LOCUS       sequence-415667-        9909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The CE1 construct, expressed from CMV or T7 promoters. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9909)
  AUTHORS   Ferreira da Silva J, Tou CJ, King EM, Eller ML, Rufino-Ramos D, Ma 
            L, Cromwell CR, Metovic J, Benning FMC, Chao LH, Eichler FS, 
            Kleinstiver BP
  TITLE     Click editing enables programmable genome writing using DNA 
            polymerases and HUH endonucleases.
  JOURNAL   Nat Biotechnol. 2024 Jul 22. doi: 10.1038/s41587-024-02324-x.
  PUBMED    39039307
REFERENCE   2  (bases 1 to 9909)
  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 
            Biotechnol. 2024 Jul 22. doi: 10.1038/s41587-024-02324-x."
COMMENT     Sequence Label: Click editor (CE1) - pCMV-T7-PCV2-nCas9-EcKlen...
FEATURES             Location/Qualifiers
     source          1..9909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(832..3345,3346..3348,3349..4932)
                     /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             832..4932
                     /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             4990..5010
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6895..6915
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6924..6941
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6964..6981)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6970..7194
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7265..7281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7265..7281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7278..7300)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7289..7305
                     /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(7313..7319,7320..7337,7338..7343))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7358..7379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7496..7513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7667..8255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7747..7766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8426..9217,9218..9286))
                     /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     9049..9068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9287..9391)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9470..9489)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(9661..9909,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gagcccatct
      421 aagaaaaacg gcagaagcgg acctcaacct cacaagcggt gggtctttac cctgaacaac
      481 cccagcgaag atgagagaaa gaagatccgg gacctgccta tcagcctgtt cgactacttc
      541 atcgtgggcg aggaaggaaa tgaggaaggc cggacccccc acctgcaggg cttcgccaac
      601 ttcgtgaaaa agcagacctt caacaaggtg aagtggtacc tgggcgctag atgccacatc
      661 gagaaggcca agggcaccga ccagcagaac aaagagtact gcagcaagga aggcaatctg
      721 ctgatggaat gtggcgcccc tagatcccag ggacagagaa gcggaggctc tagcggctcc
      781 gagacacccg gcacatctga gtccgccacc cctgagagca gcggcggcag cgacaagaag
      841 tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag
      901 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
      961 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg
     1021 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
     1081 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
     1141 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     1201 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     1261 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     1321 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     1381 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     1441 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     1501 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
     1561 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     1621 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     1681 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     1741 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     1801 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     1861 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
     1921 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     1981 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     2041 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     2101 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     2161 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     2221 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     2281 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     2341 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
     2401 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     2461 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     2521 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     2581 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     2641 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     2701 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     2761 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     2821 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     2881 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     2941 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     3001 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     3061 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     3121 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     3181 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     3241 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     3301 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggacgctat cgtgcctcag
     3361 agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg
     3421 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg
     3481 cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag
     3541 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     3601 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     3661 gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     3721 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     3781 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     3841 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     3901 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     3961 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     4021 acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg
