LOCUS       sequence-494371-        9753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For inverse prime editor using nCas9-D10A in HEK293T cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9753)
  AUTHORS   Liang R, Wang S, Cai Y, Li Z, Li KM, Wei J, Sun C, Zhu H, Chen K, 
            Gao C
  TITLE     Circular RNA-mediated inverse prime editing in human cells.
  JOURNAL   Nat Commun. 2025 May 31;16(1):5057. doi: 10.1038/s41467-025-59120-7.
  PUBMED    40447589
REFERENCE   2  (bases 1 to 9753)
  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: "2025-05-31"; volume: "16(1):5057. doi"; pages: " 
            10.1038/s41467-025-59120-7"
COMMENT     Sequence Label: iPE-D10A-Cas9
FEATURES             Location/Qualifiers
     source          1..9753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             694..714
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(715..738,739..741,742..4815)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             715..4815
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6988..7008
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7017..7034
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7057..7074)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7063..7287
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7358..7374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7358..7374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7371..7393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7382..7398
                     /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(7406..7412,7413..7430,7431..7436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7451..7472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7589..7606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7760..8348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7840..7859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8519..9310,9311..9379))
                     /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     9142..9161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9380..9484)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9563..9582)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcgacaag
      721 aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac
      781 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
      841 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
      901 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
      961 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     1021 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     1081 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     1141 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     1201 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     1261 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     1321 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     1381 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     1441 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     1501 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     1561 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     1621 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     1681 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     1741 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     1801 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     1861 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     1921 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     1981 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     2041 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     2101 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     2161 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     2221 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     2281 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     2341 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     2401 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     2461 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     2521 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     2581 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     2641 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     2701 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     2761 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     2821 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     2881 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     2941 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     3001 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     3061 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     3121 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     3181 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     3241 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     3301 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     3361 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     3421 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     3481 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     3541 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     3601 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     3661 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     3721 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     3781 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     3841 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     3901 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     3961 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     4021 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     4081 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     4141 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     4201 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     4261 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     4321 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     4381 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     4441 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     4501 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     4561 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     4621 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     4681 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     4741 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     4801 cagctgggag gtgactctgg aggatctagc ggaggatcct ctggcagcga gacaccagga
     4861 acaagcgagt cagcaacacc agagagcagt ggcggcagca gcggcggcag cagcacccta
     4921 aatatagaag atgagtatcg gctacatgag acctcaaaag agccagatgt ttctctaggg
     4981 tccacatggc tgtctgattt tcctcaggcc tgggcggaaa ccgggggcat gggactggca
     5041 gttcgccaag ctcctctgat catacctctg aaagcaacct ctacccccgt gtccataaaa
     5101 caatacccca tgtcacaaga agccagactg gggatcaagc cccacataca gagactgttg
     5161 gaccagggaa tactggtacc ctgccagtcc ccctggaaca cgcccctgct acccgttaag
     5221 aaaccaggga ctaatgatta taggcctgtc caggatctga gagaagtcaa caagcgggtg
     5281 gaagacatcc accccaccgt gcccaaccct tacaacctct tgagcgggct cccaccgtcc
     5341 caccagtggt acactgtgct tgatttaaag gatgcctttt tctgcctgag actccacccc
     5401 accagtcagc ctctcttcgc ctttgagtgg agagatccag agatgggaat ctcaggacaa
     5461 ttgacctgga ccagactccc acagggtttc aaaaacagtc ccaccctgtt taatgaggca
     5521 ctgcacagag acctagcaga cttccggatc cagcacccag acttgatcct gctacagtac
     5581 gtggatgact tactgctggc cgccacttct gagctagact gccaacaagg tactcgggcc
     5641 ctgttacaaa ccctagggaa cctcgggtat cgggcctcgg ccaagaaagc ccaaatttgc
     5701 cagaaacagg tcaagtatct ggggtatctt ctaaaagagg gtcagagatg gctgactgag
     5761 gccagaaaag agactgtgat ggggcagcct actccgaaga cccctcgaca actaagggag
     5821 ttcctaggga aggcaggctt ctgtcgcctc ttcatccctg ggtttgcaga aatggcagcc
     5881 cccctgtacc ctctcaccaa accggggact ctgtttaatt ggggcccaga ccaacaaaag
     5941 gcctatcaag aaatcaagca agctcttcta actgccccag ccctggggtt gccagatttg
     6001 actaagccct ttgaactctt tgtcgacgag aagcagggct acgccaaagg tgtcctaacg
     6061 caaaaactgg gaccttggcg tcggccggtg gcctacctgt ccaaaaagct agacccagta
     6121 gcagctgggt ggcccccttg cctacggatg gtagcagcca ttgccgtact gacaaaggat
     6181 gcaggcaagc taaccatggg acagccacta gtcattctgg ccccccatgc agtagaggca
     6241 ctagtcaaac aaccccccga ccgctggctt tccaacgccc ggatgactca ctatcaggcc
     6301 ttgcttttgg acacggaccg ggtccagttc ggaccggtgg tagccctgaa cccggctacg
     6361 ctgctcccac tgcctgagga agggctgcaa cacaactgcc ttgatatcct ggccgaagcc
     6421 cacggaaccc gacccgacct aacggaccag ccgctcccag acgccgacca cacctggtac
     6481 acggatggaa gcagtctctt acaagaggga cagcgtaagg cgggagctgc ggtgaccacc
     6541 gagaccgagg taatctgggc taaagccctg ccagccggga catccgctca gcgggctgaa
     6601 ctgatagcac tcacccaggc cctaaagatg gcagaaggta agaagctaaa tgtttatact
     6661 gatagccgtt atgcttttgc tactgcccat atccatggag aaatatacag aaggcgtggg
     6721 tggctcacat cagaaggcaa agagatcaaa aataaagacg agatcttggc cctactaaaa
     6781 gccctctttc tgcccaaaag acttagcata atccattgtc caggacatca aaagggacac
     6841 agcgccgagg ctagaggcaa ccggatggct gaccaagcgg cccgaaaggc agccatcaca
     6901 gagactccag acacctctac cctcctcata gaaaattcat caccctctgg cggctcaaaa
     6961 agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcta accggtcatc
     7021 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     7081 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     7141 ctgtcctttc ctaataaaat gagaaaattg catcgcattg tctgagtagg tgtcattcta
     7201 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     7261 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     7321 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     7381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7441 agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     8041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     8101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     8161 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     8221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     8281 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8341 gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg aacgaaaact
     8401 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8461 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8521 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8581 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8641 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8701 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8761 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8821 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8881 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8941 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     9001 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     9061 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     9121 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     9181 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     9241 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9301 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9361 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9481 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
     9541 atcccctagg gtctactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     9601 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     9661 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     9721 tgcttcgcga tgtacgggcc agatatacgc gtt
//
