LOCUS       sequence-342956-       11141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eSpCas9 expression vector for Anopheles.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11141)
  AUTHORS   Klug D, Arnold K, Mela-Lopez R, Marois E, Blandin SA
  TITLE     A toolbox of engineered mosquito lines to study salivary gland 
            biology and malaria transmission.
  JOURNAL   PLoS Pathog. 2022 Oct 12;18(10):e1010881. doi: 
            10.1371/journal.ppat.1010881. eCollection 2022 Oct.
  PUBMED    36223382
REFERENCE   2  (bases 1 to 11141)
  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: "PLoS 
            Pathog."; date: "2022-10-12"; volume: "18(10):e1010881. doi"; pages:
            " 10.1371/journal.ppat.1010881. eCollection 2022 Oct"
COMMENT     Sequence Label: pDSARN-vasa-eSpCas9sv40
FEATURES             Location/Qualifiers
     source          1..11141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     256..278
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     270..287
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     271..287
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(337..461)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2771..2836
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2843..2863
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2888..5425,5426..5428,5429..5890,5891..5893,5894..6061,
                     6062..6064,6065..6988)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6989..7036
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    7153..7235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    7396..7478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    7536..7605
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     regulatory      8120..8129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8126..8800
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(8306..8326)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     8718..8741
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             8810..8830
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8971..9105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    9153..9186
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(9253..9352)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(9382..9401)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9383..9401)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9406..9422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9406..9422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9524..9543)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             9535..10344
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(9602..9621)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      10491..11079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10980..10999
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc
      301 ccctacaggt cactaatacc atctaagtag ttgattcata gtgactggat atgttgtgtt
      361 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
      421 tcattttacg tttctcgttc aacttttcta tacaaagttg gtaccggatc ccgatgtaga
      481 acgcgagcaa attcttttcc ttccatgaca gcagcagcta cagtgggaag ccgaacgtca
      541 gacgtgtttg acatgccgaa ctgggcggga aaattacagc gtgcgctttg ttttcaagca
      601 aatcacaact cgctgcaaac aaaaccgttg agaaattgat tgttttataa tttgtattgt
      661 attttatttg ttataataaa ctaaaaagac atactttttg catattttat acataaaaac
      721 atacatgcag cattataaaa cacatataaa ccctccctgt agagtcccgt atcgaaatct
      781 tccatcctag ttgcacagta cgacggacga gtaggccgtg tccgtgcaaa ttccagcttt
      841 tagcagtctt ttgctcggag cactcgcggc gagtcggagg tttctgctga ggtgcttagc
      901 gctaaattag ccaattgctt ttgcaagtga aataaccagc cgaatagtac ttcaaaactc
      961 aggtaagtga actagtttta tagaacaaat gtttgtttgt tagaagttag tgaagtgttt
     1021 gtgaaaaaaa tctctcattt cggcaaaact aacgtaactg atttcaaatt gaattattgt
     1081 tttgtgatgt tatattattt catccagttg attagtattt tcttagttat gttcaaaata
     1141 cagttaaatt aaatttcatt tcatttactc ataaaataat ctcttggctt atttaatttt
     1201 tctcgaattc gcttgtattg ttcagtagca cgcgccattc gccctttgtt tcattttgta
     1261 cctgctccca ctaacacact ggcagtgcga aacaaaagcc ttcgcacgcg ttgctggtat
     1321 tagagtgtgt gcgtgtgtgt gttgagcgct ctgtcaaaat cggctgttgc cgccggtacc
     1381 gaaattgcct gttcgcacgc tgttcgtaaa cattccgtgg tgtgtatcgt gtgttgtgca
     1441 tgttgcgcgc ctcccccctt ttgatagcag gctgccgtgg ctgccgtggt gtgtggcgca
     1501 gttgagtttt tggattaatt ttctaaggaa atggcacgag aagagcggtg gcagtgtgtt
     1561 ggtttgctct gtcccttcct ttctgtgtga agtgttctta cagcacagca cgtatccacc
     1621 accgcacaca gagcaggcaa ggaagtggaa gtgaacaagt gtgctgcgca tgcatgtgtg
     1681 tggggggcat tttagctgag atcgtcgtta tttgagaagc ggtatagggg ccagtcggtg
     1741 tcgacgtacg gaagcggttt agttttaatc caagcgtatc ccgtcgtgga gtggttgtgt
     1801 ggctctgtgt gctctcatat cagttccaga gtgaggttag tagaatcaca gtccttggcc
     1861 tttttcgtta caagatatcc agaaggatgg cgttatttcc acagcttacc atggtgctct
     1921 tgtttgctcg aatcagggga gaaaaacagt ttcgtgtttc atgaaccgca gttggcactg
     1981 gagcggattc aaaagtcttc gatatgcaat agataagaga gtcgttgggg catagttggg
     2041 aagcctttcc gagatgtgga gtttccgaga ggagaaatgg tgctttcgtg cacgttccgg
     2101 gacagcgggc cccgcgaaga gcatctcgtt gtcgttcatc cggcaataat tgatgcgaaa
     2161 agcgcgcgcg ccactggctt agcgcagtgt acacagtgat attcacctac acacacagag
     2221 gcacacgcct tcacacgcgc gcgtgcttca aaggctactt cggtggcggt gtgtgaggtc
     2281 gcttgcaatg gacaatgaaa atttcgctgg aaaataccat cgtctcttta ggttgcaatg
     2341 ggtgcgggta gagcggtggt cgtcgatatt ggtggtgtag tgtgtgtgtg tgtgtgtgtg
     2401 tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg
     2461 tgtgtgtgtg tgtgtgcaac ggcaattatt ttttgtaata tttcgaccat ctttctttct
     2521 ctctctccac gtgctgctgc tgttgctgct gctgctgcat tgcatgttcc actattcctc
     2581 tcggtttgtg cctgcggacg ccattgctag tcgaaagaga gtcgccgtta gtcgcgcttc
     2641 gagcaacgga cacgtttttt ggttgaaacc aacagctttt ttcatcttcg ggagacacac
     2701 agatctcgaa tcgtacattc ccataaggag aattgtcatc ttccggtgaa taaagaaagg
     2761 aaacaaaatg gactataagg accacgacgg agactacaag gatcatgata ttgattacaa
     2821 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
     2881 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
     2941 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     3001 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
     3061 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     3121 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     3181 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     3241 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     3301 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     3361 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     3421 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     3481 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     3541 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     3601 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     3661 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     3721 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     3781 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     3841 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     3901 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     3961 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     4021 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     4081 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     4141 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     4201 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     4261 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     4321 