LOCUS       sequence-371153-       14854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9-VPR under the Ae. aegypti polyubiquitin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14854)
  AUTHORS   Bui M, Dalla Benetta E, Dong Y, Zhao Y, Yang T, Li M, Antoshechkin 
            IA, Buchman A, Bottino-Rojas V, James AA, Perry MW, Dimopoulos G, 
            Akbari OS
  TITLE     CRISPR mediated transactivation in the human disease vector Aedes 
            aegypti.
  JOURNAL   PLoS Pathog. 2023 Jan 19;19(1):e1010842. doi: 
            10.1371/journal.ppat.1010842. eCollection 2023 Jan.
  PUBMED    36656895
REFERENCE   2  (bases 1 to 14854)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog."; date: "2023-01-19"; volume: "19(1):e1010842. doi"; pages: 
            " 10.1371/journal.ppat.1010842. eCollection 2023 Jan"
COMMENT     Sequence Label: OA-986F (AAEL003877-dCas9-VPR)
FEATURES             Location/Qualifiers
     source          1..14854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(142..730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(222..241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(901..1692,1693..1761))
                     /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     1524..1543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1762..1866)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1934..1952
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1990..2012)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2112..2131
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2325..2347
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2339..2356
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2340..2356
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   3060..3122
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4156..4225
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     regulatory      5823..5832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5829..5855,5856..5858,5859..8342,8343..8348,8349..8414,
                     8415..8417,8418..9932)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9945..9965
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9990..10139
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             10164..10184
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10614..10970
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             10989..11558
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             join(12796..12798,12799..12801,12802..13476)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(12979..12999)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     13197..13216
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     13391..13414
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    13590..13711
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13627..13646)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13681..13700
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   14379..14413
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(14594..14610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14594..14610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14607..14629)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14618..14634
                     /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)."
     promoter        complement(join(14642..14648,14649..14666,14667..14672))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14687..14708
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14825..14842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
