LOCUS       sequence-348671-        8802 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of Drosophila codon optimized spCas9 under Ae. aegypti 
            Polyubiquitin promoter (AAEL003888).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8802)
  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 8802)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pAaePUb::Cas9-2A-Neo
FEATURES             Location/Qualifiers
     source          1..8802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1522..1531
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1531..1596
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1603..1623
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1648..5748
                     /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             1648..5748
                     /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             5749..5796
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5800..5853
                     /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             5860..6663
                     /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(5923..5942)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6533..6552
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6671..6805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6695..6714)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6749..6768
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             join(7001..7069,7070..7861)
                     /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(7219..7238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8032..8620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8521..8540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8774..8791
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggcaact
       61 cgtgaaaggt aggcggatca gcggccgcag catgcgctag ctatctttac atgtagcttg
      121 tgcattgaat ccaattataa tttgccttgg caccagctga gccagacaag aaagaaagct
      181 tcccagaagt atatcgattt agaagggttg acgtcacttg ctgactgcac taatacagca
      241 aatgatgcaa ttagaatgat tcaagtgaaa ttcccaaatt actgattttt ctctggattt
      301 ggttatcaga ttacattcga agctaagatt agctaccgaa attgtcgatc aaatcaggaa
      361 atcctttctc tatcgaaaaa ggcattcgca catcttcctc tctatgccat atacacgaag
      421 ggtaggtaca ttgacgtctt tgccagaagt tgaactgcat cgttcaaggt acagaatgaa
      481 cgactaacag acacaagcac gttttgctgt ccattcagac acagggatgg tacccatatt
      541 cgatcgatat agagccatcc aaccgaacag aggtatatgt atgaatgtat tgctgaaatt
      601 ttctagaagt acaaccacca ctacgacagt gtctataaaa cgcccctgca aaggcgaaac
      661 cagctcaatc gaatacgttt cctagtggag tgaacattac gcggcccaag taagcagtgc
      721 cagtgcaagt gaagtgaagt ctctagtgaa aaagagtgat ccaattagcc agaggagaaa
      781 atttcagagt gaacaaagct ttattcaaag gacaattact attaaattgg tgaaagtgca
      841 tttcggtgaa gggaatcttc tagtgaaggt aggtaaatta attgatgaaa ttatagctat
      901 gagcgaaaac tagtttggtg aatgattcct ttgtctttga atgagcaaac tattttccaa
      961 gatggcgact attgagcttt gagtgattag tgaaaatttg caacgcagtt tcatcatcat
     1021 tgataaaacc caattgtgat tcacagcgat aatcatattt cgttgaatca tcgctactaa
     1081 ttgaattaaa tttctagaat aataagaata acgtatttgc tccgtcacat atctaaaata
     1141 aatattttga tggtaattac ccattaaggt aatattaaca catatcgaga aaaaccttga
     1201 ggaaatcgtg aaaacttgaa gatacgcaat ttcaaaacta cgtagttcaa agtcgaaaac
     1261 aagttaattt ttcacttaaa agtagggcgt tgttgtgacg tcatcacctt caagtgtata
     1321 tttttcactt ggcctgcgac tgcaaacgca gacaaagcaa aacaagttta aaacctgtcg
     1381 tgtcgtgctc gaagccaaag gcaatgaatc aatatcaaat gagagtttgc atttcacaac
     1441 caattactca agcgtttcct cgtttctttt tctgctcaac agagatttca acggatatcc
     1501 agagaccccg gatcggggta cgccaccatg gattacaagg atcacgatgg cgactacaag
     1561 gaccatgaca tcgactataa ggacgacgac gataagatgg cccccaagaa gaagcgcaaa
     1621 gtgggcatcc atggcgtgcc agccgccgat aagaagtaca gcatcggcct ggatatcggc
     1681 accaacagcg tgggctgggc cgtgatcacc gatgagtaca aggtgcccag caagaagttc
     1741 aaggtgctgg gcaacaccga tcgccacagc atcaagaaga acctgatcgg cgccctgctg
     1801 ttcgatagcg gagaaaccgc cgaggccacc cgcctgaagc gcaccgcccg ccgccgctat
