LOCUS       sequence-447653-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with KWKYES 
            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 8451)
  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 8451)
  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-SpCas9(KWKYES)-P2A-EGFP (RAS2900)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(69..86)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(240..828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(320..339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(999..1790,1791..1859))
                     /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     1622..1641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1860..1964)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2043..2062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2234..2613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2614..2817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2768..2788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2859..2878
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2859..2877
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2885..2894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2891..6994
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFKWPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKYLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYSSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7049..7069
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7082..7147
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7166..7222
                     /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(7223..7225,7226..7228,7229..7942)
                     /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(7223..7225,7226..7228,7229..7939)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7529..7548)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7876..7897
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7948..7965
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7988..8005)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7994..8218
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8289..8305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8289..8305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8302..8324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8313..8329
                     /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(8337..8343,8344..8361,8362..8367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8382..8403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
       61 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
      121 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      181 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      241 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      301 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      361 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      421 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      481 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      541 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      601 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      661 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      721 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      781 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      841 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      901 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
      961 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     1021 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     1081 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     1141 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1201 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1261 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     1321 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     1381 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     1441 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     1501 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     1561 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     1621 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     1681 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     1741 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     1801 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     1861 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1921 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     1981 gtgccacctg acgtcgacgg atcgggagat cgatctcccg atcccctagg gtcgactctc
     2041 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     2101 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     2161 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     2221 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     2281 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     2341 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     2401 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     2461 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     2521 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     2581 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     2641 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     2701 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     2761 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     2821 ttagtgaacc gtcagatccg ctagagatcc gcggccgcta atacgactca ctatagggag
     2881 agccgccacc atggacaaga agtacagcat cggcctggac atcggcacca actctgtggg
     2941 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     3001 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     3061 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     3121 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     3181 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     3241 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     3301 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     3361 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     3421 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     3481 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     3541 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     3601 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     3661 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     3721 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     3781 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     3841 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     3901 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     3961 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     4021 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     4081 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     4141 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     4201 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     4261 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     4321 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     4381 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     4441 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     4501 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     4561 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     4621 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     4681 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     4741 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     4801 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     4861 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     4921 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     4981 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     5041 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     5101 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     5161 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     5221 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     5281 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     5341 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     5401 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     5461 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     5521 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     5581 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     5641 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     5701 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     5761 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     5821 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     5881 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     5941 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     6001 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     6061 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     6121 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     6181 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     6241 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcaagtggcc
     6301 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     6361 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     6421 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     6481 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     6541 caagtacctg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     6601 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     6661 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     6721 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     6781 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     6841 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga aggagtacag
     6901 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     6961 gacacggatc gacctgtctc agctgggagg cgacggatcc ggcggaggcg gaagcgggaa
     7021 aagaaccgcc gacggcagcg aattcgagcc caagaagaag aggaaagtct cgagcggagg
     7081 cgactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga
     7141 tgacaaggga ggcggtggaa gcggcgcaac aaacttctct ctgctgaaac aagccggaga
     7201 tgtcgaagag aatcctggac cgatggtgag caagggcgag gagctgttca ccggggtggt
     7261 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     7321 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     7381 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     7441 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     7501 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     7561 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     7621 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     7681 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     7741 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     7801 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     7861 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     7921 catggacgag ctgtacaagt gaccggtcat catcaccatc accattgagt ttaaacccgc
     7981 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     8041 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     8101 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     8161 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     8221 tctgaggcgg aaagaaccag ctggggctcg ataccgtcga cctctagcta gagcttggcg
     8281 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     8341 atacgagccg gaagcataaa gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca
     8401 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg c
//
