LOCUS       sequence-460275-        9582 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express N2-BE4 base editor in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9582)
  TITLE     Evolved cytidine and adenine base editors with high precision and 
            minimized off-target activity by a continuous directed evolution 
            system in mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9582)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCMV-N2-BE4-puro
FEATURES             Location/Qualifiers
     source          1..9582
                     /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             445..465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1198..5298
                     /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
                     QLPEKYKEIFFDQSKNGYAGYINGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5329..5577
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5608..5856
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5899..5919
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5935..5991
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5992..6591
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5992..6011)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6488..6508
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             6597..6614
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6637..6654)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6643..6867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6938..6954)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6938..6954)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6951..6973)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6962..6978
                     /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(6986..6992,6993..7010,7011..7016))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7031..7052
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7169..7186)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7340..7928)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7420..7439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8099..8890,8891..8959))
                     /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     8722..8741
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8960..9064)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9143..9162)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(9334..9582,1..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 gaaacggaca
      421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctcctc agagactggg
      481 cctgtcgccg tcgatccaac cctgcgccgc cggattgaac ctcacgagtt tgaagtgttc
      541 tttgaccccc gggagctccg caaggagacc tgcctgcttt acgaaattaa ttgggggggc
      601 cggcactcca tttggcgaca tacatcacag aacactaaca agcacgtcga agtcaacttc
      661 atcgagaagt tcacgacaga aagatatttc tgtccgaaca caaggtgcag cattacctgg
      721 tttctcagct ggagcccatg cggcgaatgt agtagggcca tcactgaatt cctgtcaagg
      781 tatccccacg tcactctgtt tatttacatc gcaaggctgt accaccacgc tgacccccgc
      841 aatcgacaag gcctgcggga tttgatctct tcaggtgtgg ctatccaaat tatgactggg
      901 caggagtcag gatactgctg gagaaacttt gtgaattata gcccgagtaa tgaagcccac
      961 tggcctaggt atccccatct gtgggtacga ccgtacgttc ttgaactgta ctgcatcata
     1021 ctgggcctgc ctccttgtct caacattctg agaaggaagc agccacagct gacattcttt
     1081 accatcgctc ttcagtcttg tcattaccag cgactgcccc cacacattct ctgggccacc
     1141 gggttgaaaa gcggcagcga gactcccggg acctcagagt ccgccacacc cgaaagtgac
     1201 aagaagtaca gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc
     1261 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
     1321 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
     1381 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
     1441 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
     1501 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
     1561 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
     1621 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
     1681 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     1741 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     1801 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     1861 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     1921 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     1981 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     2041 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     2101 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     2161 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     2221 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     2281 attaacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     2341 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     2401 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     2461 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     2521 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     2581 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     2641 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     2701 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     2761 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     2821 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     2881 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     2941 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     3001 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     3061 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     3121 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     3181 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     3241 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     3301 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     3361 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     3421 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     3481 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     3541 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     3601 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     3661 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     3721 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     3781 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     3841 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     3901 gccgaaagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     3961 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     4021 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     4081 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     4141 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     4201 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     4261 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     4321 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     4381 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     4441 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     4501 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     4561 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     4621 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     4681 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     4741 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     4801 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     4861 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     4921 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     4981 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     5041 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     5101 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     5161 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     5221 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     5281 tctcagctgg gaggtgacag cggcgggagc ggcgggagcg gggggagcac taatctgagc
     5341 gacatcattg agaaggagac tgggaaacag ctggtcattc aggagtccat cctgatgctg
     5401 cctgaggagg tggaggaagt gatcggcaac aagccagagt ctgacatcct ggtgcacacc
     5461 gcctacgacg agtccacaga tgagaatgtg atgctgctga cctctgacgc ccccgagtat
     5521 aagccttggg ccctggtcat ccaggattct aacggcgaga ataagatcaa gatgctgagc
     5581 ggaggatccg gaggatctgg aggcagcacc aacctgtctg acatcatcga gaaggagaca
     5641 ggcaagcagc tggtcatcca ggagagcatc ctgatgctgc ccgaagaagt cgaagaagtg
     5701 atcggaaaca agcctgagag cgatatcctg gtccataccg cctacgacga gagtaccgac
     5761 gaaaatgtga tgctgctgac atccgacgcc ccagagtata agccctgggc tctggtcatc
     5821 caggattcca acggagagaa caaaatcaaa atgctgtctg gcggctcaaa aagaaccgcc
     5881 gacggcagcg aattctctcc caagaagaag aggaaagtcg ctagcggcag cggcgccacc
     5941 aacttcagcc tgctgaagca ggccggcgac gtggaggaga accccggccc catgaccgag
     6001 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     6061 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     6121 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     6181 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     6241 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     6301 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     6361 ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     6421 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     6481 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     6541 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accggtcatc
     6601 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     6661 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     6721 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     6781 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     6841 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     6901 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6961 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7021 agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7081 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7141 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7201 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7261 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7321 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7381 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7441 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7501 cctttttccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7561 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7621 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7681 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7741 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7801 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7861 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7921 gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg aacgaaaact
     7981 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8041 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8101 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8161 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8221 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8281 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8341 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8401 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8461 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8521 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8581 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8641 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8701 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8761 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8821 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8881 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8941 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9001 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9061 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
     9121 atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     9181 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     9241 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     9301 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     9361 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     9421 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     9481 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     9541 atttacggta aactgcccac ttggcagtac atcaagtgta tc
//
