LOCUS       sequence-275479-       11193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter expression plasmid for human codon optimized BE4max 
            C-to-T base editor with 
            xCas9(3.7)(D10A/A262T/R324L/S409I/E480K/E543D/M694I/E1219V) and 
            P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11193)
  AUTHORS   Walton RT, Christie KA, Whittaker MN, Kleinstiver BP
  TITLE     Unconstrained genome targeting with near-PAMless engineered 
            CRISPR-Cas9 variants.
  JOURNAL   Science. 2020 Apr 17;368(6488):290-296. doi: 
            10.1126/science.aba8853. Epub 2020 Mar 26.
  PUBMED    32217751
REFERENCE   2  (bases 1 to 11193)
  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"; doi: 
            "10.1126/science.aba8853"; journalName: "Science"; date: 
            "2020-04-17- 17"; volume: "368"; issue: "6488"; pages: "290-296"
COMMENT     Sequence Label: pCAG-CBE4max-xCas9(3.7)-P2A-EGFP (RTW4637)
FEATURES             Location/Qualifiers
     source          1..11193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     509..531
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     593..612
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             720..740
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             741..1424
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             1521..5621
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 mutant with broad PAM
                     compatibility (Hu et al., 2018)"
                     /label=xCas9-3.7
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1521..5621
                     /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
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5652..5900
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5931..6179
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6222..6242
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6261..6317
                     /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             join(6318..6320,6321..6323,6324..7037)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6318..6320,6321..6323,6324..7034)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6624..6643)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6971..6992
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7124..7143)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7189..7244
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7243..7262)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7605..7621)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7605..7621)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7618..7640)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7629..7645
                     /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(7653..7659,7660..7677,7678..7683))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7698..7719
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7778..7973
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7824..7959
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7979..8113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8016..8035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8070..8089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8181..8198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8352..8940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8432..8451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9111..9902,9903..9971))
                     /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     9734..9753
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9972..10076)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        10107..10486
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          join(10762..11193,1..585)
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
ORIGIN
        1 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
       61 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
      121 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
      181 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
      241 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
      301 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
      361 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
      421 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
      481 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
      541 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
      601 ctggttattg tgctgtctca tcattttggc aaagaattct gcagtcgacg gtaccgcggg
      661 cccgggatcc accggtcgcc accatgaaac ggacagccga cggaagcgag ttcgagtcac
      721 caaagaagaa gcggaaagtc tcctcagaga ctgggcctgt cgccgtcgat ccaaccctgc
      781 gccgccggat tgaacctcac gagtttgaag tgttctttga cccccgggag ctgagaaagg
      841 agacatgcct gctgtacgag atcaactggg gaggcaggca ctccatctgg aggcacacct
      901 ctcagaacac aaataagcac gtggaggtga acttcatcga gaagtttacc acagagcggt
      961 acttctgccc caataccaga tgtagcatca catggtttct gagctggtcc ccttgcggag
     1021 agtgtagcag ggccatcacc gagttcctgt ccagatatcc acacgtgaca ctgtttatct
     1081 acatcgccag gctgtatcac cacgcagacc caaggaatag gcagggcctg cgcgatctga
     1141 tcagctccgg cgtgaccatc cagatcatga cagagcagga gtccggctac tgctggcgga
     1201 acttcgtgaa ttattctcct agcaacgagg cccactggcc taggtaccca cacctgtggg
     1261 tgcgcctgta cgtgctggag ctgtattgca tcatcctggg cctgccccct tgtctgaata
     1321 tcctgcggag aaagcagccc cagctgacct tctttacaat cgccctgcag tcttgtcact
     1381 atcagaggct gccaccccac atcctgtggg ccacaggcct gaagtctgga ggatctagcg
     1441 gaggatcctc tggcagcgag acaccaggaa caagcgagtc agcaacacca gagagcagtg
     1501 gcggcagcag cggcggcagc gacaagaagt acagcatcgg cctggccatc ggcaccaact
     1561 ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc
     1621 tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca
     1681 gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac
     1741 ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg
     1801 acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc
     1861 ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca
     1921 tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct
     1981 atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga
     2041 accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc
     2101 tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca
     2161 gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga
     2221 atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca
     2281 acttcgacct ggccgaggat accaaactgc agctgagcaa ggacacctac gacgacgacc
     2341 tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga
     2401 acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg
     2461 cccccctgag cgcctctatg atcaagctgt acgacgagca ccaccaggac ctgaccctgc
     2521 tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga
     2581 gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt
     2641 tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca
     2701 gagaggacct gctgcggaag cagcggacct tcgacaacgg catcatcccc caccagatcc
     2761 acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg
     2821 acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc
     2881 tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc
     2941 cctggaactt cgagaaggtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga
     3001 tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt
     3061 acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga
     3121 gaaagcccgc cttcctgagc ggcgaccaga aaaaggccat cgtggacctg ctgttcaaga
     3181 ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct
     3241 tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc
     3301 acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca
     3361 ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac
     3421 ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga
     3481 gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt
     3541 ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatcc
     3601 agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg
     3661 gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga
