LOCUS       sequence-379953-       10278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid encodes S. pyogenes Cas9 and mCherry separated by a T2A 
            peptide under an Aedes aegypti polyubiquitin promoter. It also 
            expresses a sgRNA scaffold under a Culex quinquefasciatus U6 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10278)
  AUTHORS   Torres TZB, Prince BC, Robison A, Ruckert C
  TITLE     Optimized In Vitro CRISPR/Cas9 Gene Editing Tool in the West Nile 
            Virus Mosquito Vector, Culex quinquefasciatus.
  JOURNAL   Insects. 2022 Sep 19;13(9):856. doi: 10.3390/insects13090856.
  PUBMED    36135557
REFERENCE   2  (bases 1 to 10278)
  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: "Insects."; 
            date: "2022-09-19"; volume: "13(9):856. doi"; pages: " 
            10.3390/insects13090856"
COMMENT     Sequence Label: pUb-Cas9-mCherry_cU6:6-sgRNA
FEATURES             Location/Qualifiers
     source          1..10278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(49..637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(129..148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(808..1599,1600..1668))
                     /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     1431..1450
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1669..1773)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3349..3358
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3361..3381
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3406..7506
                     /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             3406..7506
                     /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             7507..7554
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7570..7623
                     /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             7648..8352
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(7790..7808)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8058..8077
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             8356..8409
                     /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             8416..9219
                     /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(8479..8498)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9089..9108
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9227..9361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9251..9270)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     9305..9324
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9362..9378
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9362..9378
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        10042..10117
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 acgcctattt ttataggtta accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
       61 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      121 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      181 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      241 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      301 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      361 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      421 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      481 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      541 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      601 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      661 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      721 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      781 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      841 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
      901 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
      961 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     1021 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     1081 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc
     1141 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     1201 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     1261 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     1321 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     1381 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc
     1441 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1501 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1561 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1621 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1681 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1741 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     1801 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcg
     1861 gctatcttta catgtagctt gtgcattgaa tccaattata atttgccttg gcaccagctg
     1921 agccagacaa gaaagaaagc ttcccagaag tatatcgatt tagaagggtt gacgtcactt
     1981 gctgactgca ctaatacagc aaatgatgca attagaatga ttcaagtgaa attcccaaat
     2041 tactgatttt tctctggatt tggttatcag attacattcg aagctaagat tagctaccga
     2101 aattgtcgat caaatcagga aatcctttct ctatcgaaaa aggcattcgc acatcttcct
     2161 ctctatgcca tatacacgaa gggtaggtac attgacgtct ttgccagaag ttgaactgca
     2221 tcgttcaagg tacagaatga acgactaaca gacacaagca cgttttgctg tccattcaga
     2281 cacagggatg gtacccatat tcgatcgata tagagccatc caaccgaaca gaggtatatg
     2341 tatgaatgta ttgctgaaat tttctagaag tacaaccacc actacgacag tgtctataaa
     2401 acgcccctgc aaaggcgaaa ccagctcaat cgaatacgtt tcctagtgga gtgaacatta
     2461 cgcggcccaa gtaagcagtg ccagtgcaag tgaagtgaag tctctagtga aaaagagtga
     2521 tccaattagc cagaggagaa aatttcagag tgaacaaagc tttattcaaa ggacaattac
     2581 tattaaattg gtgaaagtgc atttcggtga agggaatctt ctagtgaagg taggtaaatt
     2641 aattgatgaa attatagcta tgagcgaaaa ctagtttggt gaatgattcc tttgtctttg
     2701 aatgagcaaa ctattttcca agatggcgac tattgagctt tgagtgatta gtgaaaattt
     2761 gcaacgcagt ttcatcatca ttgataaaac ccaattgtga ttcacagcga taatcatatt
     2821 tcgttgaatc atcgctacta attgaattaa atttctagaa taataagaat aacgtatttg
     2881 ctccgtcaca tatctaaaat aaatattttg atggtaatta cccattaagg taatattaac
     2941 acatatcgag aaaaaccttg aggaaatcgt gaaaacttga agatacgcaa tttcaaaact
     3001 acgtagttca aagtcgaaaa caagttaatt tttcacttaa aagtagggcg ttgttgtgac
     3061 gtcatcacct tcaagtgtat atttttcact tggcctgcga ctgcaaacgc agacaaagca
     3121 aaacaagttt aaaacctgtc gtgtcgtgct cgaagccaaa ggcaatgaat caatatcaaa
     3181 tgagagtttg catttcacaa ccaattactc aagcgtttcc tcgtttcttt ttctgctcaa
     3241 cagagatttc aacaccaagc tcgataagct tgttcgaatc tcgagtgcgc gcttccggag
     3301 gtatacacct aggcggtacc actgcagtga attcggagct ctaccggtgc caccatggcc
     3361 ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc
     3421 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     3481 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     3541 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     3601 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     3661 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     3721 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     3781 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     3841 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     3901 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     3961 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     4021 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     4081 gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg
