LOCUS       sequence-333381-       16134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cas9 in Culex quinquefasciatus or other mosquitoes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16134)
  AUTHORS   Feng X, Lopez Del Amo V, Mameli E, Lee M, Bishop AL, Perrimon N, 
            Gantz VM
  TITLE     Optimized CRISPR tools and site-directed transgenesis towards gene 
            drive development in Culex quinquefasciatus mosquitoes.
  JOURNAL   Nat Commun. 2021 May 20;12(1):2960. doi: 10.1038/s41467-021-23239-0.
  PUBMED    34017003
REFERENCE   2  (bases 1 to 16134)
  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: "2021-05-20"; volume: "12(1):2960. doi"; pages: " 
            10.1038/s41467-021-23239-0"
COMMENT     Sequence Label: pVMG0173_Cq-Actin5c-Cas9
FEATURES             Location/Qualifiers
     source          1..16134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(69..88)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     279..300
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             754..1548
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(808..827)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1418..1437
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(1566..1634,1635..2426)
                     /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(1784..1803)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2597..3185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3086..3105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3339..3356
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3473..3494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3509..3514,3515..3532,3533..3539)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3547..3563
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3552..3574
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3571..3587
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3571..3587
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(3653..3664)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(4364..4384)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4391..8494)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(4391..8494)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(15933..15952)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(15934..15952)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(15959..15976)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(15959..15975)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15968..15990)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(16116..16134,1..410)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
       61 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      121 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      181 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      241 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      301 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      361 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      421 tcagggcgca agggctgcta aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg
      481 tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca
      541 aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta
      601 tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc
      661 tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga
      721 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
      781 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      841 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
      901 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
