LOCUS       sequence-430349-        9784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of nuclear nCas9n.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9784)
  AUTHORS   Chaumont L, Jouneau L, Huetz F, van Muilekom DR, Peruzzi M, Raffy C,
            Le Hir J, Minke J, Boudinot P, Collet B
  TITLE     Unexpected regulatory functions of cyprinid Viperin on inflammation 
            and metabolism.
  JOURNAL   BMC Genomics. 2024 Jun 29;25(1):650. doi: 
            10.1186/s12864-024-10566-x.
  PUBMED    38951796
REFERENCE   2  (bases 1 to 9784)
  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: "BMC 
            Genomics."; date: "2024-06-29"; volume: "25(1):650. doi"; pages: " 
            10.1186/s12864-024-10566-x"
COMMENT     Sequence Label: pcDNA3.1-Hyg-nCas9n
FEATURES             Location/Qualifiers
     source          1..9784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        918..936
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     918..935
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      982..991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1000..1020
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1027..5127
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes MGAS15252 Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRL
                     ENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             5040..5048
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             5143..5163
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5206..5223)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5212..5436
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5482..5910
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5569..5588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5779..5800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5919..5939)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5924..6253
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6104..6239
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6166..6185
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6302..7327
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6302..7327
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    7457..7578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7494..7513)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7548..7567
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7627..7643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7627..7643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7640..7662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7651..7667
                     /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(7675..7681,7682..7699,7700..7705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7720..7741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7858..7875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8029..8617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8109..8128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8788..9579,9580..9648))
                     /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     9411..9430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9649..9753)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgatt taggtgacac tatagaatac aagctacttg ttctttttgc
      961 aggatccgat ccaatgtcga cgccaccatg gcttctccac ctaagaagaa gagaaaggtg
     1021 ggaagcatgg ataagaagta tagcatcggc ctggatattg gaactaactc cgtgggttgg
     1081 gcagtgatta cagacgacta caaggtccct agcaagaaat ttaaggtgct gggtaacacc
     1141 gacaggcaca gcatcaagaa aaatctgatt ggagccctgc tgttcggttc tggagagact
     1201 gccgaagcaa cacgcctgaa aagaacagca agaaggcgct ataccagaag gaagaataga
     1261 atctgttacc tgcaggagat tttctctaac gaaatggcta aggtggacga ttcattcttt
     1321 cataggctgg aggaaagttt cctggtcgag gaagataaga aacacgagcg ccatcctatc
     1381 tttggaaaca ttgtggacga ggtcgcctat cacgaaaaat acccaaccat ctatcatctg
     1441 cgcaagaaac tggctgactc tactgataaa gccgacctga gactgatcta tctggctctg
     1501 gcccacatga ttaagttcag gggtcatttt ctgatcgagg gcgatctgaa ccccgacaat
     1561 tccgatgtgg acaagctgtt catccagctg gtccagattt acaatcagct gtttgaggaa
     1621 aaccctatta atgcttccag agtggacgca aaagctatcc tgtcagccag gctgtccaag
     1681 tcacgcagac tggagaacct gattgcacag ctgcccggag aaaagaggaa cggtctgttt
     1741 ggaaatctga tcgctctgag tctgggcctg actcctaact tcaaaagcaa ttttgatctg
     1801 gctgaggacg ccaaactgca gctgtcaaag gacacatatg acgatgacct ggataacctg
     1861 ctggcacaga tcggagatca gtacgctgac ctgttcctgg ctgccaaaaa tctgtccgac
     1921 gcaatcctgc tgtcagatat tctgagagtg aacagcgaga ttacaaaagc tcctctgagt
     1981 gccagcatga tcaagagata tgacgagcac catcaggatc tgaccctgct gaaggctctg
     2041 gtcaggcagc agctgccaga gaagtacaag gaaattttct ttgatcagtc caagaacggc
     2101 tacgccggtt atatcgacgg aggcgcatca caggaggaat tctacaagtt tatcaaacct
     2161 attctggaga agatggacgg aactgaggaa ctgctggtga aactgaatag agaggacctg
     2221 ctgaggaagc agcgcacatt tgataacggt tccatcccac accagattca tctgggagag
     2281 ctgcacgcta tcctgaggcg ccaggaagac ttctacccct ttctgaaaga taaccgcgag
     2341 aagatcgaaa aaattctgac cttcagaatc ccttactatg tgggtccact ggctcgcgga
     2401 aacagcagat ttgcctggat gactcgcaaa tccgaggaaa ccattactcc ttggaacttc
     2461 gaggaagtgg tcgataaggg cgcctctgca cagtccttca tcgagagaat gactaatttt
     2521 gacaaaaacc tgcccaatga gaaagtgctg cctaagcact ccctgctgta cgagtatttc
     2581 actgtctata acgaactgac aaaggtgaaa tacgtcaccg agggcatgag aaagccagcc
     2641 ttcctgtcag gagagcagaa gaaagcaatc gtggatctgc tgtttaaaac caacaggaaa
     2701 gtgactgtca agcagctgaa ggaggactac ttcaagaaaa ttgaatgctt cgattccgtg
     2761 gagatcagcg gagtcgaaga cagatttaac gcaagcctgg gcgcttacca cgatctgctg
     2821 aagatcatta aggataaaga cttcctggac aacgaggaaa atgaggatat cctggaagac
     2881 attgtgctga cactgaccct gtttgaggac agaggaatga tcgaggaaag actgaaaacc
     2941 tatgctcatc tgttcgatga caaggtgatg aaacagctga agagaaggcg ctacactggc
     3001 tggggtagac tgagcaggaa gctgatcaac ggcattaggg ataaacagtc aggaaagaca
     3061 atcctggact ttctgaaaag tgatggcttc gccaaccgca attttatgca gctgattcac
     3121 gatgacagtc tgaccttcaa agaggacatc cagaaggctc aggtgtctgg acagggccac
     3181 tccctgcatg agcagattgc aaacctggct ggaagcccag ccatcaagaa aggcattctg
     3241 cagacagtga aaatcgtcga tgagctggtg aaagtcatgg gccataagcc cgaaaacatc
     3301 gtgattgaga tggctcgcga aaatcagaca acccagaagg gtcagaagaa cagtagagag
     3361 aggatgaaaa gaatcgagga aggcattaag gagctgggta gccagatcct gaaagagcac
     3421 ccagtggaaa acacacagct gcagaatgag aagctgtatc tgtactatct gcagaatgga
     3481 agagatatgt acgtggacca ggagctggat attaacaggc tgtctgatta cgacgtggat
     3541 catatcgtcc cccagagttt catcaaagat gacagcattg acaacaaggt gctgaccagg