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     4381 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     4441 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     4501 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     4561 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     4621 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     4681 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     4741 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     4801 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     4861 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     4921 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     4981 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     5041 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     5101 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     5161 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     5221 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     5281 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     5341 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     5401 ccatatcgtg cctcagagct ttctggccga cgactccatc gacaacaagg tgctgaccag
     5461 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     5521 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     5581 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     5641 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     5701 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     5761 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     5821 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     5881 aaagtaccct gcgctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     5941 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     6001 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     6061 ggcgcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     6121 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     6181 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     6241 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     6301 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     6361 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     6421 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     6481 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     6541 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     6601 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     6661 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     6721 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     6781 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     6841 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     6901 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     6961 gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca
     7021 ggcaaaaaag aaaaagtaag aattctagac ataatcagcc ataccacatt tgtagaggtt
     7081 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     7141 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7201 acaaatttca caaataaagc atttttcttc actgcattct agttgtggtt tgtccaaact
     7261 catcaatgta tctaggatgt gcggcttcta gacataatca gccataccac atttgtagag
     7321 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     7381 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     7441 atcacaaatt tcacaaataa agcatttttc ttcactgcat tctagttgtg gtttgtccaa
     7501 actcatcaat gtatctcgac gatgtaggtc acagtctcga agccgcggtg cgggtgccag
     7561 ggcgtgccct tgggctcccc gggcgcgtac tccacctcac ccatctggtc catcatgatg
     7621 aacgggtcga ggtggcggta gttgatcccg gcgaacgcgc ggcgcaccgg gaagccctcg
     7681 ccctcgaaac cgctgggcgc ggtggtcacg gtgagcacgg gacgtgcgac ggcgtcggcg
     7741 ggtgcggata cgcggggcag cgtcagcggg ttctcgacgg tcacggcggg caattcctgc
     7801 agacttccgg tatctcgcgt ttgtttgatc gcacggttcc cacaatggtt aattcgagct
     7861 cgcccgggga tctaattcaa ttagagacta attcaattag agctaattca attaggatcc
     7921 aagcttatcg atttcgaacc ctcgaccgcc ggagtataaa tagaggcgct tcgtctacgg
     7981 agcgacaatt caattcaaac aagcaaagtg aacacgtcgc taagcgaaag ctaagcaaat
     8041 aaacaagcgc agctgaacaa gctaaacaat cggggtaccg ctagagtcga cggtaccgcg
     8101 ggcccgggat ccaccggtcg ccaccatggc ctcctccgag aacgtcatca ccgagttcat
     8161 gcgcttcaag gtgcgcatgg agggcaccgt gaacggccac gagttcgaga tcgagggcga
     8221 gggcgagggc cgcccctacg agggccacaa caccgtgaag ctgaaggtga ccaagggcgg
     8281 ccccctgccc ttcgcctggg acatcctgtc cccccagttc cagtacggct ccaaggtgta
     8341 cgtgaagcac cccgccgaca tccccgacta caagaagctg tccttccccg agggcttcaa
     8401 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggcg accgtgaccc aggactcctc
     8461 cctgcaggac ggctgcttca tctacaaggt gaagttcatc ggcgtgaact tcccctccga
     8521 cggccccgtg atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc
     8581 ccgcgacggc gtgctgaagg gcgagaccca caaggccctg aagctgaagg acggcggcca
     8641 ctacctggtg gagttcaagt ccatctacat ggccaagaag cccgtgcagc tgcccggcta
     8701 ctactacgtg gacgccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     8761 gcagtacgag cgcaccgagg gccgccacca cctgttcctg agatctcgac ccaagaaaaa
     8821 gcggaaggtg gaggacccgt aagatccacc ggatctagat aactgatcat aatcagccat
     8881 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     8941 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     9001 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     9061 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcccgtt tgacggtatc
     9121 gataagcttg atggggatcc ggaaccctta attaccgttc gtataatgta tgctatacga
     9181 agttattagg tccctcgacc tgcagcccgt acggcgcgcc tagagcggcc gccaccgcgg
     9241 tggagctcga gtacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat
     9301 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     9361 ctgcagggcg gccgcgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     9421 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     9481 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     9541 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     9601 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     9661 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     9721 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     9781 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     9841 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     9901 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     9961 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
    10021 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
    10081 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
    10141 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
    10201 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
    10261 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
    10321 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
    10381 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
    10441 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
    10501 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
    10561 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
    10621 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
    10681 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
    10741 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
    10801 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
    10861 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
    10921 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
    10981 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
    11041 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
    11101 ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
