LOCUS       sequence-447695-        9591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for TadCBEd using SpCas9 CBE 
            enzyme with LWKYQS 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(LWKYQS)-P2A-EGFP (BKS1014)
FEATURES             Location/Qualifiers
     source          1..9591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        94..473
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        474..677
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     628..648
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     719..738
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        719..737
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             787..807
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1402..5502
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFLWPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKYLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYSSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5533..5781
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5812..6060
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6103..6123
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6133..6189
                     /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(6190..6192,6193..6195,6196..6906)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6190..6192,6193..6195,6196..6906)
                     /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(6496..6515)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6843..6864
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6919..6939
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6948..6965
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6988..7005)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6994..7218
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7289..7305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7289..7305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7302..7324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7313..7329
                     /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(7337..7343,7344..7361,7362..7367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7382..7403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7520..7537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7691..8279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7771..7790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8450..9241,9242..9310))
                     /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     9073..9092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9311..9415)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9494..9513)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
       61 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
      121 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      181 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      241 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      301 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      361 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      421 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      481 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      541 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      601 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      661 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagagatcc gcggccgcta
      721 atacgactca ctatagggag agccgccacc atgaaacgga cagccgacgg aagcgagttc
      781 gagtcaccaa agaagaagcg gaaagtctct gaggtggagt tttcccacga gtactggatg
      841 agacatgccc tgaccctggc caagagggca cgcgatgaga ggaaggcccc tgtgggagcc
      901 gtgctggtgc tgaacaatag agtgatcggc gagggctgga acagagccat cggcctgcac
      961 gacccaacag cccatgccga aattatcgcc ctgagacagg gcggcctggt catgcagaac
     1021 tacagactga ttgacgccac cctgtacgtg acattcgagc cttgcgtgat gtgcgccggc
     1081 gccatgatca actctaggat cggccgcgtg gtgtttggcg tgaggaacag caaaagaggc
     1141 gccgcaggct ccctgatgaa cgtgctgaac taccccggca tgaatcaccg cgtcgaaatt
     1201 accgagggaa tcctggcaga tgaatgtgcc gccctgctgt gcgatttcta ccggatgcct
     1261 agacaggtgt tcaatgctca gaagaaggcc cagagctcca tcaactccgg aggatctagc
     1321 ggaggctcct ctggctctga gacacctggc acaagcgaga gcgcaacacc tgaaagcagc
     1381 gggggcagca gcggggggtc agacaagaag tacagcatcg gcctggccat cggcaccaac
     1441 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     1501 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     1561 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     1621 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     1681 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     1741 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     1801 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     1861 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     1921 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     1981 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     2041 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     2101 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     2161 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     2221 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     2281 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     2341 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     2401 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     2461 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     2521 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     2581 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     2641 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     2701 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     2761 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     2821 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     2881 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     2941 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     3001 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     3061 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     3121 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     3181 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     3241 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     3301 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     3361 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     3421 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     3481 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     3541 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     3601 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     3661 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     3721 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     3781 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     3841 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     3901 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     3961 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     4021 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     4081 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     4141 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     4201 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     4261 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     4321 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     4381 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     4441 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     4501 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     4561 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     4621 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     4681 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     4741 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     4801 ctgtggccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     4861 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     4921 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     4981 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     5041 gcctctgcca agtacctgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     5101 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     5161 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     5221 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     5281 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     5341 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     5401 cagtacagca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     5461 ctgtacgaga cacggatcga cctgtctcag ctgggaggtg acagcggcgg gagcggcggg
     5521 agcgggggga gcactaatct gagcgacatc attgagaagg agactgggaa acagctggtc
     5581 attcaggagt ccatcctgat gctgcctgag gaggtggagg aagtgatcgg caacaagcca
     5641 gagtctgaca tcctggtgca caccgcctac gacgagtcca cagatgagaa tgtgatgctg
     5701 ctgacctctg acgcccccga gtataagcct tgggccctgg tcatccagga ttctaacggc
     5761 gagaataaga tcaagatgct gagcggagga tccggaggat ctggaggcag caccaacctg
     5821 tctgacatca tcgagaagga gacaggcaag cagctggtca tccaggagag catcctgatg
     5881 ctgcccgaag aagtcgaaga agtgatcgga aacaagcctg agagcgatat cctggtccat
     5941 accgcctacg acgagagtac cgacgaaaat gtgatgctgc tgacatccga cgccccagag
     6001 tataagccct gggctctggt catccaggat tccaacggag agaacaaaat caaaatgctg
     6061 tctggcggct caaaaagaac cgccgacggc agcgaattcg agcccaagaa gaagaggaaa
     6121 gtcggaagcg gagctactaa cttcagcctg ctgaagcagg ctggagacgt ggaggagaac
     6181 cctggaccta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     6241 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     6301 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     6361 tggcccaccc tcgtgaccac cctgacctat ggagtgcagt gcttcagccg ctaccccgac
     6421 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     6481 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     6541 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     6601 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     6661 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     6721 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     6781 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     6841 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     6901 tacaagtctg gtggttctcc caagaagaag aggaaagtct aaccggtcat catcaccatc
     6961 accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     7021 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     7081 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     7141 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     7201 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga
     7261 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     7321 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct
     7381 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7441 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7501 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7561 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7621 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7681 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7741 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7801 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7861 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7921 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7981 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     8041 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     8101 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     8161 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8221 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8281 aagatccttt gatcttttct acggggtctg acactcagtg gaacgaaaac tcacgttaag
     8341 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8401 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8461 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8521 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8581 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8641 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8701 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8761 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8821 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8881 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8941 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     9001 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     9061 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9121 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9181 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9241 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9301 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9361 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9421 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcgatctccc gatcccctag
     9481 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     9541 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca a
//
