LOCUS       sequence-381998-       10589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GFP-Cas9-NLS and gRNA in A. castellanii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10589)
  AUTHORS   Bisio H, Legendre M, Giry C, Philippe N, Alempic JM, Jeudy S, 
            Abergel C
  TITLE     Evolution of giant pandoravirus revealed by CRISPR/Cas9.
  JOURNAL   Nat Commun. 2023 Jan 26;14(1):428. doi: 10.1038/s41467-023-36145-4.
  PUBMED    36702819
REFERENCE   2  (bases 1 to 10589)
  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: "2023-01-26"; volume: "14(1):428. doi"; pages: " 
            10.1038/s41467-023-36145-4"
COMMENT     Sequence Label: vHB8
FEATURES             Location/Qualifiers
     source          1..10589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(581..583,584..586,587..1297)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNDHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(887..906)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1234..1255
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1304..5404
                     /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             1304..5404
                     /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             5420..5440
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5441..5467
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_RNA        6141..6216
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      6423..6878
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6510..6529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6720..6741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6995..7355
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6995..7186
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7018..7038)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             7586..8380
                     /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(7640..7659)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8250..8269
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8444..8492
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(8533..8552)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8652..8674
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8712..8730)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8798..8902
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8903..8971,8972..9763)
                     /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(9121..9140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9934..10522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10423..10442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcga ggggagatgg ttgaagaagc tgtggaagag gactgggcgg tactaccagc
       61 agcgaacatg ccatcgcccc ccgaggcccc atgtcgcgtc agcagggacg gcgtagggcc
      121 aggatgtctc gtaggggagg gaggcggatt gttcgccccc tgagttgcag atgcggatcg
      181 tgtgctgctc atttgcttca ttctatgcaa ttggatccaa ttggaatttg tagattaatc
      241 tgttggcaag gaaccaaggt cgattgggcg ggaccatcga agagaaacct cagggaaaag
      301 gcatttcaga tttcttttat atttcgcacg gggcgatcaa acagcgagaa aaccgacaga
      361 cacgtgactc tggagttcct catgaaattt ctgatgttta tgaattttga ttggctattc
      421 tgataaagag gcaagccgga gtttatcttg gaatattctc aggccacgtc atactccggc
      481 aggggggcac gggggacaaa agcgggaggg cgaagcggcc gtcttttttc tttgcttccg
      541 aggagagcgt agactatacc aacaccacca atccacccat atggtgagca agggcgagga
      601 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acgaccacaa
      661 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      721 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      781 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      841 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      901 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      961 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1021 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1081 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1141 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1201 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1261 cgccgggatc actctcggca tggacgagct gtacaagctc gaggacaaga agtactccat
     1321 cggtctcgac atcggcacca actcggtcgg ctgggccgtc atcaccgacg agtacaaggt
     1381 gccctccaag aagttcaagg tgctcggcaa cacggacagg cactcgatca agaagaacct
     1441 catcggcgcg ctcctcttcg actcgggaga gacggccgag gccacccgtc tcaagcgcac
     1501 ggctcgtagg cgctacacca ggcgcaagaa ccgcatctgc tacctccagg agatcttctc
     1561 gaacgagatg gccaaggtgg acgactcctt cttccacagg ctggaggaga gcttcctggt
     1621 ggaggaggac aagaagcacg agcgccaccc gatcttcggt aacatcgtgg acgaggtcgc
     1681 ctaccacgag aagtacccca ccatctacca tctccggaag aagctggtcg actcgaccga
     1741 caaggcggac ctgaggctga tctacctggc cctcgcgcac atgatcaagt tcaggggcca
     1801 cttcctcatc gagggtgacc tcaaccccga caactctgac gtggataagc tcttcatcca
     1861 gctcgtccag acctacaacc agctcttcga ggagaacccc atcaacgcgt cgggcgtcga
     1921 cgccaaggcc atcctctccg ctcgcctctc caagtcccgc aggctggaga accttatcgc