       61 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      121 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      181 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      241 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      301 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      361 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      421 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      481 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      541 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
      601 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
      661 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
      721 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
      781 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
      841 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
      901 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
      961 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1021 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     1081 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1141 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1201 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1261 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1321 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1381 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1441 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1501 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1561 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1621 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1681 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     1741 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     1801 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     1861 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     1921 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     1981 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     2041 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     2101 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     2161 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     2221 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     2281 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     2341 taaaacgacg gccagtgcca agctttgttt aaaaatataa caaaattgtg atcccacaaa
     2401 atgaagtggg gcaaaatcaa ataattaata gtgtccgtaa acttgttggt cttcaacttt
     2461 ttgaggaaca cgttggacgg caaatccgtg actataacac aagttgattt aataatttta
     2521 gccaacacgt cgggctgcgt gttttttgcc gacgcgtctg tgtacacgtt gattaactgg
     2581 tcgattaaac tgttgaaata atttaatttt tggttcttct ttaaatctgt gatgaaattt
     2641 tttaaaataa ctttaaattc ttcattggta aaaaatgcca cgttttgcaa cttgtgaggg
     2701 tctaatatga ggtcaaactc agtaggagtt ttatccaaaa aagaaaacat gattacgtct
     2761 gtacacgaac gcgtattaac gcagagtgca aagtataaga gggttaaaaa atatatttta
     2821 cgcaccatat acgcatcggg ttgatatcgt taatatggat caatttgaac agttgattaa
     2881 cgtgtctctg ctcaagtctt tgatcaaaac gcaaatcgac gaaaatgtgt cggacaatat
     2941 caagtcgatg agcgaaaaac taaaaaggct agaatacgac aatctcacag acagcgttga
     3001 gatatacggt attcacgaca gcaggctgaa taataaaaaa attagaaact attatttaac
     3061 cctagaaaga taatcatatt gtgacgtacg ttaaagataa tcatgcgtaa aattgacgca
     3121 tgtgttttat cggtctgtat atcgaggttt atttattaat ttgaatagat attaagtttt
     3181 attatattta cacttacata ctaataataa attcaacaaa caatttattt atgtttattt
     3241 atttattaaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa acttttaaac
     3301 attctctctt ttacaaaaat aaacttattt tgtactttaa aaacagtcat gttgtattat
