LOCUS       sequence-348641-       10283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Drosophila codon optimized spCas9 under Drosophila 
            Act5C promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10283)
  AUTHORS   Viswanatha R, Mameli E, Rodiger J, Merckaert P, Feitosa-Suntheimer 
            F, Colpitts TM, Mohr SE, Hu Y, Perrimon N
  TITLE     Bioinformatic and cell-based tools for pooled CRISPR knockout 
            screening in mosquitos.
  JOURNAL   Nat Commun. 2021 Nov 24;12(1):6825. doi: 10.1038/s41467-021-27129-3.
  PUBMED    34819517
REFERENCE   2  (bases 1 to 10283)
  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: "Nat 
            Commun."; date: "2021-11-24"; volume: "12(1):6825. doi"; pages: " 
            10.1038/s41467-021-27129-3"
COMMENT     Sequence Label: pDmAct5C::Cas9-2A-Neo
FEATURES             Location/Qualifiers
     source          1..10283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..33
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        97..2606
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     regulatory      2622..2631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2631..2696
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2703..2723
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2748..6848
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2748..6848
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6849..6896
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6900..6953
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6960..7763
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7023..7042)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7633..7652
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7771..7905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7795..7814)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7849..7868
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(8506..9094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8586..8605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9265..10056,10057..10125))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9888..9907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10126..10230)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttaattctat
       61 attctaaaaa cacaaatgat acttctaaaa aaaaatcatg aatggcatca actctgaatc
      121 aaatctttgc agatgcacct acttctcatt tccactgtca catcattttt ccagatctcg
      181 ctgcctgtta tgtggcccac aaaccaagac acgttttatg gccattaaag ctggctgatc
      241 gtcgccaaac accaaataca taatgaatat gtacacattc gagaaagaag cgatcaaaga
      301 agcgtcttcg ggcggagtag gagaatgcgg aggagaagga gaacgagctg atctagtatc
      361 tctccacaat ccaatgccaa ctgaccaact ggccatattc ggagcaattt gaagccaatt
      421 tccatcgcct ggcgatcgct ccattcttgg ctatatgttt ttcaccgtta cccggggcca
      481 ttttcaaaga ctcgtcggca agataagatt gtgtcactcg ctgtctctct tcatttgtcg
      541 aagaatgctg aggaatttcg cgatgacgtc ggcgagtatt ttgaagaatg agaataattt
      601 gtatttatac gaaaatcagt tagtggaatt ttctacaaaa acatgttatc tatagataat
      661 tttgttgcaa aatatgttga ctatgacaaa gattgtatgt atataccttt aatgtattct
      721 cattttctta tgtatttata atggcaatga tgatactgat gatattttaa gatgatgcca
      781 gaccaaaagg cttgaatttc tgcgtctttt gccgaacgca gtgcatgtgc aattgttgtt
      841 ttttggaata ttcaattttc ggactgtccg ctttgatttc agtttcttgg cttattcaaa
      901 aagcaaagta aagccaaaaa agcgagatgg caataccaaa tgcggcaaaa cggtagtgga
      961 aggaaagggg tgcggggcag cggaaggaag ggtggggcgg ggcgtggcgg ggtctgtggc
     1021 tgggcgcgac gtcaccgacg ttggagccac tcctttgacc atgtgtgcgt gtgtgtatta
     1081 ttcgtgtctc gccactcgcc ggttgttttt ttctttttat gctgcgctct ctctagcgcc