     4081 aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     4141 ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag
     4201 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
     4261 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
     4321 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     4381 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     4441 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
     4501 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     4561 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac
     4621 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     4681 gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag
     4741 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     4801 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
     4861 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     4921 ctgggaggtg actccggcgg aagctctggt ggcagcaagc ggaccgccga cggctctgaa
     4981 ttcgagagcc ctaagaagaa aagaaaggtg agcggaggct ctagcggcgg aagcgttatc
     5041 tcctacgaca actacgtgac catcctggac gaagagacac tgaaggcttg gatcgctaaa
     5101 ctggaaaaag cccctgtgtt cgccttcgcc accgccaccg actccttaga taacatcagc
     5161 gccaacctgg tgggcctgtc tttcgccatc gagcctggcg tggccgccta catccccgtg
     5221 gcccacgatt acctggatgc cccagatcaa atcagcagag agagagcact ggagctcctg
     5281 aagcccctgc tggaggacga gaaggccctg aaggtgggac agaacctgaa atacgatagg
     5341 ggaatcctgg ctaattacgg catcgagctg agaggcatcg ccttcgatac catgctggaa
     5401 tcttacatcc tgaacagcgt ggccggccgc cacgacatgg acagcctggc tgaacggtgg
     5461 ctgaagcaca aaaccatcac cttcgaagag atcgccggaa agggcaagaa ccagctgaca
     5521 tttaatcaga tcgcactgga agaagccggc agatacgccg ctgaggacgc cgacgtgacc
     5581 cttcagctgc acctgaagat gtggcccgat ctgcagaagc acaagggccc actaaacgtg
     5641 ttcgagaaca tcgagatgcc tctggtgccc gtcctgagca gaatcgagag gaatggcgtg
     5701 aagatcgacc ccaaggtgct gcacaaccac agcgaggaac tgacactgcg gctggccgag
     5761 ctggaaaaga aggcccacga aatcgccggc gaggagttca acctgtctag caccaagcag
     5821 ctacaaacaa tcctgttcga gaagcagggc attaagcctc tgaaaaagac acctggcgga
     5881 gcccccagca ccagcgaaga agtgctggag gaactggcgc tggattaccc tctgcccaaa
     5941 gtgatcctgg aatatagagg actggccaag ctgaagtcta catacaccga caagctgcct
     6001 ctgatgatca accccaagac cggccgggtg catacatctt atcaccaggc cgtgacagcc
     6061 accggccggc tgagcagcac agatcctaac ctgcaaaaca tcccagtgcg gaacgaagag
     6121 ggcagaagaa tcagacaggc tttcattgcc cctgaggact acgtcatcgt gtctgctgac
     6181 tacagccaga tagaactgag aatcatggcc catctgtctc gggacaaggg cctgctgacc
     6241 gcttttgcag aaggcaagga catccacaga gccaccgctg ccgaagtgtt cggcctgcct
     6301 ctggaaacag tgaccagcga gcaaagaagg tccgccaagg ccattaactt cggcctgatc
     6361 tacggcatga gcgcctttgg cctggccaga cagctcaaca tccctagaaa ggaggcccag
     6421 aaatacatgg acctgtactt cgagcggtac cccggcgtgc tggagtacat ggaacggacc
     6481 agagcccagg ctaaagagca gggatacgtg gaaaccctgg atggccggag actgtatctg
     6541 cctgacatca agagcagcaa cggcgccaga agagccgccg ctgagagagc tgctatcaac
     6601 gcccctatgc agggcaccgc cgccgacatc atcaagcggg ccatgatcgc cgtggacgcc
     6661 tggctgcagg ccgagcagcc tagagtgcgg atgatcatgc aggtgcacga cgagctggtg
     6721 tttgaggtgc acaaggacga cgtggatgct gtggccaagc agattcacca gctgatggaa
     6781 aattgcacca gactggacgt gccactgctg gtcgaggttg gaagcggaga gaattgggac
     6841 caggcccact ctggcggctc aaaaagaacc gccgacggca gcgaattcga gtctcccaag
     6901 aagaagagga aagtctgacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     6961 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     7021 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     7081 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     7141 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     7201 aggcggaaag aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat
     7261 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7321 gagccggaag cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa
     7381 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7441 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     8041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     8101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     8161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     8221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8341 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8401 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8461 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8521 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgcgaa ccacgctcac
     8581 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8641 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8701 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8761 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8821 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8881 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8941 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     9001 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     9061 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     9121 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     9181 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     9241 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     9301 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9361 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     9421 tcgacggatc gggagatcga tctcccgatc ccctagggtc gactctcagt acaatctgct
     9481 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     9541 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     9601 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     9661 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     9721 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     9781 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     9841 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     9901 aagtgtatc
//
