268..6290)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6484..6503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6603..6625
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6663..6681)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6749..6853
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6854..6922,6923..7714)
                     /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(7072..7091)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7885..8473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8374..8393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8627..8644
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8761..8782
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8797..8802,8803..8820,8821..8827)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8835..8851
                     /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     8840..8862
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8859..8875
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8859..8875
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   8936..8970
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    9923..10022
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     protein_bind    10157..10175
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10178..10196
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10199..10217
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10220..10238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10241..10259
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10262..10280
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10283..10301
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10304..10322
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    join(10432..10450,10451..10473,10474..10492,10493..10515,
                     10516..10534,10535..10557,10558..10576,10577..10599,
                     10600..10618,10619..10641,10642..10660,10661..10683,
                     10684..10702)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    10734..10752
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10776..10794
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10818..10836
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             11162..11182
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11207..15307
                     /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             15308..15355
                     /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 aaaacaacct ttatttgatc gcgattgcac cagccgggaa tcgaacccgg
      421 gtctgtaccg tggcagggta ctattctacc actagaccac tggtgctttg ttagcaccga
      481 ctcggtgcca ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag
      541 ctctaaaact tacaacagga ttggcagcat gcaccagccg ggaatcgaac ccgggtctgt
      601 accgtggcag ggtactattc taccactaga ccactggtgc tttgttagca ccgactcggt
      661 gccacttttt caagttgata acggactagc cttattttaa cttgctattt ctagctctaa
      721 aacgtttaca gtcgatttta agttgcatcg 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 cgacgccgga tccttcaagt tcctcttgta
     1021 cccatcaaaa tgaggccaaa ctagtgctgt ttttaatgca ctcactatgc atttttatag
     1081 cttcctctac acctcggcaa gttcattaaa cttttgccaa gcattttaac aattactatt
     1141 tttagttgtt ttccctcttt ttccccccat taagtaataa aacattaaaa aattttcttt
     1201 tggaatattt tcattcaatg cagcttaagc ctttcatttt attaggttct tgtttttttc
     1261 gtgtttttaa agtgcacttc tttaaaattt gttcttttca ttcatgtttt ttcaaatttg
     1321 aaaataaaaa aaatttacaa gtgttttgtt ttggattcgt tatcattttt aaaataacag
     1381 tttgtttaaa aaaaagtata ttgaatgttt agttgagtag tttcagctaa taggtcgaga
     1441 attttttcaa tacacacttc ttctgtcttt tttttggttt tgatggttgt cgatgatggg
     1501 tttggctgca agcttttatt gaaacaagtg gtgacgggct tcggcacgtt cgtattgttc
     1561 tactacggtg taatcttcat tatgactggt aatatccaat tttgaatcga cgtagtagta
     1621 acctggcaac ttaacgggct ttttggccat gtagatggac ttaaattcaa ccaagtagtg
     1681 accaccaccc ttcaatttca aagccttatg aatttcacct ttcaagacac catcacgggg
     1741 atacaaacgt tcagtagaag gttcccaacc caaggttttc ttttgcataa caagaccatc
     1801 tgatggaaag tttacaccga tgaatttgac gtggtaaata aaagtaccat cttgcaaaga
     1861 ggaatcttga gtaacggtga caacaccacc atcttcgaaa ttcattacac gttcccactt
     1921 aaaaccttcg gggaaggaca actttttata atcgggaata tcagcgggat gcttaacgta
     1981 taccttagaa ccgtattgga attgtgggga caaaatatcc caggcgaatg gcaagggacc
     2041 acccttggta acttgcaatt tagcagtttg agtaccttcg tagggtttac cttcaccttc
     2101 accttcgatt tcaaattcgt gaccattgac agaaccttcc atgtgtacct taaaacgcat
     2161 aaaaggcttg ataacatttt cagtagaatc catggatcct ttaagctgag caatttcagc
     2221 ttggaaagca aaagtttcaa cttcttcagg catctgtgtt taataaaaaa aaaatagaaa
     2281 aatacaaatt attagtaaaa taatcaatca aaatatgaag tataaataat tttaatgacg
     2341 cacgtttttc gttaagtgta gggagtggga ttggtactga aaatttttaa taaaaatcta
     2401 gaaatctctt taagttaaga attactagtt ttttaggggt ggattgggta tgctttttaa
     2461 cgaatgaaaa catgaaatat cgattataat ttcgaaaaaa ttttgaaaaa ttttctttaa
     2521 aaaatcgcat gctagaaatt tctttttatt agcatggtga aattttcttt atatactaga
     2581 tttattttct aaaaatgtca tggccaaaga aaatttaatc tagaacaatt tacaattact
