LOCUS       sequence-432858-        9891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Variant CE1 construct with GoCV HUH endonuclease, expressed from CMV
            or T7 promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9891)
  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 9891)
  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: HUHe variant click editor (CE1) -
            pCMV-T7-GoCV...
FEATURES             Location/Qualifiers
     source          1..9891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(113..217)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(296..315)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        487..866
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        867..1070
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1021..1041
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1112..1131
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1112..1130
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1138..1147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1549..4062,4063..4065,4066..5649)
                     /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             1549..5649
                     /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             5707..5727
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7612..7632
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7641..7658
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7681..7698)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7687..7911
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7982..7998)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7982..7998)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7995..8017)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8006..8022
                     /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(8030..8036,8037..8054,8055..8060))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8075..8096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8213..8230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8384..8972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8464..8483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9143..9891,1..43,44..112))
                     /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     9766..9785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
       61 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      181 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      241 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
      301 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      361 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      421 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      481 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      541 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      601 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      661 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      721 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      781 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      841 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      901 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      961 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     1021 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     1081 accgtcagat ccgctagaga tccgcggccg ctaatacgac tcactatagg gagagccgcc
     1141 accatggcca aaaacggcaa ctacagctac aagcggtggg tgttcaccat caacaacccc
     1201 acattcgagg actactgcgc cgtggtggaa ttctgcaacc tcgacaattg caagttcgcc
     1261 atcgtgggag aggaaaaggg cgagaaagag ggcacccctc acctgcaggg ctttctgagc
     1321 ctgagaaaga acgccaaggc cgctgctctg gaagagaacc tgggcggaag agcctggctg
     1381 tctagagcca gaggcagcga cgaggataat gaggaatatt gtagcaagga atccacctac
     1441 ctgcgggtcg gcgagcctaa ccggaagggc agaagcagcg gaggctctag cggctccgag
     1501 acacccggca catctgagtc cgccacccct gagagcagcg gcggcagcga caagaagtac
     1561 agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
     1621 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
     1681 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
     1741 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
     1801 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
     1861 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
     1921 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
     1981 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
     2041 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     2101 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     2161 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     2221 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     2281 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     2341 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     2401 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     2461 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     2521 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     2581 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     2641 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     2701 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     2761 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     2821 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     2881 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     2941 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     3001 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     3061 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     3121 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     3181 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     3241 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     3301 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     3361 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     3421 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     3481 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     3541 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     3601 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     3661 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     3721 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     3781 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     3841 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     3901 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     3961 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     4021 caggaactgg acatcaaccg gctgtccgac tacgatgtgg acgctatcgt gcctcagagc
     4081 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     4141 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     4201 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     4261 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     4321 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     4381 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     4441 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     4501 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     4561 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     4621 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     4681 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     4741 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     4801 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     4861 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     4921 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     4981 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     5041 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     5101 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     5161 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     5221 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     5281 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     5341 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     5401 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     5461 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     5521 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     5581 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     5641 ggaggtgact ccggcggaag ctctggtggc agcaagcgga ccgccgacgg ctctgaattc
     5701 gagagcccta agaagaaaag aaaggtgagc ggaggctcta gcggcggaag cgttatctcc
     5761 tacgacaact acgtgaccat cctggacgaa gagacactga aggcttggat cgctaaactg
     5821 gaaaaagccc ctgtgttcgc cttcgccacc gccaccgact ccttagataa catcagcgcc
     5881 aacctggtgg gcctgtcttt cgccatcgag cctggcgtgg ccgcctacat ccccgtggcc
     5941 cacgattacc tggatgcccc agatcaaatc agcagagaga gagcactgga gctcctgaag
     6001 cccctgctgg aggacgagaa ggccctgaag gtgggacaga acctgaaata cgatagggga
     6061 atcctggcta attacggcat cgagctgaga ggcatcgcct tcgataccat gctggaatct
     6121 tacatcctga acagcgtggc cggccgccac gacatggaca gcctggctga acggtggctg
     6181 aagcacaaaa ccatcacctt cgaagagatc gccggaaagg gcaagaacca gctgacattt
     6241 aatcagatcg cactggaaga agccggcaga tacgccgctg aggacgccga cgtgaccctt
     6301 cagctgcacc tgaagatgtg gcccgatctg cagaagcaca agggcccact aaacgtgttc
     6361 gagaacatcg agatgcctct ggtgcccgtc ctgagcagaa tcgagaggaa tggcgtgaag
     6421 atcgacccca aggtgctgca caaccacagc gaggaactga cactgcggct ggccgagctg
     6481 gaaaagaagg cccacgaaat cgccggcgag gagttcaacc tgtctagcac caagcagcta
     6541 caaacaatcc tgttcgagaa gcagggcatt aagcctctga aaaagacacc tggcggagcc
     6601 cccagcacca gcgaagaagt gctggaggaa ctggcgctgg attaccctct gcccaaagtg
     6661 atcctggaat atagaggact ggccaagctg aagtctacat acaccgacaa gctgcctctg
     6721 atgatcaacc ccaagaccgg ccgggtgcat acatcttatc accaggccgt gacagccacc
     6781 ggccggctga gcagcacaga tcctaacctg caaaacatcc cagtgcggaa cgaagagggc
     6841 agaagaatca gacaggcttt cattgcccct gaggactacg tcatcgtgtc tgctgactac
     6901 agccagatag aactgagaat catggcccat ctgtctcggg acaagggcct gctgaccgct
     6961 tttgcagaag gcaaggacat ccacagagcc accgctgccg aagtgttcgg cctgcctctg
     7021 gaaacagtga ccagcgagca aagaaggtcc gccaaggcca ttaacttcgg cctgatctac
     7081 ggcatgagcg cctttggcct ggccagacag ctcaacatcc ctagaaagga ggcccagaaa
     7141 tacatggacc tgtacttcga gcggtacccc ggcgtgctgg agtacatgga acggaccaga
     7201 gcccaggcta aagagcaggg atacgtggaa accctggatg gccggagact gtatctgcct
     7261 gacatcaaga gcagcaacgg cgccagaaga gccgccgctg agagagctgc tatcaacgcc
     7321 cctatgcagg gcaccgccgc cgacatcatc aagcgggcca tgatcgccgt ggacgcctgg
     7381 ctgcaggccg agcagcctag agtgcggatg atcatgcagg tgcacgacga gctggtgttt
     7441 gaggtgcaca aggacgacgt ggatgctgtg gccaagcaga ttcaccagct gatggaaaat
     7501 tgcaccagac tggacgtgcc actgctggtc gaggttggaa gcggagagaa ttgggaccag
     7561 gcccactctg gcggctcaaa aagaaccgcc gacggcagcg aattcgagtc tcccaagaag
     7621 aagaggaaag tctgaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     7681 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     7741 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     7801 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     7861 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     7921 cggaaagaac cagctggggc tcgataccgt cgacctctag ctagagcttg gcgtaatcat
     7981 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     8041 ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt gagctaactc acattaattg
     8101 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     8161 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     8221 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     8281 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     8341 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     8401 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8461 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     8521 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     8581 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8641 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8701 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8761 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8821 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8881 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     8941 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     9001 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     9061 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     9121 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     9181 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     9241 gggagggctt accatctggc cccagtgctg caatgatacc gcgcgaacca cgctcaccgg
     9301 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     9361 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     9421 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     9481 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     9541 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     9601 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     9661 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     9721 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     9781 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     9841 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc a
//