      961 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     1021 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatccca ccttgctcct
     1081 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     1141 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     1201 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     1261 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     1321 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     1381 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     1441 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     1501 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat tgaaaaagga
     1561 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1621 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1681 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1741 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1801 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1861 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1921 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1981 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2041 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2101 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2161 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2221 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2281 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2341 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2401 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2461 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     2521 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2581 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2641 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2701 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2761 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2821 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2881 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2941 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3001 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3061 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3121 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3181 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3241 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3301 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3361 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3421 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     3481 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     3541 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     3601 agcttggtac cgagctcgga tccactagta acggccgcca gtgtgctgga attcgccctt
     3661 ctatttgatg aacgacgcac tctcggaata aattaattgc taataaatat tttgacaagc
     3721 ctaaacacaa aacgatacaa aacggtcatt cgatttgggc agttggtttt tttcgcttct
     3781 agcttctttt ttaaactttt ttttcgtgat tttcttattt tttctgactt cgcgctccta
     3841 caaaattaca aaaaagaaaa atgaaagtat cgctaacttg agtaactgtc tgtctttttt
     3901 ttttttggtt ctctcactcg cgctcgctcg cacactcgaa gtttaacgcg ctcgctcgct
     3961 atctcaacat ggaataatat gataattgcg tatgtgtgtg tgtttttgtg tgggtgacta
     4021 aataattcac aaaatcaaat cattaaataa acgtgtggta cttcgatatt ataagtaata
     4081 atacaggaat tgtcttttta tttttaagtt gttctcaaat gtgtgtgttg tttgtgtgta
     4141 tgtgtttttt gagtcattca acacttcgga tgttcttgct gggaatgatt tttgagatga
     4201 tgttgcgctg ttgttgttgc tgttgatttt tcctcgcttt gatggaaagt atctggtttt
     4261 ttgtttgttt ttttaattat aaaggagaag gacgacgatc acgcggccgc gatcggggtc
     4321 ctgtgcaggg gatggtgttg ttgcggcgtg aaggcgctta ttacaccttg cgcttcttct
     4381 tcggcgaacc atcaccaccc agctgcgaca gatcgatgcg cgtctcgtac agacccgtga