     1141 atctcgctta cgcatgctca acgcaccgca tgttgccgtt tccttttatg cgtcattttg
     1201 gctcgaaata ggcaattatt taaacaaaga ttagtcaacg aaaacgctaa aataaataag
     1261 tctacaatat ggttacttat tgccatgtgt gtgcagccaa cgatagcaac aaaagcaaca
     1321 acacaggtgg ctttccctct ttcacttttt gtttgcaagc cgcgtgcgag caagacggca
     1381 cgaccggcaa acgcaattac gctgacaaag agcagacgaa gttttggcga aaaacatcaa
     1441 ggcgcctgat acgaatgcat ttgcaataac aattgcgata tttaatattg tttatgaagc
     1501 tgtttgactt caaaacacac aaaaaaaaaa ataaaacaaa ttatttgaaa gagaattagg
     1561 aatcggacgc ttatcgttag ggtaacaaca agaaatgctt actgagtcac agcctctgga
     1621 aaactgccgc aagccagaga gagagagaaa aagagggaga gcagcttaga ccgcatgtgc
     1681 ttgtgtgtga ggcgtctctc tcttcgtctc tgttgcgcaa acgcatagac tgcactgaaa
     1741 aaatcgatta cctatttttt atgaatgaat atttgcacta ttactattca aaactattaa
     1801 gatagcaatc acattcaata gccaaatact ataccacctg agcgatgcaa cgaaatgatc
     1861 aatttgagca aaaatgctgc atatttagga cggcatcatt atagaaatgc ttcttgctgt
     1921 gtacttttct ctcgtctggc agctgtttcg ccgttattgt taaaaccggc ttaagttagg
     1981 tgtgttttct acgactagtg aatgccctac tagaagatgt gtgttgcaca aaatgtccct
     2041 ggaataacca atttgaagtg cagatagcag taaacgtaag ctaatatgaa tattatttaa
     2101 ctgtaatgtt ttaatatcgc tggacattac taataaaccc actataaaca catgtacata
     2161 tgtatgtttt ggcatacaat gagtagttgg ggaaaaaatg tgtaaaagca ccgtgaccat
     2221 cacagcataa agataaccag ctgaagtatc gaatatgagt aacccccaaa ttgaatcaca
     2281 tgccgcaact gataggaccc atggaagtac actcttcatg gcgatataca agacacacac
     2341 aagcacgaac acccagttgc ggaggaaatt ctccgtaaat gaaaacccaa tcggcgaaca
     2401 attcataccc atatatggta aaagttttga acgcgacttg agagcggaga gcattgcggc
     2461 tgataaggtt ttagcgctaa gcgggcttta taaaacgggc tgcgggacca gttttcatat
     2521 cactaccgtt tgagttcttg tgctgtgtgg atactcctcc cgacacaaag ccgctccatc
     2581 agccagcagt cgtctaatcc agagaccccg gatcggggta cgccaccatg gattacaagg
     2641 atcacgatgg cgactacaag gaccatgaca tcgactataa ggacgacgac gataagatgg
     2701 cccccaagaa gaagcgcaaa gtgggcatcc atggcgtgcc agccgccgat aagaagtaca
     2761 gcatcggcct ggatatcggc accaacagcg tgggctgggc cgtgatcacc gatgagtaca
     2821 aggtgcccag caagaagttc aaggtgctgg gcaacaccga tcgccacagc atcaagaaga
     2881 acctgatcgg cgccctgctg ttcgatagcg gagaaaccgc cgaggccacc cgcctgaagc
     2941 gcaccgcccg ccgccgctat acccgccgca agaatcgcat ctgctatctg caagagatct
     3001 tcagcaacga gatggccaag gtggacgata gcttcttcca ccgcctggag gagtccttcc
     3061 tggtggagga ggataagaag cacgagcgcc accccatctt cggcaacatc gtggatgagg
     3121 tggcctacca cgagaagtac cccaccatct accatctgcg caagaagctg gtggatagca
     3181 ccgataaggc cgatctgcgc ctgatctacc tggccctggc ccacatgatc aagttccgcg
     3241 gccacttcct gatcgagggc gatctgaacc ccgataactc cgatgtggat aagctgttca
     3301 tccagctggt gcagacctac aaccagctgt tcgaggagaa ccccatcaac gccagcggcg
     3361 tggacgccaa ggccattctg agtgcccgcc tgagcaagag tcgccgcctg gagaatctga
     3421 tcgcccagct gccaggcgag aagaagaacg gcctgttcgg caacctgatt gccctgagcc
     3481 tgggcctgac ccccaacttc aagagcaact tcgatctggc cgaggatgcc aagctgcagc
     3541 tgagcaagga tacctacgat gatgatctgg ataacctgct ggcccagatc ggcgatcagt
     3601 acgccgacct gtttctggcc gccaagaacc tgtccgatgc catcctgctg agcgacatcc
     3661 tgcgcgtgaa caccgagatc accaaggccc cactgagcgc ctcgatgatc aagcgctacg
     3721 atgagcacca ccaggatctg accctgctga aggccctcgt gcgccagcag ctgcccgaga
     3781 agtataagga gatcttcttc gaccagagca agaacggcta cgccggctac atcgatggcg
     3841 gagccagcca ggaggagttc tacaagttca tcaagcccat cctggagaag atggatggca
     3901 ccgaggagct gctcgtgaag ctgaaccgcg aggatctgct gcgcaagcag cgcaccttcg
     3961 ataacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcgccgcc
