LOCUS       sequence-391374-        9443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct encoding SpCas9 nuclease and a control 
            non-targeting guide RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9443)
  AUTHORS   Wang Q, Liu J, Janssen JM, Le Bouteiller M, Frock RL, Goncalves MAFV
  TITLE     Precise and broad scope genome editing based on high-specificity 
            Cas9 nickases.
  JOURNAL   Nucleic Acids Res. 2021 Jan 25;49(2):1173-1198. doi: 
            10.1093/nar/gkaa1236.
  PUBMED    33398349
REFERENCE   2  (bases 1 to 9443)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2021-01-25- 25"; volume: "49"; issue: 
            "2"; pages: "1173-1198"
COMMENT     Sequence Label: AV13_pCAG.Cas9.gRNA.NT
FEATURES             Location/Qualifiers
     source          1..9443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        225..473
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     225..245
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     396..415
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        494..569
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        755..1134
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1136..1411
                     /label=chicken beta-actin promoter
     primer_bind     2341..2363
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2425..2444
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2515..2524
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2521..6624
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2521..6624
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2521..6624
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6637..6657
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6712..6731)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6756..6804
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(7250..7268)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7323..7341)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7409..7513
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7514..7582,7583..8374)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7732..7751)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8545..9133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9034..9053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9287..9304
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     9371..9387
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9394..9413
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9394..9412
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atgttattcc tccttattta aaagcttaag gtgcacggcc cacgtggcca ctagtacttc
       61 tcgagctctg tacatgtccg cggtcgcgac gtacgcgtat cgatggcgcc agctgcaggc
      121 ggccgccata tgcatcctag gcctattaat attccggagt atacgtagcc ggctaacgtt
      181 atcgcatgcc tgcagagctc tagagtccgg ccgccccctt caccgagggc ctatttccca
      241 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
      301 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
      361 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
      421 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accggtgagc
      481 tcttatttgc gtagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
      541 aacttgaaaa agtggcaccg agtcggtgct ttttttgaat taacaaccgg tacctctaga
      601 actatagcta gcatgcgcaa atttaaagcg ctgatatcga tcgcgcgcag atctgtcatg
      661 atgatcattg caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
      721 atgtacgggc cagatatact cgagccatgg attcgacatt gattattgac tagttattaa
      781 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      841 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      901 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      961 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     1021 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1081 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
     1141 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     1201 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggsgcgc
     1261 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
     1321 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     1381 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
     1441 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
     1501 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
     1561 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt
     1621 gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     1681 cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct
     1741 ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg
     1801 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
     1861 cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg agcacggccc
     1921 ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgg tggcggcagg
     1981 tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg
     2041 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
     2101 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
     2161 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
     2221 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
     2281 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     2341 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     2401 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     2461 gcaaagaatt atcgcatgcc tgcagagctc tagagtctag aggatcgaac ccttgccacc
     2521 atggacaaga agtactccat tgggctcgat atcggcacaa acagcgtcgg ctgggccgtc
     2581 attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc
     2641 cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga gacggccgaa
     2701 gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa tcggatctgc
     2761 tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt cttccatagg
     2821 ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc aatctttggc
     2881 aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca tctgaggaag
     2941 aagcttgtag acagtactga taaggctgac ttgcggttga tctatctcgc gctggcgcat
     3001 atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga caacagcgat
     3061 gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga agagaacccg
     3121 atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc caaatcccgg
     3181 cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct gtttggtaat
     3241 cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga cctggccgaa
     3301 gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa tctgctggcc
     3361 cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc agacgccatt
     3421 ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct gagcgctagt
     3481 atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga
     3541 cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc
     3601 ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa gcccatcttg
     3661 gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga tctgttgcgc
     3721 aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg cgaactgcac
     3781 gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag ggaaaagatt
     3841 gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg gggaaattcc
     3901 agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa cttcgaggaa
     3961 gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa
     4021 aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta cttcacagtt
     4081 tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc agcattcctg
     4141 tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg gaaagttacc
     4201 gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc tgttgaaatc
     4261 agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct cctgaaaatc
     4321 attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga ggacattgtc
     4381 ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa aacttacgct
     4441 catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac aggatggggg
     4501 cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa gacaatcctg
     4561 gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat ccatgatgac
     4621 tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg ggacagtctt
     4681 cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat actgcagacc
     4741 gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga gaatatcgtt
     4801 atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag tagggaaagg
     4861 atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa ggaacaccca
     4921 gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca gaacggcagg
     4981 gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga cgtggatcat
     5041 atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt gacaagatcc
     5101 gataaaaata gagggaagag tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa
     5161 aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt cgataatctg
     5221 actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat caaaaggcag
     5281 cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc acgcatgaac
     5341 accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac tctgaagtct
     5401 aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga gatcaacaat
     5461 taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact tatcaaaaaa
     5521 tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga tgttaggaaa
     5581 atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt cttttacagc
     5641 aatattatga attttttcaa gaccgagatt acactggcca atggagagat tcggaagcga
     5701 ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg tagggatttc
     5761 gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta
     5821 cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc
     5881 gcacgcaaaa aagattggga ccccaagaaa tacggcggat tcgattctcc tacagtcgct
     5941 tacagtgtac tggttgtggc caaagtggag aaagggaagt ctaaaaaact caaaagcgtc
     6001 aaggaactgc tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa ccccatcgac
     6061 tttctcgagg cgaaaggata taaagaggtc aaaaaagacc tcatcattaa gcttcccaag
     6121 tactctctct ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg
     6181 cagaaaggta acgagctggc actgccctct aaatacgtta atttcttgta tctggccagc
     6241 cactatgaaa agctcaaagg gtctcccgaa gataatgagc agaagcagct gttcgtggaa
     6301 caacacaaac actaccttga tgagatcatc gagcaaataa gcgaattctc caaaagagtg
     6361 atcctcgccg acgctaacct cgataaggtg ctttctgctt acaataagca cagggataag
     6421 cccatcaggg agcaggcaga aaacattatc cacttgttta ctctgaccaa cttgggcgcg
     6481 cctgcagcct tcaagtactt cgacaccacc atagacagaa agcggtacac ctctacaaag
     6541 gaggtcctgg acgccacact gattcatcag tcaattacgg ggctctatga aacaagaatc
     6601 gacctctctc agctcggtgg agacagcagg gctgacccca agaagaagag gaaggtgtga
     6661 aagggttcga tccctaccgg ttagtaatga gtttaaacgg gggaggctaa ctgaaacacg
     6721 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     6781 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     6841 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     6901 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     6961 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     7021 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     7081 ccctagcgcc cggggttata attacctcag gtcgacgtcc catgtgcagg tgctgaattc
     7141 gaaggcattc cacgacgtgg tcaccagcca tggccatgtc caactccatc aaatcagctt
     7201 gagtagccat gcccatggct ccagcctgtc ctcgtaccag cttttgttcc ctttagtgag
     7261 ggttaatttc gagcttggcg taatcatggt catagctgtt tccaattctt gaagacgaaa
     7321 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     7381 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     7441 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     7501 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7561 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7621 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7681 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7741 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     7801 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7861 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7921 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7981 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     8041 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     8101 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8161 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8221 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8281 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     8341 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     8401 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     8461 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8521 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     8581 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     8641 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     8701 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8761 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8821 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8881 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8941 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     9001 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     9061 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     9121 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     9181 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     9241 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     9301 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     9361 acaccgcata gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg
     9421 gagcttggcg taatcatggt cat
//