     1861 acccgccgca agaatcgcat ctgctatctg caagagatct tcagcaacga gatggccaag
     1921 gtggacgata gcttcttcca ccgcctggag gagtccttcc tggtggagga ggataagaag
     1981 cacgagcgcc accccatctt cggcaacatc gtggatgagg tggcctacca cgagaagtac
     2041 cccaccatct accatctgcg caagaagctg gtggatagca ccgataaggc cgatctgcgc
     2101 ctgatctacc tggccctggc ccacatgatc aagttccgcg gccacttcct gatcgagggc
     2161 gatctgaacc ccgataactc cgatgtggat aagctgttca tccagctggt gcagacctac
     2221 aaccagctgt tcgaggagaa ccccatcaac gccagcggcg tggacgccaa ggccattctg
     2281 agtgcccgcc tgagcaagag tcgccgcctg gagaatctga tcgcccagct gccaggcgag
     2341 aagaagaacg gcctgttcgg caacctgatt gccctgagcc tgggcctgac ccccaacttc
     2401 aagagcaact tcgatctggc cgaggatgcc aagctgcagc tgagcaagga tacctacgat
     2461 gatgatctgg ataacctgct ggcccagatc ggcgatcagt acgccgacct gtttctggcc
     2521 gccaagaacc tgtccgatgc catcctgctg agcgacatcc tgcgcgtgaa caccgagatc
     2581 accaaggccc cactgagcgc ctcgatgatc aagcgctacg atgagcacca ccaggatctg
     2641 accctgctga aggccctcgt gcgccagcag ctgcccgaga agtataagga gatcttcttc
     2701 gaccagagca agaacggcta cgccggctac atcgatggcg gagccagcca ggaggagttc
     2761 tacaagttca tcaagcccat cctggagaag atggatggca ccgaggagct gctcgtgaag
     2821 ctgaaccgcg aggatctgct gcgcaagcag cgcaccttcg ataacggcag catcccccac
     2881 cagatccacc tgggagagct gcacgccatt ctgcgccgcc aggaggattt ttacccgttc
     2941 ctgaaggata accgcgagaa gattgagaag atcctgacct tccggatccc ctactacgtg
     3001 ggaccactgg cccgcggcaa tagccgcttt gcctggatga cccgcaagag cgaggaaacc
     3061 atcaccccct ggaacttcga ggaggtggtg gataagggcg ccagcgccca gagcttcatc
     3121 gagcgcatga ccaacttcga caagaatctg cccaacgaga aggtgctgcc caagcacagc
     3181 ctgctgtacg agtacttcac cgtgtacaac gagctgacga aagtgaaata cgtgaccgag
     3241 ggaatgcgca agcccgcctt cctgagcgga gagcagaaga aggccatcgt ggacctgctg
     3301 tttaagacca accgcaaagt gaccgtgaag cagctgaagg aggactactt caagaagatc
     3361 gagtgcttcg acagcgtgga gatctcgggc gtggaggatc gcttcaatgc cagcctgggc
     3421 acgtaccacg atctgctgaa gattatcaag gacaaggact tcctggacaa cgaggagaat
     3481 gaggacattc tggaggacat cgtgctgacc ctgaccctgt ttgaggatcg cgagatgatc
     3541 gaggagcgcc tcaagaccta cgcccatctg tttgatgaca aagtgatgaa gcagctgaag
     3601 cgccgccgct acaccggatg gggacgcctg agtcgcaagc tgatcaacgg catccgcgat
     3661 aagcagagcg gcaagacgat cctggatttc ctgaagtccg atggcttcgc caatcgcaac
     3721 ttcatgcagc tgatccacga cgattcgctg accttcaagg aggatattca gaaggcccag
     3781 gtgtccggcc agggcgatag cctgcatgag cacattgcca acctggccgg cagccccgcc
     3841 attaagaagg gcattctgca gaccgtgaag gtggtggacg agctcgtgaa agtgatgggc
     3901 cgccacaagc ccgagaatat cgtgattgag atggcccgcg agaaccagac cacccagaag
     3961 ggccagaaga acagccgcga gcgcatgaag cgcatcgagg agggcattaa ggagctgggc
     4021 agccagattc tgaaggagca ccccgtggag aacacccagc tgcagaacga gaagctgtac
     4081 ctgtactacc tgcagaacgg ccgcgatatg tacgtggacc aggagctgga catcaaccgc
     4141 ctgtccgact acgatgtgga ccatatcgtg ccgcagagct ttctgaagga tgacagcatc
     4201 gataacaagg tgctgacccg cagcgataag aaccgcggca agtccgataa cgtgccctcg
     4261 gaggaggtcg tgaagaagat gaagaactat tggcgccagc tgctgaacgc caagctgatt
     4321 acccagcgca agttcgacaa tctgaccaag gccgagcgcg gaggcctgag cgagctggat
     4381 aaggccggct tcatcaagcg ccagctggtg gaaacccgcc agattaccaa gcacgtggcc
     4441 cagattctgg atagccgcat gaacacgaag tacgacgaga atgataagct gattcgcgaa
     4501 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccgcaagga tttccagttc
     4561 tataaagtgc gcgagatcaa caactaccac cacgcccacg atgcctacct gaacgccgtc
     4621 gtgggaaccg ccctgattaa gaagtatccc aagctggaga gcgagttcgt gtacggcgat
     4681 tataaggtgt acgacgtgcg caagatgatc gccaagtccg agcaggagat cggcaaggcc
     4741 accgccaagt actttttcta cagcaacatc atgaatttct tcaagaccga gattaccctg
     4801 gccaacggcg agatccgcaa gcgcccactg atcgaaacga atggcgaaac cggcgagatt
     4861 gtgtgggaca agggccgcga ttttgccacc gtgcgcaagg tgctgtccat gccccaagtg
     4921 aacattgtga aaaagaccga ggtgcagacg ggcggcttca gcaaggagtc cattctgccg
     4981 aagcgcaaca gcgacaagct gatcgcccgc aagaaggatt gggaccccaa gaagtacggc
     5041 ggcttcgata gccccaccgt ggcctatagt gtgctggtgg tggccaaggt ggagaagggc
