LOCUS       sequence-320189-        8403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian CG-to-GC base editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8403)
  AUTHORS   Koblan LW, Arbab M, Shen MW, Hussmann JA, Anzalone AV, Doman JL, 
            Newby GA, Yang D, Mok B, Replogle JM, Xu A, Sisley TA, Weissman JS, 
            Adamson B, Liu DR
  TITLE     Efficient C*G-to-G*C base editors developed using CRISPRi screens, 
            target-library analysis, and machine learning.
  JOURNAL   Nat Biotechnol. 2021 Jun 28. pii: 10.1038/s41587-021-00938-z. doi: 
            10.1038/s41587-021-00938-z.
  PUBMED    34183861
REFERENCE   2  (bases 1 to 8403)
  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. 2021 Jun 28. pii: 10.1038/s41587-021-00938-z. doi: 
            10.1038/s41587-021-00938-z."
COMMENT     Sequence Label: pCMV_Anc689-nCas9
FEATURES             Location/Qualifiers
     source          1..8403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     246..265
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        246..264
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             314..334
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1115..1138,1139..1141,1142..5215)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1115..5215
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5258..5278
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5287..5304
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5327..5344)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5333..5557
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5628..5644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5628..5644)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5641..5663)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5652..5668
                     /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(5676..5682,5683..5700,5701..5706))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5721..5742
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5859..5876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6030..6618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6110..6129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6789..7580,7581..7649))
                     /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     7412..7431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7650..7754)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7833..7852)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8024..8403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gagatccgcg
      241 gccgctaata cgactcacta tagggagagc cgccaccatg aaacggacag ccgacggaag
      301 cgagttcgag tcaccaaaga agaagcggaa agtctcctca gagactgggc ctgtcgcagt
      361 ggacccaacc ctgaggagac ggattgagcc ccatgaattt gaagtgttct ttgacccaag
      421 ggagctgagg aaggagacat gcctgctgta cgagatcaag tggggcacaa gccacaagat
      481 ctggcgccac agctccaaga acaccacaaa gcacgtggaa gtgaatttca tcgagaagtt
      541 tacctccgag cggcacttct gcccctctac cagctgttcc atcacatggt ttctgtcttg
      601 gagcccttgc ggcgagtgtt ccaaggccat caccgagttc ctgtctcagc accctaacgt
      661 gaccctggtc atctacgtgg cccggctgta tcaccacatg gaccagcaga acaggcaggg
      721 cctgcgcgat ctggtgaatt ctggcgtgac catccagatc atgacagccc cagagtacga
      781 ctattgctgg cggaacttcg tgaattatcc acctggcaag gaggcacact ggccaagata
      841 cccacccctg tggatgaagc tgtatgcact ggagctgcac gcaggaatcc tgggcctgcc
      901 tccatgtctg aatatcctgc ggagaaagca gccccagctg acatttttca ccattgctct
      961 gcagtcttgt cactatcagc ggctgcctcc tcatattctg tgggccacag gcctgaagtc
     1021 tggaggatct agcggaggat cctctggcag cgagacacca ggaacaagcg agtcagcaac
     1081 accagagagc agtggcggca gcagcggcgg cagcgacaag aagtacagca tcggcctggc
     1141 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     1201 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     1261 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     1321 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     1381 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     1441 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     1501 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     1561 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     1621 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     1681 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     1741 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     1801 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     1861 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     1921 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     1981 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     2041 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     2101 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     2161 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     2221 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     2281 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     2341 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     2401 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     2461 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     2521 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     2581 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     2641 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     2701 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     2761 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     2821 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     2881 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     2941 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     3001 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     3061 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     3121 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     3181 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     3241 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     3301 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     3361 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     3421 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     3481 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     3541 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     3601 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     3661 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     3721 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     3781 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     3841 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     3901 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     3961 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     4021 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     4081 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     4141 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     4201 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     4261 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     4321 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     4381 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     4441 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     4501 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     4561 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     4621 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     4681 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     4741 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     4801 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     4861 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     4921 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     4981 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     5041 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     5101 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     5161 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg
     5221 cggctcaaaa agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcta
     5281 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
     5341 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     5401 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     5461 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     5521 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     5581 tggggctcga taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5641 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5701 tgtaaagcct agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5761 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5821 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5881 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5941 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6001 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6061 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6121 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6181 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6241 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6301 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6361 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6421 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6481 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6541 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6601 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg
     6661 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6721 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6781 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6841 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6901 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6961 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7021 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7081 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7141 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7201 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7261 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7321 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7381 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7441 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7501 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7561 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7621 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7681 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7741 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7801 cgatctcccg atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     7861 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7921 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     7981 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     8041 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     8101 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     8161 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     8221 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     8281 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     8341 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     8401 atg
//