     3361 aaaataagta attagcttaa cttatacata atagaaacaa attatactta ttagtcagtc
     3421 agaaacaact ttggcacata tcaatattat gctctcgaca aataactttt ttgcattttt
     3481 tgcacgatgc atttgccttt cgccttattt tagaggggca gtaagtacag taagtacgtt
     3541 ttttcattac tggctcttca gtactgtcat ctgatgtacc aggcacttca tttggcaaaa
     3601 tattagagat attatcgcgc aaatatctct tcaaagtagg agcttctaaa cgcttacgca
     3661 taaacgatga cgtcaggctc atgtaaaggt ttctcataaa ttttttgcga ctttgaacct
     3721 tttctccctt gctactgaca ttatggctgt atataataaa agaatttatg caggcaatgt
     3781 ttatcattcc gtacaataat gccataggcc acctattcgt cttcctactg caggtcatca
     3841 cagaacacat ttggtctagc gtgtccactc cgcctttagt ttgattataa tacataacca
     3901 tttgcggttt accggtactt tcgttgatag aagcatcctc atcacaagat gataataagt
     3961 ataccatctt agctggcttc ggtttatatg agacgagagt aaggggtccg tcaaaacaaa
     4021 acatcgatgt tcccactggc ctggagcgac tgtttttcag tacttccggt atctcgcgtt
     4081 tgtttgatcg cacggttccc acaatggtta attcgagctc gcccggggtc ctaggtcgac
     4141 gatgtaggtc acggtctcga agccgcggtg cgggtgccag ggcgtgccct tgggctcccc
     4201 gggcgcgtac tccacctcac ccatctggtc catcatgatg aacgggtcga ggtggcggta
     4261 gttgatcccg gcgaacgcgc ggcgcaccgg gaagccctcg ccctcgaaac cgctgggcgc
     4321 ggtggtcacg gtgagcacgg gacgtgcgac ggcgtcggcg ggtgcggata cgcggggcag
     4381 cgtcagcggg ttctcgacgg tcacggcggg catgtcgacg cggccgctat ctttacatgt
     4441 agcttgtgca ttgaatccaa ttataatttg ccttggcacc agctgagcca gacaagaaag
     4501 aaagcttccc agaagtatat cgatttagaa gggttgacgt cactttgctg actgcactaa
     4561 tacagcaaat gatacaatta gaatgattca agtgaaattc ccaaattact gctttgtctc
     4621 tggatttggt tatcagatta cattcgaagc taagaatagc taccgaaatt gtcgatcaaa
     4681 tcaggaaatc ctttctctat cgaaaaaggc attcgcacat ctttctctgt atgccatata
     4741 cacgaatggt aggtacattg acgtctttgc cagaagttga actgcatcgt tcaaggtaca
     4801 gaatgaacga ctaacagaca caagcacgtt ttgctgtcca ttcagacaca gggatggtac
     4861 ccatagtcga tcgatttaga gccatccaac cgaacagagg tatatgtatg aatggattgc
     4921 agaaattttc tagaagtaca accaccacta cggcagtgtc tataaaacgc ccctgcaaag
     4981 gcaaaaccag ctcaatcgaa tacgtttcct agtggagtga acattacgcg gtccaagtaa
     5041 gcagtgccag tgcaagtgaa gtgaagtctc tagtgaaaaa gagtgatcca attagccaga
     5101 ggagaaaatt tcagagtgaa caaagctttg ttcaaaggac aattactatt aaatttgtga
     5161 aagtgcattt cggtgaaggg aatcttctag tgaaggtagg taaattaaat gatgaaatta
     5221 tagctatgag cgaaaactag tttggtgaat gattcctttg tctttgaatg agcaaactat
     5281 tttccaagat ggcgactatt gagctttgag tgattagtga aaatttgcaa cgcagtttca
     5341 tcatcattga taaaacccaa ttgtgattca cggcgataat catatttcgt tgaatcatcg
     5401 ctgctaattg aattaaattt ctagagcaag cgcgaattcg ccatatttct aaaattaaat
     5461 attgtggtga taattaccca ttaaggtaat attaacacat atcgagaaaa accttgagga
     5521 aatcgtgaaa acttgaagat acgcaatttc caaactacgt agttcaaagt cgaaaacaag
     5581 ttaatttttc actaaaaagt agggcgttgt tgtgacgtca tcaccttcaa gtgtatattt
     5641 ttcacttggc ctgcgactgc aaacgcagac aaagcaaaac aagtttaaaa cctgtcgtgt
     5701 cgtgctcgaa gccaaaggca atgaatcaat atcaaatgag agtttgcatt tcacaaccaa
     5761 ttactgaagc gtttcctcgt ttctttttct gctcaacaga gatttcaacg aatgtttaaa
     5821 ctgccaccat ggacaagaag tactccattg ggctcgctat cggcacaaac agcgtcggct
     5881 gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa attcaaagtt ctgggcaata
     5941 ccgatcgcca cagcataaag aagaacctca ttggcgccct cctgttcgac tccggggaga
     6001 cggccgaagc cacgcggctc aaaagaacag cacggcgcag atatacccgc agaaagaatc
     6061 ggatctgcta cctgcaggag atctttagta atgagatggc taaggtggat gactctttct
     6121 tccataggct ggaggagtcc tttttggtgg aggaggataa aaagcacgag cgccacccaa
     6181 tctttggcaa tatcgtggac gaggtggcgt accatgaaaa gtacccaacc atatatcatc