     2641 tagtaacgtt attcttaaac catattgaag gtttttcttt ctggcaaatt acttgagttt
     2701 ttacatcaca cttttttcaa aagaacaaaa acgttttgtt actacatttt atttttttta
     2761 ataaaaactt accttgatgt gtttgtttta tattaatttc acaaaaataa atacttttta
     2821 aattattttt ctttttgcaa ttgaaattaa acaactacac acttttttaa tgttcttttt
     2881 tcagactttt ctttattaag aaaattaaaa cacttttaac tttctcccgt ttggaattct
     2941 tggtgttttt tgcacaattt ttctttcaag actgatgccg gttagcagcc atgcactgaa
     3001 tttataggta aaacaaaaat aaacgaaaat actagaatgt tctggaacaa actacccgta
     3061 cttccgtagg ctctaaacaa catcatgtgg ttgtttattt ggtgataaca aaagaaattg
     3121 gttacatttt agatatataa aaaaattaat ttattaaaat tgtagaaaac caatattaat
     3181 tataggtttt tattgttatt ttaatttatt agttgcaaat tatcaaaaaa atgtgcaaaa
     3241 cattcaacat gattaatgcc cacacagcgt tgtatatatg tagaagcagt ggcttttaaa
     3301 ctgtaaaatt gcatcaatgt caaaatttct acaatgttct ctctgttttt aataacatgc
     3361 ggaatgaatc aactataatt tactttattt atttatgcaa aaatatataa aagttattaa
     3421 ttaaatgaaa aaggaattaa tcttaggcat tgctgagtaa tgtcatgaca taaactaaat
     3481 atttttctag tcaaacaatg aacttattta tttcaaaaac gttttattct ttaaattata
     3541 gttaaaaagt cttattaatt gacaaatcta tgtgtaatta agtgtttttt gaactaagtt
     3601 acaagcttaa tatgtatttg attaattgaa atatgtatta actattaatt tttaaattaa
     3661 ataaatttaa tattaattat atattaaact ttgtgcttta tatggaaata gtatgtatga
     3721 tttaatcaca ttcgatttac atattagtca tcataaatta taaatgaaag taatttaatt
     3781 gttttattaa agaaaataat aacattatta aaacaataat attgctatca aaatttataa
     3841 aaatgcataa aatgaaaaaa tattgatttt ctctcatcat gggggcatta ataatttttt
     3901 cgccagtgca gctgatgtgc agtaaaacat cagagttgcc ggtcttgaat aaaaacaatg
     3961 aaaagtggaa taatttaaaa atgtgacaat catatacttt taacatattt tataaaattt
     4021 gactgtaaaa atttaaaaat taaaatttgc aaacattatc tcatacatat tataacaact
     4081 cttttgtttt tatataaatt taataaatga aatcaaaaat tctctcacaa atttgtaatt
     4141 tccaaaagtg tggcctttta ttttcccctt tttaggtaaa aatttcaaat atctacaacc
     4201 ctgtaaatga cagctgtttg cgctctttta tgtatttttt ttgtttatca tgtcggtttt
     4261 ttagttcgcg gtgcaattta ctcgaaattt cttaaagttt gttgaatttt tggcggtata
     4321 attataatat tttgttttag tgttcttttt atcaaattat aatgtttaag tgaattattt
     4381 tctgataatg gtttcaacac gtgggcccat ctgcagaatt cggcttgtac tagtcgcgct
     4441 cgcgcgactg acggtcgtaa gcacccgcgt acgtgtccac cccggtcaca accccttgtg
     4501 tcatgtcggc gaccctacgc ccccaactga gagaactcaa aggttacccc agttggggca
     4561 ctactcccga aaaccgcttc tgacctggga aaacgtgaag ccccggggca tccgctgagg
     4621 gttgccgccg gggcttcggt gtgtccgtca gtgagctcct caggtaataa taacaactac
     4681 aaccaaaaaa caaaagcaac aacaacgaca acttctacag ggcaagtaac aaaacatagt
     4741 atttgtcaac gacacactat atgaatgatg atgatgatgc aacaacagca acaacagcaa
     4801 caacagctcg agtatcaaca acacaacaaa aaaaactttt ttaaaaagac acagctgctg
     4861 gcaacaaaat accacaaaag atataaacta acgcagttgc cacacacaca catacataca
     4921 taaattaaaa tattatcaaa cacacacata tacatatatc acaacatacc tatatacaaa
     4981 accacattta ctttgataaa aggcgattca acaaaaaagt agtcgtcatc ctccttcatt
     5041 tatccttgca tttcacgtgt atctgtgtgt gttataaaac ctgctggcca aaaggcaggt
     5101 aggtagttga caacacaaat attttacttc tttttcatca tcagcatcaa caacagcagg
     5161 agcaacatat tgtagacaca cactctctgt ttcttcactt cacacacata caagtaaatt
     5221 atgacagtga aaaagacggg tatttccaag cttcagtttc gtttacttta ggtcattaag
     5281 atgactgtcc tgatagcttg atatcaatgg aatcaacggg attgataaat ggaattactg
     5341 tgattgataa atagcttcga aagacctcgt tacttcaggg tacttaacaa aatttactgt
     5401 tggccgcgaa ccgcgccatg gcgccggccg ggagccggcg catgcgaagg tcgcctatgg
     5461 cattattgta cggaatgata aacattgcct gcataaattc ttttattata tacagccata
     5521 atgtcagtag caacggagaa aaggttcaaa gtcgcaaaaa atttatgaga aacctttaca
     5581 tgagcctgac gtcatcgttt atgcgtaagc gtttagaagc tcctactttg aagagatatt
     5641 tgcgcgataa tatctctaat attttgccaa atgaagtgcc tggtacatca gatgacagta
     5701 ctgaagagcc agtaatgaaa aaacgtactt actgtactta ctgcccctct aaaataaggc
     5761 gaaaggcaaa tgcatcgtgc aaaaaatgca aaaaagttat ttgtcgagag cataatattg
     5821 atatgtgcca aagttgtttc tgactgacta ataagtataa tttgtttcta ttatgtataa
     5881 gttaagctaa ttacttattt tataatacaa catgactgtt tttaaagtac aaaataagtt
     5941 tatttttgta aaagagagaa tgtttaaaag ttttgttact ttatagaaga aattttgagt
     6001 ttttgttttt ttttaataaa taaataaaca taaataaatt gtttgttgaa tttattatta
     6061 gtatgtaagt gtaaatataa taaaacttaa tatctattca aattaataaa taaacctcga
     6121 tatacagacc gataaaacac atgcgtcaat tttacgcatg attatcttta acgtacgtca
     6181 caatatgatt atctttctag ggttaagggc ccgggatccg atatctagat gcattcgcga
