LOCUS       sequence-447694-        9591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for TadCBEd using SpCas9 CBE 
            enzyme with KWRQLC amino acids at positions 
            1135,1136,1218,1219,1335,T1337 with C-term BPNLS-P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9591)
  AUTHORS   Silverstein RA, Kim N, Kroell AS, Walton RT, Delano J, Butcher RM, 
            Pacesa M, Smith BK, Christie KA, Ha LL, Meis RJ, Clark AB, Spinner 
            AD, Lazzarotto CR, Li Y, Matsubara A, Urbina EO, Dahl GA, Correia 
            BE, Marks DS, Tsai SQ, Pinello L, De Ravin SS, Liu Q, Kleinstiver BP
  TITLE     Custom CRISPR-Cas9 PAM variants via scalable engineering and machine
            learning.
  JOURNAL   Nature. 2025 Apr 22. doi: 10.1038/s41586-025-09021-y.
  PUBMED    40262634
REFERENCE   2  (bases 1 to 9591)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2025 Apr 22. doi: 10.1038/s41586-025-09021-y."
COMMENT     Sequence Label: pCMV-T7-TadCBEd-SpCas9(KWRQLC)-P2A-EGFP (BKS1026)
FEATURES             Location/Qualifiers
     source          1..9591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        116..495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        496..699
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     650..670
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     741..760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        741..759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             809..829
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1424..5524
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFKWPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARQLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKLYCSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5555..5803
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5834..6082
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6125..6145
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6155..6211
                     /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(6212..6214,6215..6217,6218..6928)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6212..6214,6215..6217,6218..6928)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6518..6537)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6865..6886
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6941..6961
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6970..6987
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7010..7027)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7016..7240
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7311..7327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7311..7327)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7324..7346)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7335..7351
                     /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(7359..7365,7366..7383,7384..7389))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7404..7425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7542..7559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7713..8301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7793..7812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8472..9263,9264..9332))
                     /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     9095..9114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9333..9437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9516..9535)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
       61 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      121 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      361 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      421 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      481 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      541 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      601 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      661 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      721 cgctagagat ccgcggccgc taatacgact cactataggg agagccgcca ccatgaaacg
      781 gacagccgac ggaagcgagt tcgagtcacc aaagaagaag cggaaagtct ctgaggtgga
      841 gttttcccac gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgcgatga
      901 gaggaaggcc cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg
      961 gaacagagcc atcggcctgc acgacccaac agcccatgcc gaaattatcg ccctgagaca
     1021 gggcggcctg gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga
     1081 gccttgcgtg atgtgcgccg gcgccatgat caactctagg atcggccgcg tggtgtttgg
     1141 cgtgaggaac agcaaaagag gcgccgcagg ctccctgatg aacgtgctga actaccccgg
     1201 catgaatcac cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct
     1261 gtgcgatttc taccggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc
     1321 catcaactcc ggaggatcta gcggaggctc ctctggctct gagacacctg gcacaagcga
     1381 gagcgcaaca cctgaaagca gcgggggcag cagcgggggg tcagacaaga agtacagcat
     1441 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
     1501 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
     1561 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
     1621 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
     1681 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
     1741 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
     1801 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
     1861 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
     1921 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
     1981 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
     2041 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
     2101 ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg
     2161 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
     2221 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
     2281 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     2341 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
     2401 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     2461 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
     2521 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     2581 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     2641 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
     2701 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     2761 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     2821 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
     2881 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     2941 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
     3001 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
     3061 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     3121 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     3181 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     3241 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     3301 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     3361 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
     3421 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     3481 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     3541 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     3601 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     3661 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     3721 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     3781 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     3841 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     3901 actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct
     3961 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag
     4021 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
     4081 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
     4141 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     4201 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
     4261 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     4321 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     4381 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     4441 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     4501 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     4561 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     4621 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
     4681 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     4741 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
     4801 ccctaagaag tacggcggct tcaagtggcc caccgtggcc tattctgtgc tggtggtggc
     4861 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
     4921 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     4981 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     5041 aaacggccgg aagagaatgc tggcctctgc cagacagctg cagaagggaa acgaactggc
     5101 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     5161 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
     5221 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     5281 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
     5341 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
     5401 tgacaccacc atcgaccgga agctgtactg cagcaccaaa gaggtgctgg acgccaccct
     5461 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     5521 tgacagcggc gggagcggcg ggagcggggg gagcactaat ctgagcgaca tcattgagaa
     5581 ggagactggg aaacagctgg tcattcagga gtccatcctg atgctgcctg aggaggtgga
     5641 ggaagtgatc ggcaacaagc cagagtctga catcctggtg cacaccgcct acgacgagtc
     5701 cacagatgag aatgtgatgc tgctgacctc tgacgccccc gagtataagc cttgggccct
     5761 ggtcatccag gattctaacg gcgagaataa gatcaagatg ctgagcggag gatccggagg
     5821 atctggaggc agcaccaacc tgtctgacat catcgagaag gagacaggca agcagctggt
     5881 catccaggag agcatcctga tgctgcccga agaagtcgaa gaagtgatcg gaaacaagcc
     5941 tgagagcgat atcctggtcc ataccgccta cgacgagagt accgacgaaa atgtgatgct
     6001 gctgacatcc gacgccccag agtataagcc ctgggctctg gtcatccagg attccaacgg
     6061 agagaacaaa atcaaaatgc tgtctggcgg ctcaaaaaga accgccgacg gcagcgaatt
     6121 cgagcccaag aagaagagga aagtcggaag cggagctact aacttcagcc tgctgaagca
     6181 ggctggagac gtggaggaga accctggacc tatggtgagc aagggcgagg agctgttcac
     6241 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     6301 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     6361 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct atggagtgca
     6421 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     6481 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     6541 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     6601 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     6661 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     6721 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     6781 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     6841 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     6901 cactctcggc atggacgagc tgtacaagtc tggtggttct cccaagaaga agaggaaagt
     6961 ctaaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg
     7021 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     7081 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     7141 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     7201 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     7261 agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     7321 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7381 aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7441 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7501 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7561 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7621 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7681 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7741 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7801 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7861 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7921 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7981 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     8041 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     8101 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     8161 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8221 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     8281 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacactcag
     8341 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8401 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8461 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8521 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8581 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8641 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8701 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8761 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8821 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8881 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8941 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     9001 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     9061 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     9121 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     9181 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     9241 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9301 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9361 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9421 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     9481 gatcgatctc ccgatcccct agggtcgact ctcagtacaa tctgctctga tgccgcatag
     9541 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg c
//