     1981 ccagctgccc ggcgagaaga agaacggcct cttcggcaac ctgatcgccc tctctctggg
     2041 cctgaccccg aacttcaaga gcaacttcga cctcgctgag gacgccaagc tgcagctcag
     2101 caaggacacc tacgatgacg acctcgataa cctcctcgct cagatcggcg accagtacgc
     2161 cgaccttttc ctcgccgcca agaacctctc ggacgccatc ctgctctcgg atatcctccg
     2221 cgtcaacacc gagatcacca aggcccccct ctccgcctcc atgatcaagc gctacgacga
     2281 gcaccaccag gacttgacgc tcctgaaggc cctcgtgcgc cagcagctcc ccgagaagta
     2341 caaggagatc ttcttcgatc agtccaagaa cggctacgcc ggctacatcg atggcggtgc
     2401 ctcgcaggag gagttctaca agttcatcaa gcccatcctc gagaagatgg acggcaccga
     2461 ggagctgctc gtgaagctca accgagagga tctcctccgc aagcagcgca ccttcgacaa
     2521 cggcagcatc ccgcaccaga tccacctcgg tgagctccat gccatcctgc gccgccagga
     2581 ggatttctac cccttcctca aggacaaccg cgagaagatc gaaaagatcc ttaccttccg
     2641 catcccctac tacgtgggcc cgctggcccg aggcaactcg cgcttcgcct ggatgacccg
     2701 caagtcggag gagacgatca cgccctggaa cttcgaggag gtggtggaca agggtgcctc
     2761 ggcccagtcg ttcatcgaga ggatgaccaa cttcgacaag aacctcccca acgagaaggt
     2821 cctgcccaag cacagcctgc tctacgagta cttcaccgtg tacaacgagc tcaccaaggt
     2881 caagtacgtc accgagggca tgcgcaagcc cgcgttcctc agcggcgagc agaagaaggc
     2941 gatcgtcgat cttctcttca agaccaaccg caaggtgacg gtcaagcagc tcaaggagga
     3001 ctacttcaag aagatcgagt gcttcgattc tgtggagatc agcggcgttg aggaccgctt
     3061 caacgcctcg ctcggcacct accacgacct gctgaagatc atcaaggaca aggatttcct
     3121 cgacaacgag gagaacgagg acatcctcga ggacatcgtc ctcaccctca ccctcttcga
     3181 ggaccgcgag atgatcgagg agcgcctcaa gacctacgcc cacctcttcg acgataaggt
     3241 gatgaagcag ctgaagcggc ggcgatacac gggctggggc aggctctcga ggaagctgat
     3301 caacggcatc cgcgacaagc agagcggcaa gaccatcctc gacttcctca agtccgacgg
     3361 cttcgcgaac cgcaacttca tgcagctcat ccacgacgac tccctcacgt tcaaggagga
     3421 catccagaag gcccaggtct ccggccaggg cgacagcctc cacgagcaca tcgccaacct
     3481 cgccggcagc cccgccatca agaagggcat tctgcagacc gtgaaggtgg tcgacgaact
     3541 ggtcaaggtc atgggccgcc acaagccgga gaacatcgtg atcgagatgg cgcgcgagaa
     3601 ccagaccacc cagaagggtc agaagaactc ccgtgagcgc atgaagcgta ttgaggaggg
     3661 catcaaggag ctcggttccc agatcctcaa ggagcacccc gtcgagaaca cccagctgca
     3721 gaatgagaag ctgtacctct actacctcca gaacggccgc gacatgtacg tcgaccagga
     3781 gctcgacatc aaccgtcttt cggactacga cgtcgaccac atcgtccccc agtcgttcct
     3841 gaaggacgac tcgatcgaca acaaggtcct gacccgctcc gacaagaacc gcggtaagag
     3901 cgacaacgtc ccctcggagg aggtcgtcaa gaagatgaag aactactgga ggcagctcct
     3961 caacgccaag ctgatcactc agaggaagtt cgacaacctg actaaggccg agcgcggcgg
     4021 ccttagcgag ctggacaagg ccggtttcat caagcgccag ctcgtcgaga cccggcagat
     4081 caccaagcac gtcgcccaga ttctcgattc gaggatgaac acgaagtacg atgagaacga
     4141 caagctcatc cgtgaggtca aggtcatcac cctgaagtcg aagctcgtgt ccgacttccg
     4201 caaggacttc cagttctaca aggtccgcga gatcaacaac taccaccacg cccacgatgc
     4261 ctacctcaac gctgtggtgg gtaccgccct catcaagaag tatcctaagc tcgagtcgga
     4321 gttcgtctac ggcgactaca aggtgtacga cgtgcgcaag atgatcgcta agagcgagca
     4381 ggagatcggt aaggccaccg ccaagtactt cttctactcc aacatcatga acttcttcaa
     4441 gaccgagatc accctggcga acggcgagat ccgcaagcgc cctctcatcg agaccaacgg
     4501 cgagactggc gagatcgtgt gggataaggg cagggacttc gccaccgtga ggaaggtcct
     4561 ctcgatgccg caggtcaaca tcgtcaagaa gacggaggtc caaaccggtg gcttctccaa
     4621 ggagtccatc ctgcccaaga ggaacagcga caagctgatc gcccgcaaga aggactggga
     4681 ccccaagaag tatggcggct tcgacagccc taccgtcgcc tactcggtgc tggtcgtggc
     4741 caaggtcgag aagggcaagt ccaagaagct gaagtccgtc aaggagctcc tcggcatcac
     4801 catcatggag cgctcgtcgt tcgagaagaa cccgatcgac ttcctcgagg cgaagggtta
     4861 caaggaggtg aagaaggacc tcatcattaa gctcccgaag tacagcctct tcgagctgga
     4921 gaacggccgc aagcgcatgc tcgcgtcggc cggtgagctc cagaagggca acgagctcgc
     4981 cctgcccagc aagtacgtga acttcctcta cctggcgtcg cactacgaga agctcaaggg
     5041 ctcgcccgag gacaacgagc agaagcagct gttcgtggag cagcacaagc actacctgga
     5101 cgagatcatc gagcagatct ccgagttcag caagcgcgtg atcctcgccg acgccaacct
     5161 cgacaaggtg ctgtccgcct acaacaagca ccgcgacaag cccatccgcg agcaggccga
     5221 gaacatcatc cacctcttca cgctcaccaa cctgggcgcc ccggccgcct tcaagtactt
     5281 tgacacgacc atcgaccgca agcgctacac ctccaccaag gaggtgctcg acgccactct
     5341 catccaccag agcattaccg gcctctacga gaccaggatc gatctcagcc agctgggcgg
     5401 tgacggagga tccggaccac caaagaagaa gcgtaaggtc tacccatacg acgtcccaga
     5461 ctacgcctaa tctagatgcc cccacctcgt ccgcccaacc caagctctct gcaagtctac
     5521 gctctaccat cctccgaaag catgtctctt gtgctctcga cacgagtcgc gctccaagct
     5581 ctgacctcgc cctgacgaac gcggctcgtt gacaattttg tttacaagac ccaaaaaaaa
     5641 cataaaaaac tctcaaaaaa tgttctcgcc ttcatcaata ttggccatta gccatattat
     5701 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     5761 atcataatat gtacaagtaa agcggcctcg agaattcacg cgtccggagg cctctcccct
     5821 aggtgcagcg ttgaagcccc cgagcccgac gacccaccaa atatcttcgt caccgcatgt
     5881 ggcaagctct ccatcttctt cttcttattc gctttatctt tctttttccg tttctgattt