     6241 tgaggaagaa gcttgtagac agtactgata aggctgactt gcggttgatc tatctcgcgc
     6301 tggcgcatat gatcaaattt cggggacact tcctcatcga gggggacctg aacccagaca
     6361 acagcgatgt cgataaactc tttatccaac tggttcagac ttacaatcag cttttcgaag
     6421 agaacccgat caacgcatcc ggagttgacg ccaaagcaat cctgagcgct aggctgtcca
     6481 aatcccggcg gctcgaaaac ctcatcgcac agctccctgg ggagaagaag aacggcctgt
     6541 ttggtaatct tatcgccctg tcactcgggc tgacccccaa ctttaaatct aacttcgacc
     6601 tggccgaaga tgccaagctt caactgagca aagacaccta cgatgatgat ctcgacaatc
     6661 tgctggccca gatcggcgac cagtacgcag accttttttt ggcggcaaag aacctgtcag
     6721 acgccattct gctgagtgat attctgcgag tgaacacgga gatcaccaaa gctccgctga
     6781 gcgctagtat gatcaagcgc tatgatgagc accaccaaga cttgactttg ctgaaggccc
     6841 ttgtcagaca gcaactgcct gagaagtaca aggaaatttt cttcgatcag tctaaaaatg
     6901 gctacgccgg atacattgac ggcggagcaa gccaggagga attttacaaa tttattaagc
     6961 ccatcttgga aaaaatggac ggcaccgagg agctgctggt aaagcttaac agagaagatc
     7021 tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc ccaccagatt cacctgggcg
     7081 aactgcacgc tatcctcagg cggcaagagg atttctaccc ctttttgaaa gataacaggg
     7141 aaaagattga gaaaatcctc acatttcgga taccctacta tgtaggcccc ctcgcccggg
     7201 gaaattccag attcgcgtgg atgactcgca aatcagaaga gaccatcact ccctggaact
     7261 tcgaggaagt cgtggataag ggggcctctg cccagtcctt catcgaaagg atgactaact
     7321 ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg tacgagtact
     7381 tcacagttta taacgagctc accaaggtca aatacgtcac agaagggatg agaaagccag
     7441 cattcctgtc tggagagcag aagaaagcta tcgtggacct cctcttcaag acgaaccgga
     7501 aagttaccgt gaaacagctc aaagaagact atttcaaaaa gattgaatgt ttcgactctg
     7561 ttgaaatcag cggagtggag gatcgcttca acgcatccct gggaacgtat cacgatctcc
     7621 tgaaaatcat taaagacaag gacttcctgg acaatgagga gaacgaggac attcttgagg
     7681 acattgtcct cacccttacg ttgtttgaag atagggagat gattgaagaa cgcttgaaaa
     7741 cttacgctca tctcttcgac gacaaagtca tgaaacagct caagaggcgc cgatatacag
     7801 gatgggggcg gctgtcaaga aaactgatca atgggatccg agacaagcag agtggaaaga
     7861 caatcctgga ttttcttaag tccgatggat ttgccaaccg gaacttcatg cagttgatcc
     7921 atgatgactc tctcaccttt aaggaggaca tccagaaagc acaagtttct ggccaggggg
     7981 acagtcttca cgagcacatc gctaatcttg caggtagccc agctatcaaa aagggaatac
     8041 tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat gggaaggcat aagcccgaga
     8101 atatcgttat cgagatggcc cgagagaacc aaactaccca gaagggacag aagaacagta
     8161 gggaaaggat gaagaggatt gaagagggta taaaagaact ggggtcccaa atccttaagg
     8221 aacacccagt tgaaaacacc cagcttcaga atgagaagct ctacctgtac tacctgcaga
     8281 acggcaggga catgtacgtg gatcaggaac tggacatcaa tcggctctcc gactacgacg
     8341 tggctgctat cgtgccccag tcttttctca aagatgattc tattgataat aaagtgttga
     8401 caagatccga taaagctaga gggaagagtg ataacgtccc ctcagaagaa gttgtcaaga
     8461 aaatgaaaaa ttattggcgg cagctgctga acgccaaact gatcacacaa cggaagttcg
     8521 ataatctgac taaggctgaa cgaggtggcc tgtctgagtt ggataaagcc ggcttcatca
     8581 aaaggcagct tgttgagaca cgccagatca ccaagcacgt ggcccaaatt ctcgattcac
     8641 gcatgaacac caagtacgat gaaaatgaca aactgattcg agaggtgaaa gttattactc
     8701 tgaagtctaa gctggtctca gatttcagaa aggactttca gttttataag gtgagagaga
     8761 tcaacaatta ccaccatgcg catgatgcct acctgaatgc agtggtaggc actgcactta
     8821 tcaaaaaata tcccaagctt gaatctgaat ttgtttacgg agactataaa gtgtacgatg
     8881 ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa ggccaccgct aagtacttct
     8941 tttacagcaa tattatgaat tttttcaaga ccgagattac actggccaat ggagagattc
     9001 ggaagcgacc acttatcgaa acaaacggag aaacaggaga aatcgtgtgg gacaagggta