     4441 tcgactggtg gatcagcgtg gcatccagca cctccttcgt cgacgtgtag cgcttgcgat
     4501 cgatcgtcgt atcgaagtac ttgaaggcgg ccggggcacc caggttcgtc agcgtgaaca
     4561 ggtggatgat gttctcggcc tgctcgcgga tcggcttatc gcggtgcttg ttgtaggccg
     4621 acagcacctt atccaggttg gcatcggcca ggatcacgcg cttcgagaac tccgagatct
     4681 gctcgatgat ctcatccagg tagtgcttgt gctgctccac gaacagctgc ttctgctcgt
     4741 tatcctccgg cgaacccttc agcttctcgt agtgcgaggc caggtacagg aagttcacgt
     4801 acttcgacgg cagggccagc tcgttaccct tctgcagctc accggccgag gccagcatgc
     4861 gcttgcgacc gttctccagc tcgaacagcg agtacttcgg cagcttgatg atcagatcct
     4921 tcttcacctc cttgtaaccc ttggcctcca ggaaatcgat cgggttcttc tcgaacgacg
     4981 agcgctccat gatcgtgata cccagcagct ccttcaccga cttcagcttc ttcgacttac
     5041 ccttctccac cttggccacc accagcaccg agtaggccac cgtcggcgaa tcgaaaccac
     5101 cgtacttctt cggatcccaa tccttcttgc gggcgatcag cttatccgag ttgcgcttcg
     5161 gcaggatcga ctccttcgag aaaccacccg tctgcacctc cgtcttcttc acgatgttca
     5221 cctgcggcat cgacagcacc ttgcgcaccg tggcgaaatc gcgaccctta tcccacacga
     5281 tctcacccgt ctcaccgttc gtctcgatca gcgggcgctt gcggatctca ccgttggcca
     5341 gcgtgatctc cgtcttgaag aagttcatga tgttcgagta gaagaagtac ttggccgtgg
     5401 ccttaccgat ctcctgctcc gacttggcga tcatcttgcg cacatcgtac accttgtaat
     5461 caccgtacac gaactccgac tccagcttcg ggtacttctt gatcagggcc gtacccacca
     5521 cggcgttcag gtaggcatcg tgggcgtggt ggtagttgtt gatctcgcgc accttgtaga
     5581 actggaaatc cttgcggaaa tccgacacca gcttcgactt cagcgtgatc accttcacct
     5641 cgcggatcag cttatcgttc tcatcgtact tcgtgttcat gcgcgaatcc aggatctggg
     5701 ccacgtgctt cgtgatctgg cgcgtctcca ccagctggcg cttgatgaaa ccggccttat
     5761 ccagctccga cagaccaccg cgctcggcct tcgtcaggtt atcgaacttg cgctgcgtga
     5821 tcagcttggc gttcagcagc tggcgccagt agttcttcat cttcttcacc acctcctccg
     5881 acggcacgtt atccgactta ccgcggttct tatccgagcg cgtcagcacc ttgttatcga
     5941 tcgaatcatc cttcaggaac gactgcggca cgatgtgatc cacatcgtaa tccgacaggc
     6001 ggttgatatc cagctcctga tccacgtaca tatcgcgacc gttctgcagg tagtacaggt
     6061 acagcttctc gttctgcagc tgcgtgttct ccaccgggtg ctccttcagg atctgcgaac
     6121 ccagctcctt gataccctcc tcgatgcgct tcatgcgctc gcgcgagttc ttctgaccct
     6181 tctgcgtcgt ctggttctcg cgggccatct cgatcacgat gttctccggc ttgtggcgac
     6241 ccatcacctt caccagctca tccaccacct tcaccgtctg caggataccc ttcttgatgg
     6301 ccggcgaacc ggccaggttg gcgatgtgct cgtgcagcga atcaccctga cccgacacct
     6361 gggccttctg gatatcctcc ttgaacgtca gcgaatcatc gtggatcagc tgcatgaagt
     6421 tgcggttggc gaaaccatcc gacttcagga aatccaggat cgtcttaccc gactgcttat
     6481 cgcggatacc gttgatcagc ttgcgcgaca ggcgacccca acccgtgtag cggcggcgct
     6541 tcagctgctt catcacctta tcatcgaaca ggtgggcgta cgtcttcagg cgctcctcga
     6601 tcatctcgcg atcctcgaac agcgtcagcg tcagcacgat atcctccagg atatcctcgt
     6661 tctcctcgtt atccaggaaa tccttatcct tgatgatctt cagcagatcg tggtacgtac
     6721 ccagcgaggc gttgaagcga tcctccacac ccgagatctc caccgaatcg aagcactcga
     6781 tcttcttgaa gtaatcctcc ttcagctgct tcaccgtcac cttgcggttc gtcttgaaca
     6841 gcagatccac gatggccttc ttctgctcac ccgacaggaa ggccggcttg cgcataccct
     6901 ccgtcacgta cttcaccttc gtcagctcgt tgtacaccgt gaagtactcg tacagcagcg
     6961 agtgcttcgg cagcaccttc tcgttcggca ggttcttatc gaagttcgtc atgcgctcga
     7021 tgaacgactg ggccgaggca cccttatcca ccacctcctc gaagttccac ggcgtgatcg
     7081 tctcctccga cttgcgcgtc atccaggcga agcgcgagtt accgcgggcc agcggaccca
     7141 cgtagtacgg gatgcggaac gtcaggatct tctcgatctt ctcgcggtta tccttcagga
     7201 acgggtagaa atcctcctgg cggcgcagga tggcgtgcag ctcacccagg tggatctggt
     7261 gcgggatcga accgttatcg aacgtgcgct gcttgcgcag cagatcctcg cggttcagct
     7321 tcaccagcag ctcctccgta ccatccatct tctccaggat cggcttgatg aacttgtaga