     4141 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     4201 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     4261 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     4321 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     4381 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     4441 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     4501 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     4561 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     4621 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     4681 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     4741 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     4801 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     4861 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     4921 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     4981 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     5041 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     5101 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     5161 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     5221 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     5281 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     5341 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     5401 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     5461 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     5521 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     5581 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     5641 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     5701 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     5761 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     5821 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     5881 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     5941 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
     6001 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     6061 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     6121 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     6181 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     6241 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     6301 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     6361 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     6421 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     6481 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     6541 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     6601 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     6661 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     6721 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     6781 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     6841 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     6901 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     6961 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     7021 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     7081 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     7141 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     7201 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     7261 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     7321 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     7381 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     7441 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     7501 ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagggatca
     7561 ggcaagcttg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     7621 cctgctagcg gtagcggcag cggtagcgtg agcaagggcg aggaggataa catggccatc
     7681 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     7741 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     7801 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     7861 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     7921 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     7981 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     8041 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     8101 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     8161 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     8221 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     8281 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     8341 gagctgtaca agcttgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat
     8401 cccggccctg ctagcatggg atcggccatt gaacaagatg gattgcacgc aggttctccg
     8461 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     8521 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     8581 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     8641 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     8701 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     8761 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     8821 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     8881 gtcgatcagg atgatctgga cgaagaacat caggggctcg cgccagccga actgttcgcc
     8941 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     9001 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     9061 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagaactt
     9121 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     9181 cgcatcgcct tctatcgcct tcttgacgag ttcttctgac tcgagggatc cagacatgat
     9241 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     9301 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     9361 tcaggaaaca gctatgacca tgattacgcc aagcttggta ccgagctcgg atccactagt
     9421 aacggccgcc agtgtgctgg aattcgccct tcaaaatcta cgaagataat aaggtctact
     9481 attacattta ataaatattc ttatttaatt tgagctgtgt aaaattagtg cgtttatccg
     9541 gggaatgttt atattttatc cgaggatttc ccgaaattga ttaaaaaaat caaatctctt
     9601 aaaaaattag aaacaacctg ttaaaaataa gcttatagaa aaaatgtaaa gtgttgctac
     9661 aaggacaatc aattaatcat cataacgaaa acttttagga tcttgaaaag cacgtcccga
     9721 gatgggaaag tgtgtttcat ttccaggcga tccttttcgc ttcacgaaat ttttgattgc
     9781 tacctcgttt agagccctaa gacaaaatac ttcttcaaaa aaaagcgaac tcgaacccaa
     9841 actttgtact actatgctaa aatgtctgca tgcatgggat gtcgcgcaaa cgagtccaat
     9901 aagaaataaa actgctcgaa ttcaacgtga cgctacccgc aatgctctac ccttaacttt
     9961 atacatcgct agaaccgcgt tcgcattcgc taactatata agcactttcc cggccctaca
    10021 ttcggaagag cgagctcttc cgttttagag ctagaaatag caagttaaaa taaggctagt
    10081 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttagatctg gaaaaatgat
    10141 gtgacagtgg aaatgagaag taggtgcatc tgcaaagatt tgattcagag ttgatgccat
    10201 tcatgatttt tttttagaag tatcatttgt gtttttagaa tatagaatta attcttgaag
    10261 acgaaagggc ctcgtgat
//
