LOCUS       sequence-424802-        8778 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express CBE6b (with SpCas9) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8778)
  AUTHORS   Zhang E, Neugebauer ME, Krasnow NA, Liu DR
  TITLE     Phage-assisted evolution of highly active cytosine base editors with
            enhanced selectivity and minimal sequence context preference.
  JOURNAL   Nat Commun. 2024 Feb 24;15(1):1697. doi: 10.1038/s41467-024-45969-7.
  PUBMED    38402281
REFERENCE   2  (bases 1 to 8778)
  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 
            Commun."; date: "2024-02-24"; volume: "15(1):1697. doi"; pages: " 
            10.1038/s41467-024-45969-7"
COMMENT     Sequence Label: SpCas9-CBE6b
FEATURES             Location/Qualifiers
     source          1..8778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     610..629
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        610..628
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             678..698
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1296..1319,1320..1322,1323..5396)
                     /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             1296..5396
                     /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             5427..5675
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5706..5954
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5997..6017
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6026..6043
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6066..6083)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6072..6296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6367..6383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6367..6383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6380..6402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6391..6407
                     /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(6415..6421,6422..6439,6440..6445))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6460..6481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6598..6615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6769..7357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6849..6868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7528..8319,8320..8388))
                     /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     8151..8170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8389..8493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8572..8591)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8763..8778,1..364)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc
      601 cgcggccgct aatacgactc actataggga gagccgccac catgaaacgg acagccgacg
      661 gaagcgagtt cgagtcacca aagaagaagc ggaaagtcag ttctgaggtg gagttttccc
      721 acgagtactg gatgagacat gccctgaccc tggccaagag ggcacgggat gagggagagg
      781 cgcctgtggg agccgtgctg gtgctgaaca atagagtgat cggcgagggc tgggtgagac
      841 gtatcggcct gcacgaccca acagcccatg ccgaaattat ggccctgaga cagggcggcc
      901 tggtcatgca gaaccccaga ctgattgacg ccaccctgta cgtgacattc gagccttgcg
      961 tgatgtgcgc cggcgccatg atcaactcta ggatcggccg cgtggtgttt ggcgtgagga
     1021 actcaaaaag aggcgccgca ggctccctga tgaacgtgct gaactacccc ggcatgaatc
     1081 accgcgtcga aattaccgag ggaatcctgg cagatgaatg tgccgccctg ctgtgcgatt
     1141 tctatcggat gcctagacag gtgttcaatg ctcagaagaa ggcccagagc tccatcaact
     1201 ctggcggatc tagcggagga tcctctggca gcgagacacc aggaacaagc gagtcagcaa
     1261 caccagagag cagtggcggc agcagcggcg gcagcgacaa gaagtacagc atcggcctgg
     1321 ccatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     1381 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     1441 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     1501 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     1561 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     1621 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     1681 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     1741 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     1801 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     1861 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     1921 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     1981 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     2041 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     2101 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     2161 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     2221 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     2281 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     2341 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     2401 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     2461 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     2521 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     2581 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     2641 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     2701 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     2761 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     2821 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     2881 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     2941 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     3001 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     3061 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     3121 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     3181 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     3241 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     3301 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     3361 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     3421 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     3481 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     3541 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     3601 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     3661 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     3721 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     3781 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
     3841 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     3901 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     3961 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     4021 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     4081 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     4141 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     4201 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     4261 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     4321 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     4381 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     4441 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     4501 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     4561 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     4621 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     4681 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     4741 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     4801 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     4861 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     4921 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     4981 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     5041 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     5101 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     5161 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     5221 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     5281 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     5341 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggtgacagcg
     5401 gcgggagcgg cgggagcggg gggagcacta atctgagcga catcattgag aaggagactg
     5461 ggaaacagct ggtcattcag gagtccatcc tgatgctgcc tgaggaggtg gaggaagtga
     5521 tcggcaacaa gccagagtct gacatcctgg tgcacaccgc ctacgacgag tccacagatg
     5581 agaatgtgat gctgctgacc tctgacgccc ccgagtataa gccttgggcc ctggtcatcc
     5641 aggattctaa cggcgagaat aagatcaaga tgctgagcgg aggatccgga ggatctggag
     5701 gcagcaccaa cctgtctgac atcatcgaga aggagacagg caagcagctg gtcatccagg
     5761 agagcatcct gatgctgccc gaagaagtcg aagaagtgat cggaaacaag cctgagagcg
     5821 atatcctggt ccataccgcc tacgacgaga gtaccgacga aaatgtgatg ctgctgacat
     5881 ccgacgcccc agagtataag ccctgggctc tggtcatcca ggattccaac ggagagaaca
     5941 aaatcaaaat gctgtctggc ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca
     6001 agaagaagag gaaagtctaa ccggtcatca tcaccatcac cattgagttt aaacccgctg
     6061 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     6121 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     6181 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     6241 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     6301 tgaggcggaa agaaccagct ggggctcgat accgtcgacc tctagctaga gcttggcgta
     6361 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6421 acgagccgga agcataaagt gtaaagccta gggtgcctaa tgagtgagct aactcacatt
     6481 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6541 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgcactt ccgcttcctc
     6601 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6661 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6721 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6781 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6841 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6901 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6961 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7021 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7081 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7141 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7201 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7261 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7321 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7381 ggggtctgac actcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7441 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7501 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7561 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7621 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc
     7681 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7741 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7801 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7861 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7921 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7981 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8041 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8101 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8161 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8221 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8281 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8341 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8401 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8461 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8521 cgtcgacgga tcgggagatc gatctcccga tcccctaggg tcgactctca gtacaatctg
     8581 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     8641 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     8701 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     8761 ttgacattga ttattgac
//