     6241 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6301 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6361 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     6421 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     6481 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     6541 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     6601 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     6661 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     6721 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6781 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6841 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6901 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6961 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7021 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7081 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7141 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7201 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7261 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7321 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7381 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7441 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7501 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7561 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7621 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7681 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7741 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7801 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7861 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7921 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7981 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     8041 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8101 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8161 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8221 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8281 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8341 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8401 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     8461 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     8521 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     8581 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     8641 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     8701 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     8761 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     8821 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     8881 ttacgccaag cttgcatgca ggcctctgca gtcgacgggc ccaagcttcc tttaacccta
     8941 gaaagatagt ctgcgtaaaa ttgacgcatg cattcttgaa atattgctct ctctttctaa
     9001 atagcgcgaa tccgtcgctg tgcatttagg acatctcagt cgccgcttgg agctcccgtg
     9061 aggcgtgctt gtcaatgcgg taagtgtcac tgattttgaa ctataacgac cgcgtgagtc
     9121 aaaatgacgc atgattatct tttacgtgac ttttaagatt taactcatac gataattata
     9181 ttgttatttc atgttctact tacgtgataa cttattatat atatattttc ttgttataga
     9241 tatcgtgact aatatataat aaaatgggta gttctttaga cgatgagcat atcctctctg
     9301 ctcttctgca aagcgatgac gagcttgttg gtgaggattc tgacagtgaa atatcagatc
     9361 acgtaagtga agatgacgtc cagagcgata cagaagaagc gtttatagat gaggtacatg
     9421 aagtgcagcc aacgtcaagc ggtagtgaaa tattagacga acaaaatgtt attgaacaac
     9481 caggttcttc attggcttct aacagaatct tgaccttgcg acagaggact attagaggta
     9541 agaataaaca ttgttggtca acttcaaagt ccacgaggcg tagcggagtg gtaccaggcc
     9601 ctcgagtctg ggcccatgca tgggagctct cccaagatct tatggtcgac ctgcaggcaa
     9661 cattcatgtg tgggtgatta cactcacaca aaaacacaca cattcacaca aacacacaca
     9721 cttataaaga tgagaacatt taacaaaaac aattataaaa ctgatggcaa tggagcaatg
     9781 atgtgatgaa gctgatgaaa gttttgaata ttcattcaag agttaaaaac atgaacaaac
     9841 atgcacaaaa ttagtcgaga ctgacggaca caccgaagcc ccggcggcaa ccctcagcgg
     9901 atgccccggg gcttcacgtt ttcccaggtc agaagcggtt ttcgggagta gtgccccaac
     9961 tggggtaacc tttgagttct ctcagttggg ggcgtagggt cgccgacatg acacaagggg
    10021 ttgtgaccgg ggtggacacg tacgcgggtg cttacgaccg tcagtcgcgc gagcgcgact
    10081 agtacaagcc gaattctgca gatggcgcgc caacctatgc ttatccgttt ttatttgtgt
    10141 aatggccgca gaattctctc tatcactgat agggaggtct ctatcactga tagggagttc
    10201 tctatcactg atagggatgt ctctatcact gatagggacg tctctatcac tgatagggaa
    10261 ctctctatca ctgataggga catctctatc actgataggg acttctctat cactgatagg
    10321 gaagtatgtt ctctctcttc tcttctctct ctctttctcg aatgttctct ctcttctctt
    10381 ctctctctct ttctcgatgg ccggggcgcg ccaggtttcg actttcactt ttctctatca
    10441 ctgataggga gtggtaaact cgactttcac ttttctctat cactgatagg gagtggtaaa
    10501 ctcgactttc acttttctct atcactgata gggagtggta aactcgactt tcacttttct
    10561 ctatcactga tagggagtgg taaactcgac tttcactttt ctctatcact gatagggagt
    10621 ggtaaactcg actttcactt ttctctatca ctgataggga gtggtaaact cgactttcac
    10681 ttttctctat cactgatagg gagatccgag ctcgtaaact cgactttcac ttttctctat
    10741 cactgatagg gagtggtaaa ctcgactttc acttttctct atcactgata gggagtggta
    10801 aactcgactt tcacttttct ctatcactga tagggagtgg taaactcgaa gcgccggatc
    10861 caaggtttat gtgattcata taaatacggc cgcacaaact ctcttcggca ttagtacaat
    10921 acaaacaatc aaagtgaaaa cattaaaaag aaataacaag cgatttaaat taagaaaaac
    10981 tacgttcaag tgaacaagaa attaaataaa tcagaaagtg tacaaaagaa agtaaaataa
    11041 ttctataaaa agtgaatttt aagcaacaag ttctataaat agaaaaacta ttacaaatca
    11101 ttcatcaagt ttatatacac aacaaagaag aaaaacaaag tcacacaaat gaaaaggcat
    11161 gccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag
    11221 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacga
    11281 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
    11341 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