     9061 gggatttcgc gacagtccgg aaggtcctgt ccatgccgca ggtgaacatc gttaaaaaga
     9121 ccgaagtaca gaccggaggc ttctccaagg aaagtatcct cccgaaaagg aacagcgaca
     9181 agctgatcgc acgcaaaaaa gattgggacc ccaagaaata cggcggattc gattctccta
     9241 cagtcgctta cagtgtactg gttgtggcca aagtggagaa agggaagtct aaaaaactca
     9301 aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg atcaagcttc gaaaaaaacc
     9361 ccatcgactt tctcgaggcg aaaggatata aagaggtcaa aaaagacctc atcattaagc
     9421 ttcccaagta ctctctcttt gagcttgaaa acggccggaa acgaatgctc gctagtgcgg
     9481 gcgagctgca gaaaggtaac gagctggcac tgccctctaa atacgttaat ttcttgtatc
     9541 tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga taatgagcag aagcagctgt
     9601 tcgtggaaca acacaaacac taccttgatg agatcatcga gcaaataagc gaattctcca
     9661 aaagagtgat cctcgccgac gctaacctcg ataaggtgct ttctgcttac aataagcaca
     9721 gggataagcc catcagggag caggcagaaa acattatcca cttgtttact ctgaccaact
     9781 tgggcgcgcc tgcagccttc aagtacttcg acaccaccat agacagaaag cggtacacct
     9841 ctacaaagga ggtcctggac gccacactga ttcatcagtc aattacgggg ctctatgaaa
     9901 caagaatcga cctctctcag ctcggtggag acagcagggc tgaccccaag aagaagagga
     9961 aggtggaggc cagcggttcc ggacgggctg acgcattgga cgattttgat ctggatatgc
    10021 tgggaagtga cgccctcgat gattttgacc ttgacatgct tggttcggat gcccttgatg
    10081 actttgacct cgacatgctc ggcagtgacg cccttgatga tttcgacctg gacatgctga
    10141 ttaactctag aagttccgga tctccgaaaa agaaacgcaa agttggtagc cagtacctgc
    10201 ccgacaccga cgaccggcac cggatcgagg aaaagcggaa gcggacctac gagacattca
    10261 agagcatcat gaagaagtcc cccttcagcg gccccaccga ccctagacct ccacctagaa
    10321 gaatcgccgt gcccagcaga tccagcgcca gcgtgccaaa acctgccccc cagccttacc
    10381 ccttcaccag cagcctgagc accatcaact acgacgagtt ccctaccatg gtgttcccca
    10441 gcggccagat ctctcaggcc tctgctctgg ctccagcccc tcctcaggtg ctgcctcagg
    10501 ctcctgctcc tgcaccagct ccagccatgg tgtctgcact ggctcaggca ccagcacccg
    10561 tgcctgtgct ggctcctgga cctccacagg ctgtggctcc accagcccct aaacctacac
    10621 aggccggcga gggcacactg tctgaagctc tgctgcagct gcagttcgac gacgaggatc
    10681 tgggagccct gctgggaaac agcaccgatc ctgccgtgtt caccgacctg gccagcgtgg
    10741 acaacagcga gttccagcag ctgctgaacc agggcatccc tgtggcccct cacaccaccg
    10801 agcccatgct gatggaatac cccgaggcca tcacccggct cgtgacaggc gctcagaggc
    10861 ctcctgatcc agctcctgcc cctctgggag caccaggcct gcctaatgga ctgctgtctg
    10921 gcgacgagga cttcagctct atcgccgaca tggacttctc cgcactgctg ggtagcggat
    10981 cgggatctcg ggattccagg gaagggatgt ttttgccgaa gcctgaggcc ggctccgcta
    11041 ttagtgacgt gtttgagggc cgcgaggtgt gccagccaaa acgaatccgg ccatttcatc
    11101 ctccaggaag tccatgggcc aaccgcccac tccccgccag cctcgcacca acaccaaccg
    11161 gtccagtaca tgagccagtc gggtcactga ccccggcacc agtccctcag ccactggatc
    11221 cagcgcccgc agtgactccc gaggccagtc acctgttgga ggatcccgat gaagagacga
    11281 gccaggctgt caaagccctt cgggagatgg ccgatactgt gattccccag aaggaagagg
    11341 ctgcaatctg tggccaaatg gacctttccc atccgccccc aaggggccat ctggatgagc
    11401 tgacaaccac acttgagtcc atgaccgagg atctgaacct ggactcaccc ctgaccccgg
    11461 aattgaacga gattctggat accttcctga acgacgagtg cctcttgcat gccatgcata
    11521 tcagcacagg actgtccatc ttcgacacat ctctgttttg attaattaac tagaatgaat
    11581 cgtttttaaa ataacaaatc aattgtttta taatattcgt acgattcttt gattatgtaa
    11641 taaaatgtga tcattaggaa gattacgaaa aatataaaaa atatgagttc tgtgtgtata
    11701 acaaatgctg taaacgccac aattgtgttt gttgcaaata aacccatgat tatttgatta
    11761 aaattgttgt tttctttgtt catagacaat agtgtgtttt gcctaaacgt gtactgcata
    11821 aactccatgc gagtgtatag cgagctagtg gctaacgctt gccccaccaa agtagattcg