     5941 cccattcatt tcttttttct attttaggaa aggtccgctc ctttttcgtc ccacaagcga
     6001 aggagacgac cggccaggag tggttcaaga aggcgccagc catgcacttc atgagacaag
     6061 gccggttagc ttagttggta gagcgtggtg ctaataacgc caaggtcgtg ggttcgatcc
     6121 ccatactggc cagcggccgc gttttagagc tagaaatagc aagttaaaat aaggctagtc
     6181 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgactagag tcgacccaac
     6241 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     6301 agcaagtaaa acctctacaa atgtggtaaa atccgataag gatcgatccg ggctggcgta
     6361 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     6421 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6481 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6541 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6601 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     6661 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6721 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     6781 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6841 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     6901 acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa
     6961 taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga
     7021 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     7081 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     7141 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     7201 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     7261 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     7321 gaggcttttt tggaggatca aacgacgcct tgcaacaagc tgagaaaaaa ccaggatcgg
     7381 caaggaagga ttttcaacgg aaacgttggc atcaccgggt ataaaaaggg gccaattttt
     7441 ttgttgattt gttgcgcgaa ttcttgcttt cggcatcgaa ttcaagggag aaggagtcga
     7501 ttcacacata caacaagatc ctaggctttt gcaaaaagct tgattcttct gacacaacag
     7561 tctcgaactt aaggctagag ccaccatgat tgaacaagat ggattgcacg caggttctcc
     7621 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     7681 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     7741 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     7801 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     7861 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     7921 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     7981 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     8041 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     8101 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     8161 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     8221 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     8281 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     8341 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     8401 atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt
     8461 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgcgtatg
     8521 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     8581 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     8641 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     8701 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     8761 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     8821 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     8881 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     8941 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     9001 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     9061 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     9121 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     9181 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     9241 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     9301 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     9361 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     9421 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     9481 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     9541 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     9601 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     9661 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     9721 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     9781 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     9841 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     9901 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     9961 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
    10021 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
    10081 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
    10141 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
    10201 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
    10261 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
    10321 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
    10381 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
    10441 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
    10501 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
    10561 ttgctggcct tttgctcaca tggctcgac
//
