LOCUS       sequence-350558-       11070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Delivers all prime editing nickase components in a single plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11070)
  AUTHORS   Adikusuma F, Lushington C, Arudkumar J, Godahewa GI, Chey YCJ, 
            Gierus L, Piltz S, Geiger A, Jain Y, Reti D, Wilson LOW, Bauer DC, 
            Thomas PQ
  TITLE     Optimized nickase- and nuclease-based prime editing in human and 
            mouse cells.
  JOURNAL   Nucleic Acids Res. 2021 Sep 17. pii: 6371974. doi: 
            10.1093/nar/gkab792.
  PUBMED    34534334
REFERENCE   2  (bases 1 to 11070)
  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: "Nucleic 
            Acids Res. 2021 Sep 17. pii: 6371974. doi: 10.1093/nar/gkab792."
COMMENT     Sequence Label: PEA1
FEATURES             Location/Qualifiers
     source          1..11070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          113..340
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      352..361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             361..426
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             433..453
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(478..2991,2992..2994,2995..4578)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             478..4578
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6751..6798
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(6826..6843)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6832..7039
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7048..7064
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7048..7064
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_RNA        complement(7107..7182)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(7200..7448)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     repeat_region   7457..7597
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7457..7586
                     /label=AAV2 ITR
     rep_origin      7672..8127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7759..7778)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7969..7990
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8144..8163)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8263..8285
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8323..8341)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8409..8513
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8514..8582,8583..9374)
                     /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(8732..8751)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9545..10133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10034..10053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        10195..10443
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        10462..10537
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        10622..10907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(10909..11070,1..112)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
       61 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      121 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
      181 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      241 taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt
      301 taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg
      361 gactataagg accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac
      421 gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac
      481 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc
      541 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
      601 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
      661 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
      721 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
      781 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
      841 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
      901 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
      961 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     1021 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     1081 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     1141 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     1201 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     1261 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     1321 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     1381 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     1441 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     1501 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     1561 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     1621 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     1681 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     1741 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     1801 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     1861 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     1921 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     1981 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     2041 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     2101 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     2161 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     2221 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     2281 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     2341 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     2401 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     2461 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     2521 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     2581 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     2641 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     2701 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     2761 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     2821 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     2881 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     2941 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga cgctatcgtg
     3001 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     3061 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     3121 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     3181 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     3241 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     3301 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     3361 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     3421 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     3481 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     3541 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     3601 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     3661 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     3721 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     3781 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     3841 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     3901 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     3961 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     4021 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     4081 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     4141 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     4201 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     4261 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     4321 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     4381 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     4441 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac gaaagaggtg