     7381 actcctcctg cgaggcacca ccatcgatgt aaccggcgta accgttcttc gactgatcga
     7441 agaagatctc cttgtacttc tccggcagct gctggcgcac cagggccttc agcagcgtca
     7501 gatcctggtg gtgctcatcg tagcgcttga tcatcgaggc cgacagcggg gccttcgtga
     7561 tctccgtgtt cacgcgcagg atatccgaca gcaggatggc atccgacagg ttcttggcgg
     7621 ccaggaacag atcggcgtac tgatcaccga tctgggccag caggttatcc agatcatcat
     7681 cgtacgtatc cttcgacagc tgcagcttgg catcctcggc cagatcgaag ttcgacttga
     7741 agttcggcgt cagacccagc gacagggcga tcaggttacc gaacagaccg ttcttcttct
     7801 cacccggcag ctgggcgatc aggttctcca ggcggcgcga cttcgacagg cgggccgaca
     7861 ggatggcctt ggcatccaca cccgaggcgt tgatcgggtt ctcctcgaac agctggttgt
     7921 acgtctgcac cagctggatg aacagcttat ccacatccga gttatccggg ttcagatcac
     7981 cctcgatcag gaagtgaccg cggaacttga tcatgtgggc cagggccagg tagatcaggc
     8041 gcagatcggc cttatccgtc gaatccacca gcttcttgcg caggtggtag atcgtcgggt
     8101 acttctcgtg gtaggccacc tcatccacga tgttaccgaa gatcgggtgg cgctcgtgct
     8161 tcttatcctc ctccaccagg aacgactcct ccaggcggtg gaagaacgaa tcatccacct
     8221 tggccatctc gttcgagaag atctcctgca ggtagcagat gcggttcttg cggcgcgtgt
     8281 agcggcggcg ggccgtgcgc ttcaggcgcg tggcctcggc cgtctcaccc gaatcgaaca
     8341 gcagggcacc gatcaggttc ttcttgatcg agtggcgatc cgtgttaccc agcaccttga
     8401 acttcttcga cggcaccttg tactcatccg tgatcacggc ccaacccacc gagttcgtac
     8461 cgatatccag accgatcgag tacttcttat ccatgttgat atctgcgcgc aagaagggga
     8521 gaagattcag attagattgc cagattcaac ggtacaaagc ttcgcgagag agagagattg
     8581 caacaaccgt tgggattaac tgttatctac acgcgcgcca cggacaagat aatgcgaggc
     8641 cacgtcagcg gaaagtacca catgaccaac ccaacgcagt taattaaatt acgatcgaat
     8701 tacttgcgtt gaaatctctc cacaaactcg gtatcgtgcg ctctctccgt ctctcagttg
     8761 gttcgtgtct tgcgcgcttt ctcccttcct ccgtctgcgg ggttctgcag gtagcgagga
     8821 ccacgcagac aacgcccacg cagaaaaagg gaccgtcttc ccgggaaggg agaggggaga
     8881 aagagggcaa gacaagcgag ctgaaaattg ttcgactgtg caggcaggga acctgtttct
     8941 acggcggggg tccgtttctt gagtgtttct tgacgcgggt agacgacttt gcgggggatt
     9001 tggaaccggg tagagtcggc aacggcgggg gtgtgttaat tgattttgag gaaagagact
     9061 tgagtgagca attagcggca gacggtgacg ctgggaggta aaaatttgac gatttaggct
     9121 gatttgcacg aaaatgaggt gatgaaatcg gctttgaaaa atcatcaaat ggataaattt
     9181 taatgttttt cctaaaacat tgcagaaata cattgattga aacgttgttc tacgtcaatt
     9241 attgaacatt tacagtcaat ttgatgattc ctgcttgaca acaaatgtta atgaaagatt
     9301 aagtttaaac ttgaaacatt aaaatattct taaattgtca agcgtagtcc cacgtcaatc
     9361 aaataaataa ttaattccct atacaacatg tcaaatttga agcaagtttg acaagttttt
     9421 ttcaaaatcc gggaaatttt tatttaatta aaaaaaaaaa caaaattaaa tctacaaatt
     9481 tcaaaaagtt gataaaatta agcaaaattt taccccactt tcccctttaa aaaatctcac
     9541 caaaacccaa gccgcttgac tcaactgtca agagaggggt gcaacaactt tttttcatct
     9601 ccgttttatt ccaaatttcc cagaagagcg atcaagaacc taggctcttc ttcttttctc
     9661 ttcccctcac acattcacag cagcataatt ggatacaaac gccgcttgct gtgtgagatg
     9721 acgtggggtg ggctgcactt tggtgcgcgc cctcctctgc agttgaatct tccttttcgc
     9781 tctttggctg tgtgagtgag aaaaggatca cactttttct ccttcttttt tttcaagttg
     9841 ggagaagtca acctttttgg gcttgtctag ccacttaaaa tatgtcaaaa tttaaaattt
     9901 ttaaaattta aatttccaac aatcttcaaa aatgttgaag tttaaaaagg atttgccaga
     9961 atcgcaataa aaatcgattc gagccggcat tttggcacgt ccccacgcac acactctcac
    10021 actgcagacc tccggccgga acaaaatgcc ggccaacgcc ccacacaccg gccacaaccc
    10081 ctcttaccct tttcttacaa aagtgagaaa aatgaccgtt acacaggttc tacttcaact
    10141 tcaaaaaaac tacacgaaat tttccagaaa atttaaaaaa acgcaccgtt tgaaaattct
    10201 tttgacagca gctaacaaaa gaaagggaac caccacactt ggattgcttt gatttgagtg
