LOCUS       sequence-447648-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with IRAVQL 
            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(IRAVQL)-P2A-EGFP (RTW3820)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(89..106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(260..848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(340..359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1019..1810,1811..1879))
                     /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     1642..1661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1880..1984)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2063..2082)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2254..2633
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2634..2837
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2788..2808
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2879..2898
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2879..2897
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2905..2914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2911..7014
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFIRPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAAVLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYLSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7069..7089
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7102..7167
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7186..7242
                     /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(7243..7245,7246..7248,7249..7962)
                     /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(7243..7245,7246..7248,7249..7959)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7549..7568)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7896..7917
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7968..7985
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8008..8025)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8014..8238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8309..8325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8309..8325)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8322..8344)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8333..8349
                     /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(8357..8363,8364..8381,8382..8387))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8402..8423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
       61 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
      121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      241 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      421 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      661 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      781 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      841 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      901 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      961 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1021 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1081 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1141 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1201 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1261 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1321 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1381 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1441 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1501 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1561 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1621 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1681 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1741 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1801 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1861 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1921 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1981 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
     2041 atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     2101 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     2161 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     2221 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     2281 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     2341 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2401 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     2461 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     2521 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     2581 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     2641 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     2701 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     2761 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     2821 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagagatcc gcggccgcta
     2881 atacgactca ctatagggag agccgccacc atggacaaga agtacagcat cggcctggac
     2941 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     3001 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     3061 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     3121 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     3181 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     3241 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     3301 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     3361 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     3421 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     3481 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     3541 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     3601 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     3661 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     3721 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     3781 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     3841 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     3901 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     3961 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     4021 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     4081 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     4141 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     4201 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     4261 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     4321 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     4381 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     4441 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     4501 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     4561 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     4621 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     4681 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     4741 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     4801 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     4861 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     4921 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     4981 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     5041 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     5101 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     5161 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     5221 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     5281 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     5341 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     5401 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     5461 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
     5521 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     5581 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     5641 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     5701 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     5761 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     5821 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     5881 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     5941 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     6001 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     6061 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     6121 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     6181 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     6241 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     6301 tacggcggct tcatcagacc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     6361 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     6421 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     6481 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     6541 aagagaatgc tggcctctgc cgccgtgctg cagaagggaa acgaactggc cctgccctcc
     6601 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     6661 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     6721 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     6781 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     6841 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
     6901 atcgaccgga agcagtacct gagcaccaaa gaggtgctgg acgccaccct gatccaccag
     6961 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacggatcc
     7021 ggcggaggcg gaagcgggaa aagaaccgcc gacggcagcg aattcgagcc caagaagaag
     7081 aggaaagtct cgagcggagg cgactacaaa gaccatgacg gtgattataa agatcatgac
     7141 atcgattaca aggatgacga tgacaaggga ggcggtggaa gcggcgcaac aaacttctct
     7201 ctgctgaaac aagccggaga tgtcgaagag aatcctggac cgatggtgag caagggcgag
     7261 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     7321 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     7381 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     7441 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     7501 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     7561 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     7621 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     7681 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     7741 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     7801 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     7861 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     7921 gccgccggga tcactctcgg catggacgag ctgtacaagt gaccggtcat catcaccatc
     7981 accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     8041 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     8101 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     8161 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     8221 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga
     8281 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     8341 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct
     8401 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc c
//