     4501 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     4561 tctcagctgg gaggtgactc tggaggatct agcggaggat cctctggcag cgagacacca
     4621 ggaacaagcg agtcagcaac accagagagc agtggcggca gcagcggcgg cagcagcacc
     4681 ctaaatatag aagatgagta tcggctacat gagacctcaa aagagccaga tgtttctcta
     4741 gggtccacat ggctgtctga ttttcctcag gcctgggcgg aaaccggggg catgggactg
     4801 gcagttcgcc aagctcctct gatcatacct ctgaaagcaa cctctacccc cgtgtccata
     4861 aaacaatacc ccatgtcaca agaagccaga ctggggatca agccccacat acagagactg
     4921 ttggaccagg gaatactggt accctgccag tccccctgga acacgcccct gctacccgtt
     4981 aagaaaccag ggactaatga ttataggcct gtccaggatc tgagagaagt caacaagcgg
     5041 gtggaggaca tccaccccac cgtgcccaac ccttacaacc tcttgagcgg gctcccaccg
     5101 tcccaccagt ggtacactgt gcttgattta aaggatgcct ttttctgcct gagactccac
     5161 cccaccagtc agcctctctt cgcctttgag tggagagatc cagagatggg aatctcagga
     5221 caattgacct ggaccagact cccacagggt ttcaaaaaca gtcccaccct gtttaatgag
     5281 gcactgcaca gagacctagc agacttccgg atccagcacc cagacttgat cctgctacag
     5341 tacgtggatg acttactgct ggccgccact tctgagctag actgccaaca aggtactcgg
     5401 gccctgttac aaaccctagg gaacctcggg tatcgggcct cggccaagaa agcccaaatt
     5461 tgccagaaac aggtcaagta tctggggtat cttctaaaag agggtcagag atggctgact
     5521 gaggccagaa aagagactgt gatggggcag cctactccaa agacccctcg acaactaagg
     5581 gagttcctag ggaaggcagg cttctgtcgc ctcttcatcc ctgggtttgc agaaatggca
     5641 gcccccctgt accctctcac caaaccgggg actctgttta attggggccc agaccaacaa
     5701 aaggcctatc aagaaatcaa gcaagccctt ctaactgccc cagccctggg gttgccagat
     5761 ttgactaagc cctttgaact ctttgtcgac gagaagcagg gctacgccaa aggtgtccta
     5821 acgcaaaaac tgggaccttg gcgtcggccg gtggcctacc tgtccaaaaa gctagaccca
     5881 gtagcagctg ggtggccccc ttgcctacgg atggtagcag ccattgccgt actgacaaag
     5941 gatgcaggca agctaaccat gggacagcca ctagtcattc tggcccccca tgcagtagag
     6001 gcactagtca aacaaccccc cgaccgctgg ctttccaacg cccggatgac tcactatcag
     6061 gccttgcttt tggacacgga ccgggtccag ttcggaccgg tggtagccct gaacccggct
     6121 acgctgctcc cactgcctga ggaagggctg caacacaact gccttgatat cctggccgaa
     6181 gcccacggaa cccgacccga cctaacggac cagccgctcc cagacgccga ccacacctgg
     6241 tacacggatg gaagcagtct cttacaagag ggacagcgta aggcgggagc tgcggtgacc
     6301 accgagaccg aggtaatctg ggctaaagcc ctgccagccg ggacatccgc tcagcgggct
     6361 gaactgatag cactcaccca ggccctaaag atggcagaag gtaagaagct aaatgtttat
     6421 actgatagcc gttatgcttt tgctactgcc catatccatg gagaaatata cagaaggcgt
     6481 gggtggctca catcagaagg caaagagatc aaaaataaag acgagatctt ggccctacta
     6541 aaagccctct ttctgcccaa aagacttagc ataatccatt gtccaggaca tcaaaaggga
     6601 cacagcgccg aggctagagg caaccggatg gctgaccaag cggcccgaaa ggcagccatc
     6661 acagagactc cagacacctc taccctcctc atagaaaatt catcaccctc tggcggctca
     6721 aaaagaaccg ccgacggcag cgaattcgag aaaaggccgg cggccacgaa aaaggccggc
     6781 caggcaaaaa agaaaaagta agaattccta gagctcgctg atcagcctcg actgtgcctt
     6841 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     6901 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     6961 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga
     7021 atagcaggca tgctggggag cggccgctct agaactagtg gatcccccgg gctgcaggaa
     7081 ttcgatacga atcctgcagg aaaaaagcac cgactcggtg ccactttttc aagttgataa
     7141 cggactagcc ttattttaac ttgctatttc tagctctaaa acgtcttctc gaagaccccg
     7201 gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt tcaagttacg
     7261 gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta cccaagaaat
     7321 tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt caaattaatt
     7381 ccaattatct ctctaacagc cttgtatcgt atatgcaaat atgaaggaat catgggaaat
     7441 aggccctcgc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     7501 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     7561 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     7621 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     7681 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     7741 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     7801 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     7861 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     7921 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     7981 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     8041 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     8101 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     8161 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     8221 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     8281 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     8341 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     8401 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     8461 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     8521 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     8581 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     8641 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     8701 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     8761 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     8821 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     8881 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     8941 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     9001 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     9061 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     9121 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     9181 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta
     9241 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     9301 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     9361 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     9421 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     9481 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     9541 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     9601 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     9661 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     9721 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     9781 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     9841 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     9901 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     9961 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
    10021 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
    10081 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
    10141 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtgagggc
    10201 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
    10261 ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
    10321 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
    10381 accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac
    10441 accgggtctt cgagaagacc tgttttagag ctagaaatag caagttaaaa taaggctagt
    10501 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcggg tcttcgagaa gacctttttt
    10561 ttgttctagt ccgtttttag cgcgtgcgcc aattctgcag acaaatggct ctagaggtac
    10621 ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
    10681 tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
    10741 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
    10801 acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc ttatgggact
    10861 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
    10921 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
    10981 ttatttttta attattttgt gcagcgatgg gggcgggggg ggggggcgcg cgccaggcgg
    11041 ggcggggcgg ggcgaggggc ggggcggggc
//