    10261 actttagaat aatttgcacc ggcacacaca cgcgaccgca ccagccgaac gacaacgagc
    10321 gaatgagagg tgggcttggg cttcgggctc ggatggtggc tctccttcgc taacggctct
    10381 tttcgccctc tctcattctc gcgagagcgc gcaactttcg aaaagttctc acccgagccg
    10441 aatctgacgc gagagaaaac gattacgaac ttttcaaaag cttccaccgg atctttctgg
    10501 aaggcacttc gcggcctgga gaatgagaga gcgggagcga gctaacccgc tcgcgctctc
    10561 acccttttac ggagcgtcga ccaatcgcgt gaacatgacc atataaggca atgtttgaaa
    10621 aatgctcaat ttccagcggt ccaagaaggc cggttttcgc acacgtgaac ttgccggtaa
    10681 agatggtggt gggaatggga gaggaagaga gagctttata aggtgtgtgt gtgcgcgaac
    10741 ttgaaaagct tgcgggggtt gcttggagag gggaaggaaa gtcagccgac gcgtcatccg
    10801 gtggggtggt tccacggttg aaattttgtg ataaatttga gtttaaaata ataattttat
    10861 agaaactttt caaaaatttc ttaaaatgtt gcctgtcaaa ggaaacttaa aagagttaga
    10921 tttttttact tgttcaaaca tttttttttg caaattcaat tgaaaaaaaa tttggaagat
    10981 gttcaattga aaagcaatca aaatcaaaat ttttgaagtt tcttgaacag attcttcaac
    11041 aaacatacat ccagcaaaac aaacgtgctt cgaatgtcaa acatcgctcg acacgtagga
    11101 aatgcaaaaa atccaccaat ttatgtaact caaaagaggt ggctagacgt gacaatttcc
    11161 ccatcaacct tcaagaactt gccaaacaaa agaaagttga taaaatttgg gcgcgctgca
    11221 ggtagaaaca cttgcaacca agataaaaac tcgttttgca ggtaaaaata gcaaaatttc
    11281 gaggttcaaa cagcccttgg cgtagtttga agtgtgcggt gcacaaacag ggctggaatt
    11341 gtttgtaaac aaatcgttac atgcacggtg gaatgtcaga gctggctttc aaggtgatga
    11401 cggcggtttg gtgggcgcat caagtaggcc acgtgtgctt atcaggtgga aagtgggtta
    11461 cttttcaatg ttgagtttga tagacttttt gttcttagta aaatttaaga gccagttttg
    11521 acaatagcaa cttgcctata aatggcattt aggatgaaaa taaggcttga gatttattta
    11581 tttttgcgtg ggacaaacat tttgatcaaa atttgtacga gcctaaaaag attttgaaaa
    11641 ttaaattgat tatcgattta atgcactgat gtagacaaat ttatgttgat atttgtcaaa
    11701 attgaatcta aatctgcaag attttgtctg aaatttatgt ttgatttgat attattttta
    11761 gtttaatcat attttcgcca cttttgttga tgaaatcttt aaaaacttcg tgagaattca
    11821 taaaaatgtc catctgatca acattttcca cacaatttcc accactttcc agcgcacatt
    11881 agccacgttg acttagcctg cttctcgcag ccaaaagcgg tacatctccg agttgcattc
    11941 cagcgctgtc tcgccaccaa actttccccg cactgtctct catgattgac gtgggactac
    12001 accccacgtc gttattgttg ccacgtgttt cgagtggaac caccacccaa aaaaatccgg
    12061 acgagctgca ttccacccgg cttttgccgc acccaaaacc aaatcctgcg cgcggttcgt
    12121 cgcttaatcc ggaacccgtt gaagtcactt tagagttcat tgttttttca aatgtctttt
    12181 tgggcacaaa tcactgcatt tttttgcggg gcacaacttg gtaaatgtcc tttgtagtcc
    12241 ttttttgagc acttgtcgaa caattttcac ttgctttttt ccgatgattt gtttttttcc
    12301 atagacgacg aaacttgtta cttacatttg gtttattaaa tacaaaaaag cctaaattac
    12361 aaataaatta attgtgtgct gtgagtttct gcgctggtgt ttcgtagaaa cgtaggctgg
    12421 tagctggtgc tttttcacag aaaataagtt aataaacagc tactactagt ttgttcgatc
    12481 tgttcgatct ctgtctctgg tgtgttcaca gcacgattac aagaactgta gtgatacaat
    12541 ttttaaacaa cactcagatt ataaatagta cccctccaat ccggacgggt atcccaacgg
    12601 ttgcctcggt tgattcgggg ttcgctcgcg cttatgacca tatatggtat tggcagtccg
    12661 ccggaaccac cacagacgcg ttttttctcc agagaagaaa ctcacacact ctcacaaaca
    12721 gagagcgaac gaacgattcg gtgggcgctc gcagctagcg aaccggcata agcaggcggt
    12781 gacgacgacg acggcgagga accaacggaa gtatcaagcg cgatagagag gagaaaagaa
    12841 gcagccgagt aggctatgaa gtgcccaggg ttgggcacac acactcttct tcgttgaccg
    12901 atgcaccatc tcttcatgca tatgcatcgc cagagcagcg tgtgtagaaa agcatatcgg
    12961 cgtgtggggt gcggcgccac gccgcaccca gccctcttgg atcccacttt tgacagtttg
    13021 gggttcaatc aaatactttt ttcttcttaa ttgtcagacg catctaatga ctttatggca
    13081 ccctacttca cttacttgtt tcatgttttg gcgttgccgg ttttccgatc ggagcaagtc
    13141 atgcacaatt tcgaggaatc gcttgaacga tggtggcggc ggttacgaat atccaacagc
