LOCUS       sequence-379830-        8745 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of CABE-T3.1 base editor in mammalian cells, CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8745)
  AUTHORS   Lam DK, Feliciano PR, Arif A, Bohnuud T, Fernandez TP, Gehrke JM, 
            Grayson P, Lee KD, Ortega MA, Sawyer C, Schwaegerle ND, Peraro L, 
            Young L, Lee SJ, Ciaramella G, Gaudelli NM
  TITLE     Improved cytosine base editors generated from TadA variants.
  JOURNAL   Nat Biotechnol. 2023 Jan 9. doi: 10.1038/s41587-022-01611-9.
  PUBMED    36624149
REFERENCE   2  (bases 1 to 8745)
  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. 2023 Jan 9. doi: 10.1038/s41587-022-01611-9."
COMMENT     Sequence Label: CABE-T3.1
FEATURES             Location/Qualifiers
     source          1..8745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        103..482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        483..686
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     637..657
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     728..747
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        728..746
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             763..1260
                     /codon_start=1
                     /product="DNA adenosine deaminase evolved from the E. coli 
                     tRNA adenosine deaminase TadA (Gaudelli et al., 2017)"
                     /label=TadA*(7.10)
                     /translation="SEVEFSHEYWMRHALTLAKRARDERSVPVGAVLVLNNRVIGEGWN
                     RAKGLHDPTAHAEIMALRQGGLVMQNYRLYDATLYTTFEPCVMCAGAMIHSRIGRVVFG
                     VRNAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLCRFFRMPRRVFNAQKKAQS
                     STD"
     CDS             join(1357..1380,1381..1383,1384..5457)
                     /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             1357..5457
                     /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             5488..5736
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5767..6015
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6016..6036
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6082..6102
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6111..6128
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6151..6168)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6157..6381
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6452..6468)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6452..6468)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6465..6487)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6476..6492
                     /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(6500..6506,6507..6524,6525..6530))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6545..6566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6683..6700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6854..7442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6934..6953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7613..8404,8405..8473))
                     /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     8236..8255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8474..8578)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8657..8676)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
       61 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      121 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      181 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      241 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      301 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      361 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      421 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      481 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      541 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      601 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      661 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg
      721 cggccgctaa tacgactcac tatagggaga gccgccacca tgtccgaagt cgagttttcc
      781 catgagtact ggatgagaca cgcattgact ctcgcaaaga gggctcgaga tgaacgcagc
      841 gtgcccgtgg gggcagtact cgtgctcaac aatcgcgtaa tcggcgaagg ttggaatagg
      901 gcaaagggac tccacgaccc cactgcacat gcggaaatca tggcccttcg acagggaggg
      961 cttgtgatgc agaattatcg actttatgat gcgacgctgt acaccacgtt tgaaccttgc
     1021 gtaatgtgcg cgggagctat gattcactcc cgcattggac gagttgtatt cggtgttcgc
     1081 aacgccaaga cgggtgccgc aggttcactg atggacgtgc tgcatcatcc aggcatgaac
     1141 caccgggtag aaatcacaga aggcatattg gcggacgaat gtgcggcgct gttgtgtcgt
     1201 ttttttcgca tgcccaggcg ggtctttaac gcccagaaaa aagcacaatc ctctactgac
     1261 tctggtggtt cttctggtgg ttctagcggc agcgagactc ccgggacctc agagtccgcc
     1321 acacccgaaa gttctggtgg ttcttctggt ggttctgaca agaagtacag catcggcctg
     1381 gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     1441 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     1501 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
     1561 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     1621 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     1681 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     1741 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     1801 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     1861 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     1921 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     1981 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     2041 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
     2101 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     2161 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     2221 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     2281 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     2341 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     2401 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     2461 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     2521 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     2581 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     2641 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     2701 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     2761 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     2821 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     2881 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     2941 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     3001 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     3061 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     3121 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     3181 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     3241 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     3301 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     3361 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     3421 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     3481 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     3541 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     3601 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     3661 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     3721 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     3781 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     3841 atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac
     3901 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
     3961 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
     4021 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     4081 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     4141 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
     4201 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     4261 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     4321 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     4381 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     4441 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     4501 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
     4561 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
     4621 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     4681 atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
     4741 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
     4801 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
     4861 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
     4921 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     4981 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
     5041 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     5101 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
     5161 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
     5221 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
     5281 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
     5341 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
     5401 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggtgactct
     5461 ggtggaagcg gaggatctgg cggcagcacc aatctgagcg acatcatcga gaaagagaca
     5521 ggcaagcagc tggtcatcca agagtccatc ctgatgctgc ctgaagaggt ggaagaagtg
     5581 atcggcaaca agcccgagtc cgacatcctg gtgcacaccg cctacgatga gagcaccgac
     5641 gagaacgtga tgctgctgac ctctgacgcc cctgagtaca agccttgggc tctcgtgatc
     5701 caggacagca acggcgagaa caagatcaag atgctgagcg gcggctctgg tggctctggc
     5761 ggatctacaa acctgtccga tattattgag aaagaaaccg ggaaacagct cgtgattcaa
     5821 gagtctattc tcatgctccc ggaagaagtc gaggaagtca ttggaaacaa gcctgagagc
     5881 gatattctgg tccatacagc ctacgacgag tctaccgatg agaatgtcat gctcctcacc
     5941 agcgacgctc ccgagtataa gccatgggca cttgtcattc aggactccaa tggggaaaac
     6001 aaaatcaaaa tgctcccaaa gaaaaaacgc aaggtggagg gagctgataa gcgcaccgcc
     6061 gatggttccg agttcgaaag ccccaagaag aagaggaaag tctaaccggt catcatcacc
     6121 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     6181 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     6241 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     6301 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     6361 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgataccgt
     6421 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6481 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctagggtg
     6541 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6601 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6661 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6721 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6781 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6841 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6901 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6961 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7021 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7081 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7141 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7201 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7261 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7321 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7381 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7441 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7501 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7561 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7621 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7681 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7741 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7801 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7861 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7921 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7981 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8041 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8101 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8161 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8221 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8281 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8341 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8401 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8461 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8521 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8581 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc
     8641 tagggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     8701 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aattt
//
