LOCUS       sequence-381993-       10776 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 10776)
  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 10776)
  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: vHB16
FEATURES             Location/Qualifiers
     source          1..10776
                     /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        6154..6229
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        6328..6403
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      6610..7065
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6697..6716)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6907..6928
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7182..7542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7182..7373
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7205..7225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             7773..8567
                     /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(7827..7846)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8437..8456
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8631..8679
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(8720..8739)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8839..8861
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8899..8917)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8985..9089
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9090..9158,9159..9950)
                     /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(9308..9327)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10121..10709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10610..10629
                     /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 cagtgacggc caccaccgaa ccggttttag agctagaaat agcaagttaa
     6181 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgcg acaaggccgg
     6241 ttagcttagt tggtagagcg tggtgctaat aacgccaagg tcgtgggttc gatccccata
     6301 ctggccagat tgtgcgccgc atgtccggtt ttagagctag aaatagcaag ttaaaataag
     6361 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg actagagtcg
     6421 acccaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     6481 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc cgataaggat cgatccgggc
     6541 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     6601 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6661 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6721 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6781 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     6841 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6901 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     6961 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     7021 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     7081 tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg
     7141 cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag
     7201 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     7261 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     7321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     7381 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     7441 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     7501 tagtgaggag gcttttttgg aggatcaaac gacgccttgc aacaagctga gaaaaaacca
     7561 ggatcggcaa ggaaggattt tcaacggaaa cgttggcatc accgggtata aaaaggggcc
     7621 aatttttttg ttgatttgtt gcgcgaattc ttgctttcgg catcgaattc aagggagaag
     7681 gagtcgattc acacatacaa caagatccta ggcttttgca aaaagcttga ttcttctgac
     7741 acaacagtct cgaacttaag gctagagcca ccatgattga acaagatgga ttgcacgcag
     7801 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     7861 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     7921 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     7981 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     8041 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     8101 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     8161 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     8221 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     8281 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     8341 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     8401 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     8461 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     8521 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     8581 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat
     8641 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc
     8701 gcgtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     8761 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     8821 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     8881 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     8941 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     9001 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     9061 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     9121 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     9181 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     9241 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     9301 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     9361 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     9421 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     9481 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     9541 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     9601 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     9661 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     9721 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     9781 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     9841 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     9901 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     9961 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
    10021 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
    10081 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
    10141 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
    10201 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
    10261 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
    10321 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
    10381 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
    10441 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
    10501 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
    10561 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
    10621 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
    10681 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
    10741 tggccttttg ctggcctttt gctcacatgg ctcgac
//