    11401 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
    11461 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
    11521 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
    11581 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
    11641 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
    11701 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
    11761 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
    11821 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
    11881 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
    11941 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
    12001 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
    12061 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
    12121 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
    12181 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
    12241 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
    12301 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
    12361 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
    12421 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
    12481 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
    12541 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
    12601 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
    12661 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
    12721 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
    12781 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
    12841 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
    12901 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
    12961 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
    13021 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
    13081 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
    13141 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
    13201 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
    13261 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
    13321 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
    13381 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
    13441 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
    13501 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
    13561 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
    13621 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
    13681 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
    13741 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
    13801 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
    13861 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
    13921 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
    13981 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
    14041 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
    14101 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
    14161 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
    14221 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
    14281 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
    14341 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
    14401 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
    14461 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
    14521 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
    14581 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
    14641 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
    14701 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
    14761 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
    14821 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
    14881 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
    14941 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
    15001 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
    15061 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
    15121 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
    15181 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
    15241 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
    15301 aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaagtaata
    15361 gaatgaatcg tttttaaaat aacaaatcaa ttgttttata atattcgtac gattctttga
    15421 ttatgtaata aaatgtgatc attaggaaga ttacgaaaaa tataaaaaat atgagttctg
    15481 tgtgtataac aaatgctgta aacgccacaa ttgtgtttgt tgcaaataaa cccatgatta
    15541 tttgattaaa attgttgttt tctttgttca tagacaatag tgtgttttgc ctaaacgtgt
    15601 actgcataaa ctccatgcga gtgtatagcg agctagtggc taacgcttgc cccaccaaag
    15661 tagattcgtc aaaatcctca atttcatcac cctcctccaa gtttaacatt tggccgtcgg
    15721 aattaacttc taaagatgcc acataatcta ataaatgaaa tagagattca aacgtggcgt
    15781 catcgtccgt ttcgaccatt tccgaaaaga actcgggcat aaactctatg atttctctgg
    15841 acgtggtgtt gtcgaaactc tcaaagtacg cagtcaggaa cgtgcgcgac atgtcgtcgg
    15901 gaaactcgcg cggaaacatg ttgttgtaac cgaacgggtc ccatagcgcc aaaaccaaat
    15961 ctgccagcgt caatagaatg agcacgatgc cgacaatgga gctggcttgg atagcgattc
    16021 gagttaacta ctgtccgttt ttatttgtgt aataaaatga acggaacgct gaattctgga
//
