LOCUS       sequence-415786-        9807 bp    DNA     circular SYN 14-APR-2025
DEFINITION  A variant CE1 construct with B103 DNA polymerase, expressed from CMV
            or T7 promoters. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9807)
  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 9807)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: Polymerase variant click editor (CE1) -
            pCMV-T...
FEATURES             Location/Qualifiers
     source          1..9807
                     /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             6793..6813
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6822..6839
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6862..6879)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6868..7092
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7163..7179)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7163..7179)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7176..7198)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7187..7203
                     /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(7211..7217,7218..7235,7236..7241))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7256..7277
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7394..7411)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7565..8153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7645..7664)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8324..9115,9116..9184))
                     /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     8947..8966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9185..9289)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9368..9387)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(9559..9807,1..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 aagccctaga
     5041 aagatgttca gctgcgattt cgaaaccaca accaagctgg acgactgcag agtgtgggct
     5101 tacggctaca tggaaatcgg caacctggat aactacaaga ttggaaacag cctggatgag
     5161 ttcatgcagt gggtgatgga aatccaggcc gacctgtact tccacaacct gaagtttgac
     5221 ggcgccttca tagtgaactg gctggaaaga cacggcttca agtggagcaa cgagggcctg
     5281 cccaacacct acaataccat catttctaag atgggccagt ggtacatgat cgatatctgt
     5341 ttcggctata agggcaagcg gaagctgcac accgtgatct acgacagcct gaagaagctg
     5401 ccattccctg tcaaaaagat cgccaaagat tttcagctgc ctctgctgaa aggcgatatc
     5461 gactaccaca aggagcggcc tgtcggccac gaaatcacac cagaggaata tgaatacatc
     5521 aagaatgaca tcgagatcat cgcccgggcc ctggacatcc agttcaagca aggcctggat
     5581 agaatgaccg ctggctctga ttctctgaag gggttcaaag atatcctgag caccaagaag
     5641 ttcaacaagg tgttccccaa gttaagcctg cctatggaca aggaaatccg ctacgcctat
     5701 aggggcggct ttacatggct gaacgacaaa tacaaggaaa aggagattgg agaaggcatg
     5761 gtgttcgatg tgaactctct gtatccttcc cagatgtaca gcagacctct gccttacggc
     5821 gcccccatcg tgttccaggg caagtacgag aaggacgagc agtaccccct gtacatccaa
     5881 agaatcagat tcgagttcga gctgaaagag ggctatatcc ccaccatcca gatcaaaaaa
     5941 aaccccttct tcaagggcaa tgagtacctg aagaactccg gcggagagcc tgttgaactg
     6001 tacctgacca acgtggatct ggaactgatc caggagcact acgagctgta caacgtggag
     6061 tacattgacg gctttaaatt tagagaaaaa accggcttgt ttaaggactt catcgacaag
     6121 tggacctacg tgaagacaca cgaggaagga gcaaagaaac agctggccaa gctgatgctg
     6181 aacagcctgt acggcaagtt tgccagcaat cctgacgtga ccggcaaagt tccatacctc
     6241 aaagaggacg gatctctggg cttccgggtg ggcgacgaag aatacaagga ccccgtgtac
     6301 acccctatgg gagtgtttat cacagcctgg gccagattca ccaccatcac cgccgcccag
     6361 gcttgttacg accggatcat ctactgcgac acagacagca tccacctgac aggcaccgag
     6421 gtgcctgaga tcatcaagga catcgtggac cctaagaagc tgggctactg ggctcatgag
     6481 tccaccttca aaagagccaa gtacctgaga cagaagacct acatccagga catctacatg
     6541 aaggaagttg acggaaagct ggtggagtgc agccctgatg aggccaccac aaccaagttc
     6601 tccgtgaagt gcgccggcat gacagatacc atcaaaaaga aggtgacatt cgacaacttc
     6661 aaggtgggct tcagcagcaa gggaaagcct aaaccagtgc aagtgaacgg cggagtggtg
     6721 ctggtagaca gcgtgttcac aatcaagtct ggcggctcaa aaagaaccgc cgacggcagc
     6781 gaattcgagt ctcccaagaa gaagaggaaa gtctgaccgg tcatcatcac catcaccatt
     6841 gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     6901 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     6961 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     7021 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     7081 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgataccg tcgacctcta
     7141 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7201 caattccaca caacatacga gccggaagca taaagtgtaa agcctagggt gcctaatgag
     7261 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7321 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7381 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7441 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7501 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7561 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7621 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7681 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7741 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7801 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7861 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7921 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7981 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     8041 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8101 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8161 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8221 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8281 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8341 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8401 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8461 cgcgcgaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8521 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8581 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8641 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8701 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8761 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8821 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8881 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8941 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     9001 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     9061 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     9121 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9181 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9241 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9301 gaaaagtgcc acctgacgtc gacggatcgg gagatcgatc tcccgatccc ctagggtcga
     9361 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     9421 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     9481 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     9541 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     9601 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     9661 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     9721 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     9781 cccacttggc agtacatcaa gtgtatc
//