     3601 tccgacaaaa acagaggaaa atcagataat gtccctagtg aggaagtggt caagaaaatg
     3661 aagaactact ggagacagct gctgaatgcc aaactgatca ctcagaggaa gtttgataac
     3721 ctgacaaaag cagagcgcgg tggactgtca gaactggaca aagctggatt catcaagagg
     3781 cagctggtgg aaacacgcca gatcactaaa cacgtcgcac agattctgga tagtcgcatg
     3841 aacacaaagt acgatgagaa tgacaaactg atcagagaag tgaaggtcat taccctgaag
     3901 agtaaactgg tcagcgactt taggaaagat ttccagtttt ataaggtccg cgagattaac
     3961 aattatcacc atgcccatga cgcatacctg aacgccgtgg tcggtaccgc actgatcaag
     4021 aaatacccaa aactggagag cgaattcgtg tacggagact ataaggtgta cgatgtcaga
     4081 aaaatgatcg ccaagtccga gcaggaaatt ggaaaagcta ctgccaagta tttcttttac
     4141 tcaaacatca tgaatttctt taagacagag atcaccctgg ccaatggaga aatccgcaaa
     4201 aggcccctga ttgagacaaa cggagagaca ggcgaaatcg tgtgggacaa aggcagagat
     4261 tttgcaaccg tgaggaaggt cctgagcatg cctcaagtga atatcgtcaa gaaaactgag
     4321 gtgcagacag gcggtttctc aaaagaaagt attctgccaa aacgcaactc tgataagctg
     4381 atcgctagaa agaaagactg ggaccctaag aagtatggag gctttgactc tcccactgtg
     4441 gcatactccg tcctggtggt cgctaaggtg gagaagggca aaagcaagaa actgaaatct
     4501 gtcaaggagc tgctgggtat cacaattatg gagagaagct ctttcgagaa gaacccaatc
     4561 gattttctgg aggccaaagg ttataaggaa gtgaagaaag acctgatcat taaactgccc
     4621 aagtacagtc tgtttgagct ggaaaacggc aggaaacgca tgctggcaag cgctggagag
     4681 ctgcagaaag gcaatgaact ggccctgcct tctaagtacg tgaacttcct gtatctggca
     4741 agccactacg agaagctgaa aggatctcca gaggataacg aacagaaaca gctgtttgtg
     4801 gagcagcaca agcattatct ggacgagatc attgaacaga ttagcgagtt ctctaaaaga
     4861 gtgatcctgg ccgacgcaaa tctggataag gtcctgtctg cttacaacaa acacagagat
     4921 aagcccatca gggagcaggc cgaaaatatc attcatctgt tcactctgac aaacctgggc
     4981 gcacctgcag ctttcaagta cttcgacact acaatcgata gaaagaggta cacctccact
     5041 aaggaggtgc tggacgctac actgatccat cagagtatta ccggcctgta cgaaacaagg
     5101 attgacctgt ctcagctggg tggcgactca ccggtgagat ctcctaagaa gaagagaaag
     5161 gtgtgaccgc ggaatctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     5221 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     5281 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     5341 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     5401 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     5461 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5521 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5581 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5641 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5701 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5761 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5821 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5881 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     5941 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     6001 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6061 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     6121 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6181 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6241 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     6301 gatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     6361 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     6421 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     6481 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     6541 tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     6601 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga
     6661 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     6721 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     6781 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     6841 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     6901 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     6961 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     7021 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     7081 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     7141 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     7201 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
     7261 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     7321 ggaatagcac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     7381 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     7441 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7501 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7561 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     7621 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7681 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7741 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7801 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7861 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7921 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7981 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     8041 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     8101 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     8161 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     8221 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     8281 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8341 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8401 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8461 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8521 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8581 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8641 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8701 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8761 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8821 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8881 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8941 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     9001 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     9061 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     9121 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     9181 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     9241 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     9301 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9361 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9421 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9481 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9541 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9601 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9661 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9721 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9781 cgtc
//