     4021 aggaggattt ttacccgttc ctgaaggata accgcgagaa gattgagaag atcctgacct
     4081 tccggatccc ctactacgtg ggaccactgg cccgcggcaa tagccgcttt gcctggatga
     4141 cccgcaagag cgaggaaacc atcaccccct ggaacttcga ggaggtggtg gataagggcg
     4201 ccagcgccca gagcttcatc gagcgcatga ccaacttcga caagaatctg cccaacgaga
     4261 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtacaac gagctgacga
     4321 aagtgaaata cgtgaccgag ggaatgcgca agcccgcctt cctgagcgga gagcagaaga
     4381 aggccatcgt ggacctgctg tttaagacca accgcaaagt gaccgtgaag cagctgaagg
     4441 aggactactt caagaagatc gagtgcttcg acagcgtgga gatctcgggc gtggaggatc
     4501 gcttcaatgc cagcctgggc acgtaccacg atctgctgaa gattatcaag gacaaggact
     4561 tcctggacaa cgaggagaat gaggacattc tggaggacat cgtgctgacc ctgaccctgt
     4621 ttgaggatcg cgagatgatc gaggagcgcc tcaagaccta cgcccatctg tttgatgaca
     4681 aagtgatgaa gcagctgaag cgccgccgct acaccggatg gggacgcctg agtcgcaagc
     4741 tgatcaacgg catccgcgat aagcagagcg gcaagacgat cctggatttc ctgaagtccg
     4801 atggcttcgc caatcgcaac ttcatgcagc tgatccacga cgattcgctg accttcaagg
     4861 aggatattca gaaggcccag gtgtccggcc agggcgatag cctgcatgag cacattgcca
     4921 acctggccgg cagccccgcc attaagaagg gcattctgca gaccgtgaag gtggtggacg
     4981 agctcgtgaa agtgatgggc cgccacaagc ccgagaatat cgtgattgag atggcccgcg
     5041 agaaccagac cacccagaag ggccagaaga acagccgcga gcgcatgaag cgcatcgagg
     5101 agggcattaa ggagctgggc agccagattc tgaaggagca ccccgtggag aacacccagc
     5161 tgcagaacga gaagctgtac ctgtactacc tgcagaacgg ccgcgatatg tacgtggacc
     5221 aggagctgga catcaaccgc ctgtccgact acgatgtgga ccatatcgtg ccgcagagct
     5281 ttctgaagga tgacagcatc gataacaagg tgctgacccg cagcgataag aaccgcggca
     5341 agtccgataa cgtgccctcg gaggaggtcg tgaagaagat gaagaactat tggcgccagc
     5401 tgctgaacgc caagctgatt acccagcgca agttcgacaa tctgaccaag gccgagcgcg
     5461 gaggcctgag cgagctggat aaggccggct tcatcaagcg ccagctggtg gaaacccgcc
     5521 agattaccaa gcacgtggcc cagattctgg atagccgcat gaacacgaag tacgacgaga
     5581 atgataagct gattcgcgaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     5641 tccgcaagga tttccagttc tataaagtgc gcgagatcaa caactaccac cacgcccacg
     5701 atgcctacct gaacgccgtc gtgggaaccg ccctgattaa gaagtatccc aagctggaga
     5761 gcgagttcgt gtacggcgat tataaggtgt acgacgtgcg caagatgatc gccaagtccg
     5821 agcaggagat cggcaaggcc accgccaagt actttttcta cagcaacatc atgaatttct
     5881 tcaagaccga gattaccctg gccaacggcg agatccgcaa gcgcccactg atcgaaacga
     5941 atggcgaaac cggcgagatt gtgtgggaca agggccgcga ttttgccacc gtgcgcaagg
     6001 tgctgtccat gccccaagtg aacattgtga aaaagaccga ggtgcagacg ggcggcttca
     6061 gcaaggagtc cattctgccg aagcgcaaca gcgacaagct gatcgcccgc aagaaggatt
     6121 gggaccccaa gaagtacggc ggcttcgata gccccaccgt ggcctatagt gtgctggtgg
     6181 tggccaaggt ggagaagggc aagtccaaga agctgaagtc cgtgaaggag ctgctgggca
     6241 tcaccatcat ggagcgcagc agctttgaga agaacccaat cgatttcctg gaggccaagg
     6301 gctacaagga agtgaagaag gatctgatca tcaagctgcc gaagtactcc ctgttcgagc
     6361 tggagaacgg acgcaagcgc atgctggcca gtgccggcga gctgcagaag ggaaacgagc
     6421 tggccctgcc ctcgaaatat gtgaacttcc tgtacctggc cagccactac gagaagctga
     6481 agggcagccc cgaggataac gagcagaagc agctgtttgt ggagcagcac aagcactacc
     6541 tggacgagat cattgagcag atcagcgagt tcagcaagcg cgtgatcctg gccgacgcca
     6601 atctggataa ggtgctgagc gcctacaaca agcaccgcga caagccaatc cgcgagcagg
     6661 ccgagaacat catccacctg ttcaccctga ccaacctggg agccccagcc gccttcaagt
     6721 acttcgatac caccatcgat cgcaagcgct ataccagcac caaggaggtg ctggacgcca
     6781 ccctgatcca ccagagcatc accggcctgt acgaaacccg catcgatctg tcgcagctgg
     6841 gaggcgataa gcgcccagcc gccaccaaga aggccggaca ggccaaaaag aagaagcctg
     6901 agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc cctgctagca
     6961 tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     7021 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     7081 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     7141 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     7201 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     7261 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     7321 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     7381 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     7441 tggacgaaga acatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     7501 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     7561 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     7621 atcaggacat agcgttggct acccgtgata ttgctgaaga acttggcggc gaatgggctg
     7681 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     7741 gccttcttga cgagttcttc tgactcgagg gatccagaca tgataagata cattgatgag
     7801 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     7861 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     7921 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     7981 ctctacaaat gtggtatggc tgattatgat cagtcgaccg atgcccttga gagccttcaa
     8041 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     8101 cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga
     8161 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt
     8221 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca
     8281 ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg cgttcgcgac
     8341 gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca tcgggatgcc
     8401 cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac agcttcaagg
     8461 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     8521 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     8581 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     8641 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     8701 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     8761 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     8821 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     8881 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     8941 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     9001 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     9061 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     9121 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     9181 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     9241 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     9301 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     9361 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     9421 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     9481 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     9541 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
     9601 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     9661 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     9721 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     9781 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     9841 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     9901 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     9961 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
    10021 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
    10081 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
    10141 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    10201 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
    10261 ctaagaaacc attattatca tga
//