     5101 aagtccaaga agctgaagtc cgtgaaggag ctgctgggca tcaccatcat ggagcgcagc
     5161 agctttgaga agaacccaat cgatttcctg gaggccaagg gctacaagga agtgaagaag
     5221 gatctgatca tcaagctgcc gaagtactcc ctgttcgagc tggagaacgg acgcaagcgc
     5281 atgctggcca gtgccggcga gctgcagaag ggaaacgagc tggccctgcc ctcgaaatat
     5341 gtgaacttcc tgtacctggc cagccactac gagaagctga agggcagccc cgaggataac
     5401 gagcagaagc agctgtttgt ggagcagcac aagcactacc tggacgagat cattgagcag
     5461 atcagcgagt tcagcaagcg cgtgatcctg gccgacgcca atctggataa ggtgctgagc
     5521 gcctacaaca agcaccgcga caagccaatc cgcgagcagg ccgagaacat catccacctg
     5581 ttcaccctga ccaacctggg agccccagcc gccttcaagt acttcgatac caccatcgat
     5641 cgcaagcgct ataccagcac caaggaggtg ctggacgcca ccctgatcca ccagagcatc
     5701 accggcctgt acgaaacccg catcgatctg tcgcagctgg gaggcgataa gcgcccagcc
     5761 gccaccaaga aggccggaca ggccaaaaag aagaagcctg agggcagagg aagtcttcta
     5821 acatgcggtg acgtggagga gaatcccggc cctgctagca tgggatcggc cattgaacaa
     5881 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5941 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6001 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     6061 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6121 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6181 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6241 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6301 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga acatcagggg
     6361 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     6421 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6481 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6541 acccgtgata ttgctgaaga acttggcggc gaatgggctg accgcttcct cgtgctttac
     6601 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6661 tgactcgagg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     6721 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6781 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     6841 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     6901 tgattatgat cagtcgagcg taattcaaac agggttctgg cgtcgttctc gtactgtttt
     6961 ccccaggcca gtgctttagc gttattgaaa aaggaagagt atgagtattc aacatttccg
     7021 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7081 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     7141 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     7201 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     7261 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     7321 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7381 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     7441 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7501 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7561 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     7621 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7681 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     7741 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     7801 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     7861 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     7921 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7981 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     8041 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     8101 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     8161 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     8221 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     8281 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     8341 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     8401 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     8461 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     8521 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     8581 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     8641 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     8701 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     8761 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa ga
//
