LOCUS       sequence-415740-        9930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A variant CE1 construct with TGMV HUH enconuclease, expressed from 
            CMV or T7 promoters. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9930)
  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 9930)
  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-TGMV...
FEATURES             Location/Qualifiers
     source          1..9930
                     /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"
     CDS             join(1588..4101,4102..4104,4105..5688)
                     /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             1588..5688
                     /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             5746..5766
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7651..7671
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7680..7697
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7720..7737)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7726..7950
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8021..8037)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8021..8037)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8034..8056)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8045..8061
                     /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(8069..8075,8076..8093,8094..8099))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8114..8135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8252..8269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8423..9011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8503..8522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9182..9930,1..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     9805..9824
                     /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 accatgccta gccaccctaa gcggtttcag atcaatgcca agaactactt cctgacctac
     1201 ccccagtgca gcctcagcaa ggaagagagc ctgtctcagc tgcaggccct gaacacccct
     1261 attaacaaga aattcatcaa gatctgcaga gagctgcatg aagatggcca gcctcacctg
     1321 cacgtgctga tccaatttga gggcaagtac tgctgtcaga accagagatt cttcgacctg
     1381 gtgtccccaa caagatctgc tcacttccac cccaacatcc agcgggccaa aagctccagc
     1441 gacgtgaaga cctatatcga caaggacgga gatacactgg tctggggcga gttccaagtg
     1501 gacggcagaa gcgccagcgg aggctctagc ggctccgaga cacccggcac atctgagtcc
     1561 gccacccctg agagcagcgg cggcagcgac aagaagtaca gcatcggcct ggacatcggc
     1621 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     1681 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     1741 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     1801 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     1861 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     1921 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     1981 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     2041 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     2101 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     2161 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     2221 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     2281 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     2341 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     2401 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     2461 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     2521 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     2581 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     2641 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     2701 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     2761 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     2821 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     2881 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     2941 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     3001 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     3061 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     3121 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     3181 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     3241 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     3301 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     3361 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     3421 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     3481 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     3541 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     3601 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     3661 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     3721 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     3781 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     3841 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     3901 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     3961 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     4021 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     4081 ctgtccgact acgatgtgga cgctatcgtg cctcagagct ttctgaagga cgactccatc
     4141 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     4201 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     4261 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     4321 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     4381 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     4441 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     4501 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     4561 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     4621 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     4681 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     4741 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     4801 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     4861 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     4921 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     4981 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     5041 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     5101 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     5161 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     5221 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     5281 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     5341 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     5401 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     5461 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     5521 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     5581 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     5641 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggtgactc cggcggaagc
     5701 tctggtggca gcaagcggac cgccgacggc tctgaattcg agagccctaa gaagaaaaga
     5761 aaggtgagcg gaggctctag cggcggaagc gttatctcct acgacaacta cgtgaccatc
     5821 ctggacgaag agacactgaa ggcttggatc gctaaactgg aaaaagcccc tgtgttcgcc
     5881 ttcgccaccg ccaccgactc cttagataac atcagcgcca acctggtggg cctgtctttc
     5941 gccatcgagc ctggcgtggc cgcctacatc cccgtggccc acgattacct ggatgcccca
     6001 gatcaaatca gcagagagag agcactggag ctcctgaagc ccctgctgga ggacgagaag
     6061 gccctgaagg tgggacagaa cctgaaatac gataggggaa tcctggctaa ttacggcatc
     6121 gagctgagag gcatcgcctt cgataccatg ctggaatctt acatcctgaa cagcgtggcc
     6181 ggccgccacg acatggacag cctggctgaa cggtggctga agcacaaaac catcaccttc
     6241 gaagagatcg ccggaaaggg caagaaccag ctgacattta atcagatcgc actggaagaa
     6301 gccggcagat acgccgctga ggacgccgac gtgacccttc agctgcacct gaagatgtgg
     6361 cccgatctgc agaagcacaa gggcccacta aacgtgttcg agaacatcga gatgcctctg
     6421 gtgcccgtcc tgagcagaat cgagaggaat ggcgtgaaga tcgaccccaa ggtgctgcac
     6481 aaccacagcg aggaactgac actgcggctg gccgagctgg aaaagaaggc ccacgaaatc
     6541 gccggcgagg agttcaacct gtctagcacc aagcagctac aaacaatcct gttcgagaag
     6601 cagggcatta agcctctgaa aaagacacct ggcggagccc ccagcaccag cgaagaagtg
     6661 ctggaggaac tggcgctgga ttaccctctg cccaaagtga tcctggaata tagaggactg
     6721 gccaagctga agtctacata caccgacaag ctgcctctga tgatcaaccc caagaccggc
     6781 cgggtgcata catcttatca ccaggccgtg acagccaccg gccggctgag cagcacagat
     6841 cctaacctgc aaaacatccc agtgcggaac gaagagggca gaagaatcag acaggctttc
     6901 attgcccctg aggactacgt catcgtgtct gctgactaca gccagataga actgagaatc
     6961 atggcccatc tgtctcggga caagggcctg ctgaccgctt ttgcagaagg caaggacatc
     7021 cacagagcca ccgctgccga agtgttcggc ctgcctctgg aaacagtgac cagcgagcaa
     7081 agaaggtccg ccaaggccat taacttcggc ctgatctacg gcatgagcgc ctttggcctg
     7141 gccagacagc tcaacatccc tagaaaggag gcccagaaat acatggacct gtacttcgag
     7201 cggtaccccg gcgtgctgga gtacatggaa cggaccagag cccaggctaa agagcaggga
     7261 tacgtggaaa ccctggatgg ccggagactg tatctgcctg acatcaagag cagcaacggc
     7321 gccagaagag ccgccgctga gagagctgct atcaacgccc ctatgcaggg caccgccgcc
     7381 gacatcatca agcgggccat gatcgccgtg gacgcctggc tgcaggccga gcagcctaga
     7441 gtgcggatga tcatgcaggt gcacgacgag ctggtgtttg aggtgcacaa ggacgacgtg
     7501 gatgctgtgg ccaagcagat tcaccagctg atggaaaatt gcaccagact ggacgtgcca
     7561 ctgctggtcg aggttggaag cggagagaat tgggaccagg cccactctgg cggctcaaaa
     7621 agaaccgccg acggcagcga attcgagtct cccaagaaga agaggaaagt ctgaccggtc
     7681 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     7741 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     7801 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     7861 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     7921 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     7981 cgataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     8041 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     8101 cctagggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     8161 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     8221 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     8281 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     8341 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     8401 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     8461 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8521 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8581 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8641 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8701 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8761 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8821 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     8881 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8941 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     9001 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     9061 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     9121 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     9181 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     9241 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     9301 ccagtgctgc aatgataccg cgcgaaccac gctcaccggc tccagattta tcagcaataa
     9361 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     9421 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9481 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9541 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9601 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9661 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9721 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9781 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9841 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9901 ccagttcgat gtaacccact cgtgcaccca
//