    13201 agagccaact ttgataacaa agtgataaga ggtaatcttg ggggtgtcac gtcacctagt
    13261 ttgagttggt tgtttttttt ctttgcagaa ttcttttgaa atagataaca aatggtcgcg
    13321 ttaaaggtgg aaccatcgat ggtattgtgt aagaggtccc ccccccctcg tgttatcagt
    13381 tcgagtggtc tttgttctga attatgtgaa gaatgtcaac gcgggaatga aggtcgtgaa
    13441 ggggattttt ttgcgcttgc tgataaaaag tttgatgaca aatcaattaa gtgtggagtt
    13501 tttttaaggt ccaataaact atttttttac ttaaggcgga gaaaacaaat aagaaaaatt
    13561 tggtttattg gaccttttaa aaaaagactc tgaaagtgtg tttggacttt taaaaatgtt
    13621 agtgttagtt ttttgccaaa tgaattttta atcatatttt tacagttttt aaaaaaatat
    13681 tttattttag aaaaattaaa aaagagtcct tgaaaaactt gtcgttttta ttaaactaat
    13741 ttcaccagta actaaacgta cagacaatca gtctgtgctc aaagtttgcc taagacatca
    13801 aatcgatcag aaaccccgtt ctcaagatcc cgagtttttg aatatttagg ttgcattttt
    13861 gtttggaaag ctggctgttt tgtttataaa ggaagtactc acaatgtttt gaattatgat
    13921 aaaattaggg tacctccaaa aagttgtaac aattttattt ttatgtttta agcatgcaca
    13981 attgtatact tgcggtctgt ttagtctgtg tggaacggac attttaaaat ggtcaaatac
    14041 aggtccaaga ccaaatttgg gataaaaatg tcgttattaa atcatccaaa gtgaaatatt
    14101 agctaggact ttggataatg aaaacgcttt tttcgcttgt tttttttgtt tgcaaagttc
    14161 tgtgtatgta ttaaaattgt attattttaa ctcacaacct tctgctctgg atgcagaggg
    14221 gatgaaaatt aaaaacttgc tgtaaaaata caaacaaatt aaaaaagttc gagtttgaag
    14281 tctaatttaa aactgtgcct cttttttaca ccctgaccaa aaatcctgat tttctaagtt
    14341 taatatctca ctttccatac gatttttttt gagttcagat tctacaacca taaaaaatgg
    14401 ttatatgagt tgaactgaaa aattcgtatg aaaagaaggg aataatgatt tttggtaagg
    14461 gtgtacagag taacctgatt tgacaatttc ggctaaacac aaaaaaaaaa cacttgaaaa
    14521 tgttttatga agagttaaat tgtagtaatt gtattttttt tttgatattt ttttttaagg
    14581 atcgagacaa tacatatttt tttttatttt ttttgtcctt ctgctctggt taagttcaga
    14641 agggatggaa ataaaaaact tactgccaaa agtttacgaa gaaattttaa aataacataa
    14701 aatgtttgac gtctaatttc aatctgtgct ccaaattttt ttagagtgta tcctgttttg
    14761 acaattttga cttaaaacaa caaagtaaaa caagtttaat ggcctatcta caatcacttt
    14821 gcttagaaaa tttcacttag cttaggaatt tgaatcaatg gaaatgaatc tgagtttcta
    14881 caatggaaaa gtaatcaaat tagaaactga cgtttggttt ggaaaatttc aaaatctacg
    14941 gtaagttgac gtttccatat caaaggtaaa caaacaactg catagcaacg tatatcagcc
    15001 cagcaagttt tctaagttga ttgtggatga cgaacgtaag ttgcagactt tcctaagtaa
    15061 ctactttaca tagatgttct aagctaagtg attgtagata ggccataagt ccaatttttt
    15121 gcgttacgta ataaaagaac gctccctttg tggtcaaaag ggggcataga gttatctacg
    15181 cgcgcatgta ttgcgtgtac gtacacgaaa aagattgagg ttaaattttg tatcggccag
    15241 caaaacaaat ggcgtgctag tgtgcgtgag atcgggctta gataactcta ttgagctgat
    15301 ctgaataagc tttatcggtg ttgagatcca gaaaaaagat ccgacagcaa tttgtactga
    15361 tttgacgttt gctgagggtg ccaaaacaat tgcaaaagac aaatattgct gtgaggattt
    15421 gttgattcga atttcgctac ttcgaatatt ggctatttcg aatgctcttc aattcacatt
    15481 tgaatcaaaa agcactcagt tgttaaaatt gagttgtcaa actgatgttt acattttgac
    15541 tcctttgttt gtcgtacttt tgacagttgt tactcgttca gatgagtgaa tctgctctgt
    15601 atcattgtta tctacaaaac ttttcaaaaa acaaagaatt attctaaaat ccccaaaaaa
    15661 atcccccaat gtttacactc ccaagataag cggaccttga catcaagata gcatcttatc
    15721 acccgctctc gagtgaatgt caaaaaggcc tcctccatta agtaaacaaa ccgctgcgga
    15781 gcgggggaaa aatatttaca caaattactc tcacagagtc caccgcatcc cagccaagcc
    15841 gggggtaaat tggcaaattg gccaagggcg aattctgcag atatccatca cactggcggc
    15901 cgctcgagca tgcatctaga gggcccaatt cgccctatag tgagtcgtat tacaattcac
    15961 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
    16021 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
    16081 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgc
//