     3721 agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca
     3781 agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga
     3841 agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga
     3901 tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact
     3961 acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg
     4021 actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca
     4081 aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg
     4141 tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga
     4201 gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg
     4261 gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc
     4321 tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag
     4381 tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag
     4441 tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa
     4501 ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg
     4561 tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca
     4621 agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg
     4681 gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg
     4741 ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg
     4801 tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga
     4861 acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg
     4921 acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca
     4981 agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg
     5041 agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga
     5101 tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg
     5161 cctctgccgg cgtgctgcag aagggaaacg aactggccct gccctccaaa tatgtgaact
     5221 tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga
     5281 aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg
     5341 agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca
     5401 acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc
     5461 tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga
     5521 ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc
     5581 tgtacgagac acggatcgac ctgtctcagc tgggaggtga cagcggcggg agcggcggga
     5641 gcggggggag cactaatctg agcgacatca ttgagaagga gactgggaaa cagctggtca
     5701 ttcaggagtc catcctgatg ctgcctgagg aggtggagga agtgatcggc aacaagccag
     5761 agtctgacat cctggtgcac accgcctacg acgagtccac agatgagaat gtgatgctgc
     5821 tgacctctga cgcccccgag tataagcctt gggccctggt catccaggat tctaacggcg
     5881 agaataagat caagatgctg agcggaggat ccggaggatc tggaggcagc accaacctgt
     5941 ctgacatcat cgagaaggag acaggcaagc agctggtcat ccaggagagc atcctgatgc
     6001 tgcccgaaga agtcgaagaa gtgatcggaa acaagcctga gagcgatatc ctggtccata
     6061 ccgcctacga cgagagtacc gacgaaaatg tgatgctgct gacatccgac gccccagagt
     6121 ataagccctg ggctctggtc atccaggatt ccaacggaga gaacaaaatc aaaatgctgt
     6181 ctggcggctc aaaaagaacc gccgacggca gcgaattcga gcccaagaag aagaggaaag
     6241 tcggaggcgg tggaagcggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg
     6301 aagagaatcc tggaccgatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     6361 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     6421 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     6481 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     6541 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     6601 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     6661 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     6721 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     6781 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     6841 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     6901 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     6961 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     7021 acgagctgta caagtgaagc ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg
     7081 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     7141 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     7201 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     7261 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     7321 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     7381 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     7441 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     7501 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     7561 tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     7621 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7681 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7741 gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca
     7801 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     7861 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     7921 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa
     7981 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     8041 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     8101 tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     8161 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     8221 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     8281 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     8341 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     8401 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8461 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     8521 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8581 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8641 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8701 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     8761 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     8821 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8881 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8941 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     9001 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     9061 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     9121 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     9181 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     9241 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     9301 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     9361 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     9421 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     9481 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     9541 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9601 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9661 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     9721 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     9781 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     9841 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     9901 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9961 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    10021 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    10081 tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagttatt aatagtaatc
    10141 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
    10201 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
    10261 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
    10321 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
    10381 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
    10441 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
    10501 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
    10561 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggsgggc gcgcgscggg
    10621 gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag
    10681 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
    10741 aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc
    10801 gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc
    10861 gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg acggcttgtt
    10921 tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt gtgcgggggg
    10981 agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg
    11041 ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg
    11101 cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac
    11161 aaaggctgcg tgcggggtgt gtgcgtgggg ggg
//
