LOCUS       sequence-447696-        9591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for TadCBEd using SpCas9 CBE 
            enzyme with LWRYEK 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(LWRYEK)-P2A-EGFP (BKS1022)
FEATURES             Location/Qualifiers
     source          1..9591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..498
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        499..702
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     653..673
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     744..763
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        744..762
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             812..832
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1427..5527
                     /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
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARYLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYKSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5558..5806
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5837..6085
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6128..6148
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6158..6214
                     /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(6215..6217,6218..6220,6221..6931)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6215..6217,6218..6220,6221..6931)
                     /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(6521..6540)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6868..6889
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6944..6964
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6973..6990
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7013..7030)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7019..7243
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7314..7330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7314..7330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7327..7349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7338..7354
                     /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(7362..7368,7369..7386,7387..7392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7407..7428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7545..7562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7716..8304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7796..7815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8475..9266,9267..9335))
                     /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     9098..9117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9336..9440)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9519..9538)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
       61 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      121 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      181 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      241 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      301 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      361 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      421 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      481 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      541 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      601 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      661 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
      721 atccgctaga gatccgcggc cgctaatacg actcactata gggagagccg ccaccatgaa
      781 acggacagcc gacggaagcg agttcgagtc accaaagaag aagcggaaag tctctgaggt
      841 ggagttttcc cacgagtact ggatgagaca tgccctgacc ctggccaaga gggcacgcga
      901 tgagaggaag gcccctgtgg gagccgtgct ggtgctgaac aatagagtga tcggcgaggg
      961 ctggaacaga gccatcggcc tgcacgaccc aacagcccat gccgaaatta tcgccctgag
     1021 acagggcggc ctggtcatgc agaactacag actgattgac gccaccctgt acgtgacatt
     1081 cgagccttgc gtgatgtgcg ccggcgccat gatcaactct aggatcggcc gcgtggtgtt
     1141 tggcgtgagg aacagcaaaa gaggcgccgc aggctccctg atgaacgtgc tgaactaccc
     1201 cggcatgaat caccgcgtcg aaattaccga gggaatcctg gcagatgaat gtgccgccct
     1261 gctgtgcgat ttctaccgga tgcctagaca ggtgttcaat gctcagaaga aggcccagag
     1321 ctccatcaac tccggaggat ctagcggagg ctcctctggc tctgagacac ctggcacaag
     1381 cgagagcgca acacctgaaa gcagcggggg cagcagcggg gggtcagaca agaagtacag
     1441 catcggcctg gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     1501 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     1561 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     1621 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     1681 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     1741 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     1801 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     1861 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     1921 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     1981 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     2041 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     2101 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     2161 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     2221 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     2281 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     2341 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     2401 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     2461 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     2521 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     2581 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     2641 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     2701 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     2761 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     2821 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     2881 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     2941 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     3001 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     3061 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     3121 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     3181 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     3241 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     3301 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     3361 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     3421 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     3481 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     3541 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     3601 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     3661 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     3721 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     3781 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     3841 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     3901 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     3961 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     4021 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     4081 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     4141 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     4201 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     4261 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     4321 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     4381 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     4441 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     4501 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     4561 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     4621 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     4681 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     4741 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     4801 ggaccctaag aagtacggcg gcttcctgtg gcccaccgtg gcctattctg tgctggtggt
     4861 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     4921 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     4981 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     5041 ggaaaacggc cggaagagaa tgctggcctc tgccagatac ctgcagaagg gaaacgaact
     5101 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     5161 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     5221 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     5281 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     5341 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     5401 ctttgacacc accatcgacc ggaaggagta caagagcacc aaagaggtgc tggacgccac
     5461 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     5521 aggtgacagc ggcgggagcg gcgggagcgg ggggagcact aatctgagcg acatcattga
     5581 gaaggagact gggaaacagc tggtcattca ggagtccatc ctgatgctgc ctgaggaggt
     5641 ggaggaagtg atcggcaaca agccagagtc tgacatcctg gtgcacaccg cctacgacga
     5701 gtccacagat gagaatgtga tgctgctgac ctctgacgcc cccgagtata agccttgggc
     5761 cctggtcatc caggattcta acggcgagaa taagatcaag atgctgagcg gaggatccgg
     5821 aggatctgga ggcagcacca acctgtctga catcatcgag aaggagacag gcaagcagct
     5881 ggtcatccag gagagcatcc tgatgctgcc cgaagaagtc gaagaagtga tcggaaacaa
     5941 gcctgagagc gatatcctgg tccataccgc ctacgacgag agtaccgacg aaaatgtgat
     6001 gctgctgaca tccgacgccc cagagtataa gccctgggct ctggtcatcc aggattccaa
     6061 cggagagaac aaaatcaaaa tgctgtctgg cggctcaaaa agaaccgccg acggcagcga
     6121 attcgagccc aagaagaaga ggaaagtcgg aagcggagct actaacttca gcctgctgaa
     6181 gcaggctgga gacgtggagg agaaccctgg acctatggtg agcaagggcg aggagctgtt
     6241 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     6301 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     6361 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctatggagt
     6421 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     6481 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     6541 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     6601 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     6661 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     6721 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     6781 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     6841 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     6901 gatcactctc ggcatggacg agctgtacaa gtctggtggt tctcccaaga agaagaggaa
     6961 agtctaaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
     7021 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     7081 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     7141 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     7201 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     7261 accagctggg gctcgatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     7321 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7381 ataaagtgta aagcctaggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7441 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7501 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7561 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7621 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     8101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     8161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     8221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     8281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacact
     8341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     8401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8461 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8521 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     8581 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8641 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8701 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8761 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8821 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8881 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8941 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     9001 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     9061 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     9121 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     9181 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     9241 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     9301 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     9361 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9421 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     9481 ggagatcgat ctcccgatcc cctagggtcg actctcagta caatctgctc tgatgccgca
     9541 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta g
//