    11881 tcaaaatcct caatttcatc accctcctcc aagtttaaca tttggccgtc ggaattaact
    11941 tctaaagatg ccacataatc taataaatga aatagagatt caaacgtggc gtcatcgtcc
    12001 gtttcgacca tttccgaaaa gaactcgggc ataaactcta tgatttctct ggacgtggtg
    12061 ttgtcgaaac tctcaaagta cgcagtcagg aacgtgcgcg acatgtcgtc gggaaactcg
    12121 cgcggaaaca tgttgttgta accgaacggg tcccatagcg ccaaaaccaa atctgccagc
    12181 gtcaatagaa tgagcacgat gccgacaatg gagctggctt ggatagcgat tcgagttaac
    12241 ggccggggcg cgcccatact cggtggcctc cccaccacca acttttttgc actgcaaaaa
    12301 aacacgcttt tgcacgcggg cccatacata gtacaaactc tacgtttcgt agactatttt
    12361 acataaatag tctacaccgt tgtatacgct ccaaatacac taccacacat tgaacctttt
    12421 tgcagtgcaa aaaagtacgt gtcggcagtc acgtaggccg gccttatcgg gtcgcgtcct
    12481 gtcacgtacg aatcacatta tcggaccgga cgagtgttgt cttatcgtga caggacgcca
    12541 gcttcctgtg ttgctaaccg cagccggacg caactcctta tcggaacagg acgcgcctcc
    12601 atatcagccg cgcgttatct catgcgcgtg accggacacg aggcgcccgt cccgcttatc
    12661 gcgcctataa atacagcccg caacgatctg gtaaacacag ttgaacagca tctgttacag
    12721 cgacacaaca tgagccggtc caacaacgcc aacgcgccca cgccatccaa ccgccgccgc
    12781 aacctgtctc tggtgatggt gcgctcctcc aagaacgtca tcaaggagtt catgcgcttc
    12841 aaggtgcgca tggagggcac cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
    12901 ggccgcccct acgagggcca caacaccgtg aagctgaagg tgaccaaggg cggccccctg
    12961 cccttcgcct gggacatcct gtccccccag ttccagtacg gctccaaggt gtacgtgaag
    13021 caccccgccg acatccccga ctacaagaag ctgtccttcc ccgagggctt caagtgggag
    13081 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
    13141 gacggctgct tcatctacaa ggtgaagttc atcggcgtga acttcccctc cgacggcccc
    13201 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac
    13261 ggcgtgctga agggcgagat ccacaaggcc ctgaagctga aggacggcgg ccactacctg
    13321 gtggagttca agtccatcta catggccaag aagcccgtgc agctgcccgg ctactactac
    13381 gtggactcca agctggacat cacctcccac aacgaggact acaccatcgt ggagcagtac
    13441 gagcgcaccg agggccgcca ccacctgttc ctgtaggact ctagatcata atcagccata
    13501 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
    13561 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
    13621 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
    13681 gtggtttgtc caaactcatc aatgtatctt aaagcttcta ggccggccga tctcggatct
    13741 gacaatgttc agtgcagaga ctcggctacg cctcgtggac tttgaagttg accaacaatg
    13801 tttattctta cctctaatag tcctctgtgg caaggtcaag attctgttag aagccaatga
    13861 agaacctggt tgttcaataa cattttgttc gtctaatatt tcactaccgc ttgacgttgg
    13921 ctgcacttca tgtacctcat ctataaacgc ttcttctgta tcgctctgga cgtcatcttc
    13981 acttacgtga tctgatattt cactgtcaga atcctcacca acaagctcgt catcgctttg
    14041 cagaagagca gagaggatat gctcatcgtc taaagaacta cccattttat tatatattag
    14101 tcacgatatc tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga
    14161 aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc
    14221 gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa
    14281 gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt
    14341 cgcgctattt agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac
    14401 tatctttcta gggttaaaaa agatttgcgc tttactcgac ctaaacttta aacacgtcat
    14461 agaatcttcg tttgacaaaa accacattgt ggccaagctg tgtgacgcga cgcgcgctaa
    14521 agaatggcaa accaagtcgc gcgagcgtcg actctagagg atccccgggt accgagctcg
    14581 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
    14641 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
    14701 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
    14761 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
    14821 cttcctcgct cactgactcg ctgcgctcgg tcgt
//
